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Sample records for benzopyrene-diol-epoxide bpde-induced bax

  1. Inhibition of benzopyrene-diol-epoxide (BPDE)-induced bax and caspase-9 by cadmium: Role of mitogen activated protein kinase

    Cadmium, a major metal constituent of tobacco smoke, elicits synergistic enhancement of cell transformation when combined with benzo[a]pyrene (BP) or other polynuclear aromatic hydrocarbons (PAHs). The mechanism underlying this synergism is not clearly understood. Present study demonstrates that (+/-)-anti-benzo[a]pyrene-7,8-diol-9,10-epoxide (BPDE), an ultimate carcinogen of BP, induces apoptosis in human leukemic HL-60 cells and others, and cadmium at non-cytotoxic concentration inhibits BPDE-induced apoptosis. We observed that BPDE treatment also activates all three MAP kinases e.g. ERK1/2, p38 and JNK in HL-60 cells, and inhibition of BPDE-induced apoptosis by cadmium is associated with down-regulation of pro-apoptotic bax induction/caspase-9 activation and up-regulation of ERK phosphorylation, whereas p38 MAP kinase and c-Jun phosphorylation (indicative of JNK activation) remain unaffected. Inhibition of ERKs by prior treatment of cells with 10 μM U0126 relieves cadmium-mediated inhibition of apoptosis/bax induction/caspase-9 activation. Our results suggest that cadmium inhibits BPDE-induced apoptosis by modulating apoptotic signaling through up-regulation of ERK, which is known to promote cell survival

  2. Correlation between base-excision repair gene polymorphisms and levels of in-vitro BPDE-induced DNA adducts in cultured peripheral blood lymphocytes.

    Hongping Yu

    Full Text Available In vitro benzo[a]pyrene diol epoxide (BPDE-induced DNA adducts in cultured peripheral lymphocytes have been shown to be a phenotypic biomarker of individual's DNA repair phenotype that is associated with cancer risk. In this study, we explored associations between genotypes of base-excision repair genes (PARP1 Val762Ala, APEX1 Asp148Glu, and XRCC1 Arg399Gln and in vitro BPDE-induced DNA adducts in cultured peripheral blood lymphocytes in 706 cancer-free non-Hispanic white subjects. We found that levels of BPDE-induced DNA adducts were significantly higher in ever smokers than in never smokers and that individuals with the Glu variant genotypes (i.e., Asp/Glu and Glu/Glu exhibited lower levels of BPDE-induced DNA adducts than did individuals with the common Asp/Asp homozygous genotype (median RAL levels: 32.0 for Asp/Asp, 27.0 for Asp/Glu, and 17.0 for Glu/Glu, respectively; P(trend = 0.030. Further stratified analysis showed that compared with individuals with the common APEX1-148 homozygous Asp/Asp genotype, individuals with the APEX1-148Asp/Glu genotype or the Glu/Glu genotype had a lower risk of having higher-level adducts (adjusted OR = 0.60, 95% CI: 0.36-0.98 and adjusted OR = 0.47, 95% CI: 0.26-0.86, respectively; P(trend = 0.012 among smokers. Such an effect was not observed in non-smokers. However, there was no significant interaction between the APEX1 Asp148Glu polymorphism and smoking exposure in this study population (P = 0.512. Additional genotype-phenotype analysis found that the APEX1-148Glu allele had significantly increased expression of APEX1 mRNA in 270 Epstein-Barr virus-transformed lymphoblastoid cell lines, which is likely associated with more active repair activity. Our findings suggest that the functional APEX1-148Glu allele is associated with reduced risk of having high levels of BPDE-induced DNA adducts mediated with high levels of mRNA expression.

  3. BAX supports the mitochondrial network, promoting bioenergetics in nonapoptotic cells

    Boohaker, Rebecca J.; Zhang, Ge; Carlson, Adina Loosley; Nemec, Kathleen N.

    2011-01-01

    The dual functionality of the tumor suppressor BAX is implied by the nonapoptotic functions of other members of the BCL-2 family. To explore this, mitochondrial metabolism was examined in BAX-deficient HCT-116 cells as well as primary hepatocytes from BAX-deficient mice. Although mitochondrial density and mitochondrial DNA content were the same in BAX-containing and BAX-deficient cells, MitoTracker staining patterns differed, suggesting the existence of BAX-dependent functional differences in mitochondrial physiology. Oxygen consumption and cellular ATP levels were reduced in BAX-deficient cells, while glycolysis was increased. These results suggested that cells lacking BAX have a deficiency in the ability to generate ATP through cellular respiration. This conclusion was supported by detection of reduced citrate synthase activity in BAX-deficient cells. In nonapoptotic cells, a portion of BAX associated with mitochondria and a sequestered, protease-resistant form was detected. Inhibition of BAX with small interfering RNAs reduced intracellular ATP content in BAX-containing cells. Expression of either full-length or COOH-terminal-truncated BAX in BAX-deficient cells rescued ATP synthesis and oxygen consumption and reduced glycolytic activity, suggesting that this metabolic function of BAX was not dependent upon its COOH-terminal helix. Expression of BCL-2 in BAX-containing cells resulted in a subsequent loss of ATP measured, implying that, even under nonapoptotic conditions, an antagonistic interaction exists between the two proteins. These findings infer that a basal amount of BAX is necessary to maintain energy production via aerobic respiration. PMID:21289292

  4. Ernest Belfort Bax: Marxist, Idealist, Positivist

    Bevir, Mark

    1993-01-01

    Bax was the leading philosopher of the socialist revival in Britain during the 1880s. He saw Marxism as an economic and historical science that lacked a philosophical and ethical basis. Consequently, he tried to justify the Marxian dialectic by using a philosophy indebted to German idealism to show that the dialectic was a fact about reality itself, and he also tried to provide an ethical defence of Marxism in terms of a positivist ethic enshrining the goals of the French Revolution. Such ...

  5. Bax-induced cell death in Candida albicans.

    De Smet, Kris; Eberhardt, Ines; Reekmans, Rieka; Contreras, Roland

    2004-12-01

    Bax is a pro-apoptotic member of the Bcl-2 family of proteins involved in the regulation of genetically programmed cell death in mammalian cells. It has been shown that heterologous expression of Bax in several yeast species, such as Saccharomyces cerevisiae, Schizosaccharomyces pombe and Pichia pastoris, also induces cell death. In this study we investigated the effects of Bax expression in the pathogenic yeast Candida albicans. Cell death inducing expression of Bax required a synthetic BAX gene that was codon-optimized for expression in Candida albicans. Expression of this BAX gene resulted in growth inhibition and cell death. By fusing Bax with the yeast enhanced green fluorescent protein of Aequoria victoria, the cell death-inducing effect of Bax was increased due to reduced proteolytic degradation of Bax. Using this fusion protein we showed that, upon expression in C. albicans, Bax co-localizes with the mitochondria. Furthermore, we showed for the first time that expression of Bax in yeast causes the mitochondria, which are normally distributed throughout the cell, to cluster in the perinuclear region. PMID:15565645

  6. Small molecules reveal an alternative mechanism of Bax activation.

    Brahmbhatt, Hetal; Uehling, David; Al-Awar, Rima; Leber, Brian; Andrews, David

    2016-04-15

    The pro-apoptotic protein Bax commits a cell to death by permeabilizing the mitochondrial outer membrane (MOM). To obtain small-molecule probes for elucidating the molecular mechanism(s) of Bax activation, we screened for compounds that induced Bax-mediated liposome permeabilization. We identified five structurally different small molecules that promoted both Bax targeting to and oligomerization at membranes. All five compounds initiated Bax oligomerization in the absence of membranes by a mechanism unlike Bax activation by Bcl-2 homology 3 domain (BH3) proteins. Some of the compounds induced Bax/Bak-dependent apoptosis in cells. Activation of Bax by the most active compound was poorly inhibited by the anti-apoptotic protein Bcl-XL and requires a cysteine residue at position 126 of Bax that is not required for activation by BH3 proteins. Our results reveal a novel pathway for Bax activation independent of pro-apoptotic BH3 proteins that may have important implications for the regulation of Bax activity in cells. PMID:26916338

  7. PUMA and Bax-induced Autophagy Contributes to Apoptosis

    Yee, Karen S.; Wilkinson, Simon; James, John; Ryan, Kevin M.; Vousden, Karen H.

    2009-01-01

    The p53-inducible BH3-only protein PUMA is a key mediator of p53-dependent apoptosis, and PUMA has been shown to function by activating Bax and mitochondrial outer membrane permeabilization. In this study we describe an ability of PUMA to induce autophagy that leads to the selective removal of mitochondria. This function of PUMA depends on Bax/Bak and can be reproduced by overexpression of Bax. The induction of autophagy coincides with cytochrome c release, and taken together the results sugg...

  8. PUMA- and Bax-induced autophagy contributes to apoptosis

    Yee, K S; Wilkinson, S; James, J; Ryan, K M; Vousden, K H

    2009-01-01

    The p53-inducible BH3-only protein PUMA is a key mediator of p53-dependent apoptosis, and PUMA has been shown to function by activating Bax and mitochondrial outer membrane permeabilization. In this study, we describe an ability of PUMA to induce autophagy that leads to the selective removal of mitochondria. This function of PUMA depends on Bax/Bak and can be reproduced by overexpression of Bax. The induction of autophagy coincides with cytochrome c release, and taken together the results sug...

  9. The apoptotic response in HCT116BAX-/- cancer cells becomes rapidly saturated with increasing expression of a GFP-BAX fusion protein

    Many chemotherapeutic agents promote tumor cell death by activating the intrinsic pathway of apoptosis. Intrinsic apoptosis involves permeabilization of the mitochondrial outer membrane and the release of cytochrome c, a process that is controlled by proteins of the BCL2 gene family. Chemoresistance is often associated with abnormalities in concentrations of BCL2 family proteins. Although stoichiometirc interactions between anti-apoptotic and BH3-only BCL2 family proteins have been well documented as affecting cell death, the association between changes in BAX concentration and intrinsic apoptosis are poorly understood. Exogenous GFP-murine Bax fusion constructs were transfected into BAX-deficient HCT116 cells. To titrate the expression of the fusion protein, GFP-BAX was cloned into a tetracycline sensitive expression cassette and cotransfected with a plasmid expressing the rtTA transcription factor into HCT116BAX-/- cells. Linear expression of the fusion gene was induced with doxycycline and monitored by quantitative PCR and immunoblotting. Cell death was assayed by DAPI staining cells after exposure to indomethacin, and scoring nuclei for condensed chromatin and fragmented nuclei. HCT116BAX-/- cells were resistant to indomethacin, but susceptibility could be recovered in cells expressing a GFP-BAX fusion protein. Titration of GFP-BAX expression revealed that the concentration of BAX required to induce a saturating apoptosis response from baseline, was rapidly achieved. Increased levels of GFP-BAX were unable to stimulate higher levels of cell death. Examination of GFP-BAX distribution before and after indomethacin treatment indicated that BAX protein did not form aggregates when present at sub-lethal concentrations. Within the limitations of this experimental system, BAX-dependent apoptosis in HCT116 cells exhibits an all-or-none response depending on the level of BAX protein present. The lack of BAX aggregation at sub-saturation levels suggests that the

  10. Spatial and temporal changes in Bax subcellular localization during NPe6-PDT-induced apoptosis

    Liu, Lei; Xing, Da; Chen, Wei R.; Wan, Qingling; Zhou, Feifan

    2008-02-01

    Photodynamic therapy (PDT) employing photosensiter N-aspartyl chlorin e6 (NPe6) can induce lysosome disruption and initiate the intrinsic apoptotic pathway. Bax, a member of the Bcl-2 family of proteins, is an essential regulator of apoptosis. Bax is normally found in the cytosol of healthy cells, and translocates to mitochondria in response to many apoptotic stimuli. In this study, using real-time single-cell analysis, we have investigated the kinetics of Bax distribution during NPe6-induced apoptosis in ASTC-a-1 cells. In order to monitor Bax subcellular distribution, cells were stained with GFP-Bax and Mito Tracker Red. The results show that Bax redistribution occurred at about 170 min after treated with NPe6-PDT, and then sequestered into clusters associated with the mitochondira within 30 min. Our data clearly showed the spatial and temporal changes in Bax distribution in living cells during NPe6-induced apoptosis.

  11. Overexpression of Bax induces apoptosis and enhances drug sensitivity of hepatocellular cancer-9204 cells

    Jian-Yong Zheng; Guang-Shun Yang; Wei-Zhong Wang; Jiang Li; Kai-Zong Li; Wen-Xian Guan; Wen-Liang Wang

    2005-01-01

    AIM: To investigate the role of overexpression of Bax in apoptotic pathways and the response of human hepatocellular cancer (HCC)-9204 cells to cell death induced by adriamycin.METHODS: The whole length of Bax cDNA was transfectrd into human HCC-9204 cells by the method of lipofectamine transfection. An inducible MT-Ⅱ regulatory system was constructed, which allowed controlled expression of protein upon addition of ZnSO4 (100 μmol/L) as an external inducer. Stable transfecting inducible expression vector containing Bax gene was performed. Expression of Bax in protein was analyzed by immunohistochemistry and Western blotting. TUNEL and flow cytometry were used to assess the effect of Bax on apoptosis. Colony assay and tetrazolium blue (MTT) assay were used to evaluate the difference in drug sensitivity of HCC-9204 cells after Bax-transfection.RESULTS: Immunohistochemistry and Western blotting demonstrated that the expression of Bax protein markedly increased in Bax-transfected cells 4 h after the addition cytoplasm and perinuclear region of HCC-9404 cells, and there was ectopic expression in cells with marked condensation of chromatin and cytoplasm (apoptotic cells). Apoptotic index significantly increased in Bax-transfected HCC-9204/Bax cells (3.6 vs 27.2, 4.2 vs 32.3, P<0.05).Flow cytometry analysis showed a significant sub-G1 peak and apoptosis in 15.4% HCC-9204/Bax cells 24 h after treatment. Furthermore, colony survival rate decreased from 66% (HCC-9204/pMD) to 45% (HCC-9204/Bax) 2 dafter ADR withdrawal. MTT assay result showed that the effects of Bax on cell viability following ADR exposure were significant as compared to the vehicle-transfected HCC-9204/pMD cells (21% vs 44%, P<0.01).CONCLUSION: Overexpression of Bax not only induces apoptosis, but also sensitizes HCC-9204 cells to cell death induced by adriamycin.

  12. Clinicopathological significance of Bcl-2 and Bax protein expression in human pancreatic cancer

    Ming Dong; Jian-Ping Zhou; Hao Zhang; Ke-Jian Guo; Yu-Lin Tian; Yu-Ting Dong

    2005-01-01

    AIM: To assess the clinicopathological significance of the expression of the apoptosis-inhibitory Bcl-2 protein (pBcl-2) and the apoptosis-promoting Bax protein (pBax) in human invasive ductal carcinomas (IDCs) of the pancreas. METHODS: Fifty-nine surgical specimens of IDCs of the pancreas were stained immunohistochemically to detectpBcl-2 and pBax expressions whose correlation to tumor classification, staging, and prognosis was analyzed by univariate and multivariate analyses. RESULTS: The expression of pBcl-2 and pBax was detected in 21 of 59 (35.6%) and in 29 of 59 (49.2%) patients with IDCs of the pancreas, respectively. Neither pBcl-2 nor pBax alone was correlated to TNM staging and differentiation degree of IDCs of the pancreas according to univariate analysis. By Mantel-Cox test, the median survival time after surgery for pBcl-2(+) and pBcl-2(-) groups were 14.3 and 7.3 mo, respectively (χ2= 9.357, P = 0.002) and that for pBax(+) and pBax(-) groups were 12.9 and 10.2 mo, respectively (χ2= 0.285, P>0.05).Contingency coefficient between pBd-2 and pBax expression was 0.298, indicating that there was correlation between them (χ2= 5.74, P<0.05). The median survival time after surgery for pBd-2(+)pBax(+) and pBcl-2(+)pBax(-) groups were 14.3 and 14.1 mo, respectively, and that for pBcl-2 (-)pBax(+) and pBcl-2(-)pBax(-) groups were 5.9 and 9.9 mo, respectively. There was a significant difference between pBcl-2(+)pBax(+) and pBcl-2(-)pBax(+) (χ2 = 5.06,P<0.05), such was the case for pBcl-2(+)pBax(+) andpBcl-2(-)pBax(-) (χ2= 7.18, P<0.01). Cox proportional hazards model for multivariate analysis was applied, indicating that pBcl-2, TNM staging, age and pBax were high risk factors of post-surgical survival time. CONCLUSION: Both pBcl-2 and pBax have high expression in IDCs of the pancreas, indicating that co-expression of pBcl-2 and pBax is a good indicator of favorable prognosis in IDCs of the pancreas.

  13. Bax function in the absence of mitochondria in the primitive protozoan Giardia lamblia.

    Adrian B Hehl

    Full Text Available Bax-induced permeabilization of the mitochondrial outer membrane and release of cytochrome c are key events in apoptosis. Although Bax can compromise mitochondria in primitive unicellular organisms that lack a classical apoptotic machinery, it is still unclear if Bax alone is sufficient for this, or whether additional mitochondrial components are required. The protozoan parasite Giardia lamblia is one of the earliest branching eukaryotes and harbors highly degenerated mitochondrial remnant organelles (mitosomes that lack a genome. Here we tested whether human Bax expressed in Giardia can be used to ablate mitosomes. We demonstrate that these organelles are neither targeted, nor compromised, by Bax. However, specialized compartments of the regulated secretory pathway are completely ablated by Bax. As a consequence, maturing cyst wall proteins that are sorted into these organelles are released into the cytoplasm, causing a developmental arrest and cell death. Interestingly, this ectopic cargo release is dependent on the carboxy-terminal 22 amino acids of Bax, and can be prevented by the Bax-inhibiting peptide Ku70. A C-terminally truncated Bax variant still localizes to secretory organelles, but is unable to permeabilize these membranes, uncoupling membrane targeting and cargo release. Even though mitosomes are too diverged to be recognized by Bax, off-target membrane permeabilization appears to be conserved and leads to cell death completely independently of mitochondria.

  14. Bax function in the absence of mitochondria in the primitive protozoan Giardia lamblia.

    Hehl, Adrian B; Regos, Attila; Schraner, Elisabeth; Schneider, André

    2007-01-01

    Bax-induced permeabilization of the mitochondrial outer membrane and release of cytochrome c are key events in apoptosis. Although Bax can compromise mitochondria in primitive unicellular organisms that lack a classical apoptotic machinery, it is still unclear if Bax alone is sufficient for this, or whether additional mitochondrial components are required. The protozoan parasite Giardia lamblia is one of the earliest branching eukaryotes and harbors highly degenerated mitochondrial remnant organelles (mitosomes) that lack a genome. Here we tested whether human Bax expressed in Giardia can be used to ablate mitosomes. We demonstrate that these organelles are neither targeted, nor compromised, by Bax. However, specialized compartments of the regulated secretory pathway are completely ablated by Bax. As a consequence, maturing cyst wall proteins that are sorted into these organelles are released into the cytoplasm, causing a developmental arrest and cell death. Interestingly, this ectopic cargo release is dependent on the carboxy-terminal 22 amino acids of Bax, and can be prevented by the Bax-inhibiting peptide Ku70. A C-terminally truncated Bax variant still localizes to secretory organelles, but is unable to permeabilize these membranes, uncoupling membrane targeting and cargo release. Even though mitosomes are too diverged to be recognized by Bax, off-target membrane permeabilization appears to be conserved and leads to cell death completely independently of mitochondria. PMID:17534438

  15. Expression of Bcl-2 and Bax in extrahepatic biliary tract carcinoma and dysplasia

    Sheng-Mian Li; Shu-Kun Yao; Nobuyoshi Yamamura; Toshitsugu Nakamura

    2003-01-01

    AIM: To compare the difference of expression of Bcl-2 and Bax in extrahepatic biliary tract carcinoma and dysplasia, and to analyze the role of Bcl-2 and Bax proteins in the progression from dysplasia to carcinoma and to evaluate the correlation of Bcl-2/Bax protein expression with the biological behaviors.METHODS: Expressions of Bcl-2 and Bax were examined immunohistochemically in 27 cases of extrahepatic biliary tract carcinomas (bile duct carcinoma: n=21, carcinoma of ampulla of Vater: n=6), and 10 cases of atypical dysplasia.Five cases of normal biliary epithelial tissues were used as controls. A semiquantitative scoring system was used to assess the Bcl-2 and Bax reactivity.RESULTS: The expression of Bd-2 was observed in 10 out of 27 (37.0 %) invasive carcinomas, 1 out of 10 clysplasias, none out of 5 normal epithelial tissues. Bax expression rate was 74.1% (20/27) in invasive carcinoma, 30 % (3/10) in dysplasia,and 40 % (2/5) in normal biliary epithelium. Bcl-2 and Bax activities were more intense in carcinoma than in dysplasia,with no significant difference in Bcl-2 expression (P=0.1:10),and significant difference in Bax expression (P=0.038). Level of Bax expression was higher in invasive carcinoma than in dysplasia and normal tissue (P=0.012). Bcl-2 expression was correlated to Bax expression (P=0.0059). However, Bcl-2/Bax expression had no correlation with histological subtype,grade of differentiation, or level of invasion.CONCLUSION: Increased Bcl-2/Bax expression from dysplasia to invasive tumors supports the view that this is the usual route for the development of extrahepatic biliary tract carcinoma. Bcl-2/Bax may be involved, at least in part,in the apoptotic activity in extrahepatic biliary carcinoma.

  16. Evidence that inhibition of BAX activation by BCL-2 involves its tight and preferential interaction with the BH3 domain of BAX

    Bonsu Ku; Chengyu Liang; Jae U Jung; Byung-Ha Oh

    2011-01-01

    Interactions between the BCL-2 family proteins determine the cell's fate to live or die. How they interact with each other to regulate apoptosis remains as an unsettled central issue. So far, the antiapoptotic Bc1-2 proteins are thought to interact with BAX weakly, but the physiological significance of this interaction has been vague.Herein, we show that recombinant BCL-2 and BCL-w interact potently with a BCL-2 homology (BH) 3 domain-containing peptide derived from BAX, exhibiting the dissociation constants of 15 and 23 nM, respectively. To clarify the basis for this strong interaction, we determined the three-dimensional structure of a complex of BCL-2 with a BAX peptide spanning its BH3 domain. It revealed that their interactions extended beyond the canonical BH3 domain and involved three nonconserved charged residues of BAX. A novel BAX variant, containing the alanine substitution of these three residues, had greatly impaired affinity for BCL-2 and BCL-w, hut was otherwise indistinguishable from wild-type BAX. Critically, the apoptotic activity of the BAX variant could not be restrained by BCL-2 and BCL-w, pointing that the observed tight interactions are critical for regulating BAX activation. We also comprehensively quantified the binding affinities between the three BCL-2 subfamily proteins. Collectively, the data show that due to the high affinity of BAX for BCL-2, BCL-w and A1, and of BAK for BCL-XL, MCL-1 and A1, only a subset of BH3-only proteins, commonly including BIM, BID and PUMA, could he expected to free BAX or BAK from the antiapoptotic BCL-2 proteins to elicit apoptosis.

  17. The oxidized phospholipid PazePC promotes permeabilization of mitochondrial membranes by Bax.

    Lidman, Martin; Pokorná, Šárka; Dingeldein, Artur P G; Sparrman, Tobias; Wallgren, Marcus; Šachl, Radek; Hof, Martin; Gröbner, Gerhard

    2016-06-01

    Mitochondria play a crucial role in programmed cell death via the intrinsic apoptotic pathway, which is tightly regulated by the B-cell CLL/lymphoma-2 (Bcl-2) protein family. Intracellular oxidative stress causes the translocation of Bax, a pro-apoptotic family member, to the mitochondrial outer membrane (MOM) where it induces membrane permeabilization. Oxidized phospholipids (OxPls) generated in the MOM during oxidative stress directly affect the onset and progression of mitochondria-mediated apoptosis. Here we use MOM-mimicking lipid vesicles doped with varying concentrations of 1-palmitoyl-2-azelaoyl-sn-glycero-3-phosphocholine (PazePC), an OxPl species known to significantly enhance Bax-membrane association, to investigate three key aspects of Bax's action at the MOM: 1) induction of Bax pores in membranes without additional mediator proteins, 2) existence of a threshold OxPl concentration required for Bax-membrane action and 3) mechanism by which PazePC disturbs membrane organization to facilitate Bax penetration. Fluorescence leakage studies revealed that Bax-induced leakage, especially its rate, increased with the vesicles' PazePC content without any detectable threshold neither for OxPl nor Bax. Moreover, the leakage rate correlated with the Bax to lipid ratio and the PazePC content. Solid state NMR studies and calorimetric experiments on the lipid vesicles confirmed that OxPl incorporation disrupted the membrane's organization, enabling Bax to penetrate into the membrane. In addition, 15N cross polarization (CP) and insensitive nuclei enhanced by polarization transfer (INEPT) MAS NMR experiments using uniformly (15)N-labeled Bax revealed dynamically restricted helical segments of Bax embedded in the membrane, while highly flexible protein segments were located outside or at the membrane surface. PMID:26947183

  18. Nonredundant Role of Bax and Bak in Bid-Mediated Apoptosis

    Cartron, Pierre-François; Juin, Philippe; Oliver, Lisa; Martin, Stéphane; Meflah, Khaled; Vallette, François M.

    2003-01-01

    Animal models suggest that Bax and Bak play an essential role in the implementation of apoptosis and as a result can hinder tumorigenesis. We analyzed the expression of these proteins in 50 human glioblastoma multiforme (GBM) tumors. We found that all the tumors expressed Bak, while three did not express Bax. In vitro, Bax-deficient GBM (BdGBM) exhibited an important resistance to various apoptogenic stimuli (e.g., UV, staurosporine, and doxorubicin) compared to the Bax-expressing GBM (BeGBM)...

  19. BimL involvement in Bax activation during UV irradiation-induced apoptosis

    Bax, a proapoptotic member of the Bcl-2 family, localizes largely in the cytoplasm but translocates to mitochondria and undergoes oligomerization to induce the release of apoptogenic factors in response to apoptotic stimuli. However, the molecular mechanism of Bax activation is not fully understood. We show here the role of BimL in Bax activation during UV irradiation-induced apoptosis. In this study, GFP-BimL plasmid was constructed. The dynamic interaction between BimL and Bax during UV irradiation-induced apoptosis was observed using fluorescence resonance energy transfer (FRET) technique. Our experimental results showed that BimL translocation to mitochondria occurred before Bax translocation, and that BimL activated Bax indirectly. Moreover, inhibition of c-Jun N-terminal protein kinase (JNK) activation blocked BimL translocation, delayed and attenuated Bax translocation and subsequent apoptosis. These results demonstrate that BimL is involved in UV irradiation-induced apoptosis by indirectly activating Bax

  20. Effect of Helicobacter pylori infection on Bax protein expression in patients with gastric precancerous lesions

    Hai-Feng Liu; Wei-Wen Liu; Guo-An Wang; Xiao-Chun Teng

    2005-01-01

    AIM: To investigate the effect of Helicobacter pylori (H pylori) infection on Bax protein expression, and explore the role of H pylori in gastric carcinogenesis.METHODS: H pylori was assessed by rapid urease test and Warthin-Starry method, and expression of Bax protein was examined immunohistochemically in 72 patients with pre-malignant lesions.RESULTS: Bax protein was differently expressed in intestinal metaplasia and gastric dysplasia, and showed 63.99% positivity. The positivity of Bax protein expression in H pylori-positive gastric precancerous lesions (72.3%) was significantly higher than that in H pylori-negative gastric precancerous lesions (48.0%, χ2 = 4.191, P<0.05).H pylori infection was well correlated with the expression of Bax protein in gastric precancerous lesions (r = 0.978,P<0.01). After eradication of H pylori, the positivity of Bax protein expression significantly decreased in H pylori-positive gastric precancerous lesions (χ2= 5.506,P<0.05). In the persisting H pylori-infected patients,the positivity of Bax protein expression was not changed.CONCLUSION: H pylori infection may be involved in the upregulation of Bax gene, which might be one of the mechanisms of H pylori infection-induced gastric epithelial cell apoptosis. H pylori might act as a tumor promoter in the genesis of gastric carcinoma and eradication of H pylori could inhibit gastric carcinogenesis.

  1. Involvement of nitric oxide signaling in mammalian Bax-induced terpenoid indole alkaloid production of Catharanthus roseus cells

    2007-01-01

    Bax, a mammalian pro-apoptotic member of the Bcl-2 family, has been demonstrated to be a potential regulatory factor for plant secondary metabolite biosynthesis recently. To investigate the molecular mechanism of Bax-induced secondary metabolite biosynthesis, we determined the contents of nitric oxide (NO) of the transgenic Catharanthus roseus cells overexpressing a mouse Bax protein and checked the effects of NO specific scavenger 2,4-carboxyphenyl-4,4,5,5-tetramethylimidazoline-1- oxyl-3-oxide (cPITO) on Bax-induced terpenoid indole alkaloid (TIA) production of the cells. The data showed that overexpression of the mouse Bax in C. roseus cells triggered NO generation of the cells. Treatment of cPITO not only inhibited the Bax-triggered NO burst but also suppressed the Bax-induced TIA production. The results indicated that the mouse Bax might activate the NO signaling in C. roseus cells and induce TIA production through the NO-dependent signal pathway in the cells. Furthermore, the activities of nitric oxide synthase (NOS) were significantly increased in the transgenic Bax cells as compared to those in the control cells, showing that the mouse Bax may induce NOS of C. roseus cells. Treatment of the transgenic Bax cells with NOS inhibitor PBITU blocked both Bax-induced NO generation and TIA production, which suggested that the mouse Bax might trigger NO generation and TIA production through NOS. However, the NOS-like activities and NO generation in the transgenic Bax cells did not match kinetically and the Bax-induced NOS-like activity was much later and lower than NO production. Moreover, the Bax-induced NO generation and TIA production were only partially inhibited by PBITU. Thus, our results suggested that the Bax-induced NO production and secondary metabolite biosynthesis in C. roseus cells was not entirely dependent on NOS or NOS-like enzymes.

  2. Bax/Bak activation in the absence of Bid, Bim, Puma, and p53.

    Zhang, J; Huang, K; O'Neill, K L; Pang, X; Luo, X

    2016-01-01

    How BH3-only proteins activate Bax/Bak, the two gateway proteins of the mitochondria-dependent apoptotic pathway, remains incompletely understood. Although all pro-apoptotic BH3-only proteins are known to bind/neutralize the anti-apoptotic Bcl-2 proteins, the three most potent ones, Bid (tBid), Bim, and Puma, possess an additional activity of directly activating Bax/Bak in vitro. This latter activity has been proposed to be responsible for triggering Bax/Bak activation following apoptotic stimulation. To test this hypothesis, we generated Bid(-/)(-)Bim(-/)(-)Puma(-/)(-) (TKO), TKO/Bax(-/)(-)/Bak(-/)(-) (PentaKO), and PentaKO/Mcl-1(-/-) (HexaKO) HCT116 cells through gene editing. Surprisingly, although the TKO cells were resistant to several apoptotic stimuli, robust apoptosis was induced upon the simultaneous inactivation of Bcl-xL and Mcl-1, two anti-apoptotic Bcl-2 proteins known to suppress Bax/Bak activation and activity. Importantly, such apoptotic activity was completely abolished in the PentaKO cells. In addition, ABT-737, a BH3 mimetic that inhibits Bcl-xL/Bcl-w/Bcl-2, induced Bax activation in HexaKO cells reconstituted with endogenous level of GFP-Bax. Further, by generating TKO/p53(-/-) (QKO) cells, we demonstrated that p53, a tumor suppressor postulated to directly activate Bax, is not required for Bid/Bim/Puma-independent Bax/Bak activation. Together, these results strongly suggest that the direct activation activities of Bid (tBid), Bim, Puma, and p53 are not essential for activating Bax/Bak once the anti-apoptotic Bcl-2 proteins are neutralized. PMID:27310874

  3. Molecular Basis for Membrane Pore Formation by Bax Protein Carboxyl Terminus

    Tatulian, Suren A.; Garg, Pranav; Nemec, Kathleen N.; Chen, Bo; Khaled, Annette R.

    2015-01-01

    Bax protein plays a key role in mitochondrial membrane permeabilization and cytochrome c release upon apoptosis. Our recent data have indicated that the 20-residue C-terminal peptide of Bax (BaxC-KK; VTIFVAGVL-TASLTIWKKMG), when expressed intracellularly, translocates to the mitochondria and exerts lethal effect on cancer cells. Moreover, the BaxC-KK peptide, as well as two mutants where the two lysines are replaced with glutamate (BaxC-EE) or leucine (BaxC-LL), have been shown to form relatively large pores in lipid membranes, composed of up to eight peptide molecules per pore. Here the pore structure is analyzed by polarized Fourier transform infrared, circular dichroism, and fluorescence experiments on the peptides reconstituted in phospholipid membranes. The peptides assume an α/β-type secondary structure within membranes. Both β-strands and α-helices are significantly (by 30–60 deg) tilted relative to the membrane normal. The tryptophan residue embeds into zwitterionic membranes at 8–9 Å from the membrane center. The membrane anionic charge causes a deeper insertion of tryptophan for BaxC-KK and BaxC-LL but not for BaxC-EE. Combined with the pore stoichiometry determined earlier, these structural constraints allow construction of a model of the pore where eight peptide molecules form an “α/β-ring” structure within the membrane. These results identify a strong membranotropic activity of Bax C-terminus and propose a new mechanism by which peptides can efficiently perforate cell membranes. Knowledge on the pore forming mechanism of the peptide may facilitate development of peptide-based therapies to kill cancer or other detrimental cells such as bacteria or fungi. PMID:23110300

  4. GSH depletion enhances adenoviral bax-induced apoptosis in lung cancer cells.

    Honda, Tsuyoshi; Coppola, Simona; Ghibelli, Lina; Cho, Song H; Kagawa, Shunsuke; Spurgers, Kevin B; Brisbay, Shawn M; Roth, Jack A; Meyn, Raymond E; Fang, Bingliang; McDonnell, Timothy J

    2004-04-01

    The utility of dominant acting proapoptotic molecules to induce cell death in cancer cells is being evaluated in preclinical studies and clinical trials. We recently developed a binary adenoviral expression system to enable the efficient gene transfer of Bax and other proapoptotic molecules. Using this system, overexpression of Bax protein in four non-small-cell lung cancer (NSCLC) cell lines, H1299, A549, H226 and H322, was evaluated. The H322 line exhibited significant resistance to Bax-induced cell death compared to the other cell lines. H322 cells had the highest level of glutathione (GSH). GSH levels were significantly decreased following buthionine sulfoximine treatment and this coincided with enhanced apoptosis induction by Ad-Bax in H322 cells. GSH depletion enhanced Bax protein translocation to mitochondrial membranes. These findings suggest that the redox status may be a determinant of Bax-mediated cell death and that manipulation of intracellular thiols may sensitize cells to apoptosis by facilitating Bax insertion into mitochondrial membranes. PMID:15002033

  5. Insights into the structural stability of Bax from molecular dynamics simulations at high temperatures

    Rosas-Trigueros, Jorge Luis; Correa-Basurto, José; Guadalupe Benítez-Cardoza, Claudia; Zamorano-Carrillo, Absalom

    2011-01-01

    Bax is a member of the Bcl-2 protein family that participates in mitochondrion-mediated apoptosis. In the early stages of the apoptotic pathway, this protein migrates from the cytosol to the outer mitochondrial membrane, where it is inserted and usually oligomerizes, making cytochrome c-compatible pores. Although several cellular and structural studies have been reported, a description of the stability of Bax at the molecular level remains elusive. This article reports molecular dynamics simulations of monomeric Bax at 300, 400, and 500 K, focusing on the most relevant structural changes and relating them to biological experimental results. Bax gradually loses its α-helices when it is submitted to high temperatures, yet it maintains its globular conformation. The resistance of Bax to adopt an extended conformation could be due to several interactions that were found to be responsible for maintaining the structural stability of this protein. Among these interactions, we found salt bridges, hydrophobic interactions, and hydrogen bonds. Remarkably, salt bridges were the most relevant to prevent the elongation of the structure. In addition, the analysis of our results suggests which conformational movements are implicated in the activation/oligomerization of Bax. This atomistic description might have important implications for understanding the functionality and stability of Bax in vitro as well as within the cellular environment. PMID:21936009

  6. PUMA promotes Bax translocation by competitive binding to Bcl-Xl during UV-induced apoptosis

    Zhang, Yingjie; Xing, Da; Wu, Yinyuan; Liu, Lei

    2008-02-01

    Ultraviolet (UV) irradiation can induce apoptosis through both the membrane death receptor and the intrinsic apoptotic signaling pathways as DNA-damaging agents. PUMA, a BH3-only Bcl-2 family protein, plays an essential role in DNA damage-induced apoptosis. Bax, also a Bcl-2 family member, translocates from the cytosol to the mitochondrial membrane during UV-induced apoptosis. However, the regulation of Bax activation induced by UV irradiation remains poorly understood. In this study, the FRET (fluorescence resonance energy transfer) technique was used to study the interactions of Bax, Bcl-Xl, and PUMA in ASTC-a-1 cells. The results show that Bax translocated from the cytosol to the mitochondrial membrane at about 7 h after UV irradiation, and the translocation can not be blocked completely when overexpressed Bcl-xl. Moreover, The interaction of Bax and Bcl-Xl weakened markedly. In addition, Co-immunoprecipitation shows that PUMA released Bax by directly binding to Bcl-XL after UV irradiation in ASTC-a-1 cells. Taken together, these results indicated that PUMA can promote Bax translocation by binding to Bcl-Xl during UV-induced apoptosis.

  7. Bax-PGAM5L-Drp1 complex is required for intrinsic apoptosis execution.

    Xu, Wenjuan; Jing, Linlin; Wang, Quanshi; Lin, Chung-Chih; Chen, Xiaoting; Diao, Jianxin; Liu, Yuanliang; Sun, Xuegang

    2015-10-01

    Intrinsic apoptosis eliminates cells with damaged DNA and cells with dysregulated expression of oncogene. PGAM5, a member of the phosphoglycerate mutase family, has two splicing variants: PGAM5L (the long form) and PGAM5S (the short form). It has been well established that PGAM5 is at the convergent point of multiple necrosis pathways. However, the role of PGAM5 in intrinsic apoptosis is still controversial. Here we report that the PGAM5L, but not PGAM5S is a prerequisite for the activation of Bax and dephosphorylation of Drp1 in arenobufagin and staurosporine induced intrinsic apoptosis. Knockdown of PGAM5L inhibits the translocation of Bax to the mitochondria and reduces mitochondrial fission. The interaction between PGAM5L and Drp1 was observed in both arenobufagin and staurosporine treated HCT116 cells, but not in HCT116 Bax(-/-) cells. Bax transfection rescues the formation of the triplex in both arenobufagin and staurosporine stimulated HCT116 Bax(-/-) cells. Arenobufagin shows remarkable anti-cancer effects both in orthotropic and heterotropic CRC models and demonstrates less toxic effects as compared with that of cisplatin. Bax-PGAM5L-Drp1 complex is detected in arenobufagin and staurosporine treated CRC cells in vitro and in arenobufagin and cisplatin treated tumor in vivo as well. In summary, our results demonstrate that Bax-PGAM5L-Drp1 complex is required for intrinsic apoptosis execution. PMID:26356820

  8. Enhancing terpenoid indole alkaloid production by inducible expression of mammalian Bax in Catharanthus roseus cells

    XU MaoJun; DONG JuFang

    2007-01-01

    Bax, a mammalian pro-apoptotic member of the Bcl-2 family, triggers hypersensitive reactions when expressed in plants. To investigate the effects of Bax on the biosynthesis of clinically important natural products in plant cells, we generate transgenic Catharanthus roseus cells overexpressing a mouse Bax protein under the β-estradiol-inducible promoter. The expression of Bax in transgenic Catharanthus roseus cells is highly dependent on β-estradiol concentrations applied. Contents of catharanthine and total terpenoid indole alkaloid of the transgenic cells treated with 30 μmol/L β-estradiol are 5.0- and 5.5-fold of the control cells. Northern and Western blotting results show that expression of mammalian Bax induces transcriptional activation of Tdc and Str, two key genes in terpenoid indole alkaloid biosynthetic pathway of Catharanthus roseus cells, and stimulates the accumulation of defense-related protein PR1 in the cells, showing that the mouse Bax triggers the defense responses of Catharanthus roseus cells and activates the terpenoid indole alkaloid biosynthetic pathway. Thus, our data suggest that the mammalian Bax might be a potential regulatory factor for secondary metabolite biosynthesis in plant cells and imply a new secondary metabolic engineering strategy for enhancing the metabolic flux to natural products by activating the whole biosynthetic pathway rather than by engineering the single structural genes within the pathways.

  9. Enhancing terpenoid indole alkaloid production by inducible expression of mammalian Bax in Catharanthus roseus cells

    2007-01-01

    Bax,a mammalian pro-apoptotic member of the Bcl-2 family,triggers hypersensitive reactions when expressed in plants.To investigate the effects of Bax on the biosynthesis of clinically important natural products in plant cells,we generate transgenic Catharanthus roseus cells overexpressing a mouse Bax protein under the β-estradiol-inducible promoter.The expression of Bax in transgenic Catharanthus roseus cells is highly dependent on β-estradiol concentrations applied.Contents of catharanthine and total terpenoid indole alkaloid of the transgenic cells treated with 30 μmol/L β-estradiol are 5.0-and 5.5-fold of the control cells.Northern and Western blotting results show that expression of mammalian Bax induces transcriptional activation of Tdc and Str,two key genes in terpenoid indole alkaloid bio-synthetic pathway of Catharanthus roseus cells,and stimulates the accumulation of defense-related protein PR1 in the cells,showing that the mouse Bax triggers the defense responses of Catharanthus roseus cells and activates the terpenoid indole alkaloid biosynthetic pathway.Thus,our data suggest that the mammalian Bax might be a potential regulatory factor for secondary metabolite biosynthesis in plant cells and imply a new secondary metabolic engineering strategy for enhancing the metabolic flux to natural products by activating the whole biosynthetic pathway rather than by engineering the single structural genes within the pathways.

  10. The effect of radiation on bcl-2 and bax in hyperplastic prostatic tissues

    Aim: To investigate the expressions of bcl-2 and bax in benign prostatic hyperplasia (BPH) and the effect of β-rays on bcl-2 and bax. Methods: The expressions of bcl-2 and bax are studied by means of immunohistochemical method in 9 normal prostate (NP) and 15 BPH and 35 patients treated with 90Sr/90Y Prostatic Hyperplasia Applicator. Results: The expressions of bcl-2 in epithelia of NP and BPH are higher than that in stroma P<0.01=. The expressions of bcl-2 in epithelia and stroma of BPH are higher than that in NP P<0.01=. The expressions of bax in epithelia of NP are higher than that in BPH P<0.05=. However ,the expressions of bcl-2 in epithelia and stroma of BPH are higher than bax P<0.01 =. Compared with the control group, the expressions of bcl-2 in epithelia and stroma of BPH treated with 90Sr/90Y Prostatic Hyperplasia Applicator decreased and the expressions of bax increased P<0.01=. Conclusion: bcl-2 gene and bax gene play an important role in the regulation of prostatic apoptosis and the treatment of β-rays can accelerate the apoptosis of prostatic tissues. (authors)

  11. A Novel Mechanism for CTCF in the Epigenetic Regulation of Bax in Breast Cancer Cells

    Claudia Fabiola Méndez-Catalá

    2013-08-01

    Full Text Available We previously reported the association of elevated levels of the multifunctional transcription factor, CCCTC binding factor (CTCF, in breast cancer cells with the specific anti-apoptotic function of CTCF. To understand the molecular mechanisms of this phenomenon, we investigated regulation of the human Bax gene by CTCF in breast and non-breast cells. Two CTCF binding sites (CTSs within the Bax promoter were identified. In all cells, breast and non-breast, active histone modifications were present at these CTSs, DNA harboring this region was unmethylated, and levels of Bax mRNA and protein were similar. Nevertheless, up-regulation of Bax mRNA and protein and apoptotic cell death were observed only in breast cancer cells depleted of CTCF. We proposed that increased CTCF binding to the Bax promoter in breast cancer cells, by comparison with non-breast cells, may be mechanistically linked to the specific apoptotic phenotype in CTCF-depleted breast cancer cells. In this study, we show that CTCF binding was enriched at the Bax CTSs in breast cancer cells and tumors; in contrast, binding of other transcription factors (SP1, WT1, EGR1, and c-Myc was generally increased in non-breast cells and normal breast tissues. Our findings suggest a novel mechanism for CTCF in the epigenetic regulation of Bax in breast cancer cells, whereby elevated levels of CTCF support preferential binding of CTCF to the Bax CTSs. In this context, CTCF functions as a transcriptional repressor counteracting influences of positive regulatory factors; depletion of breast cancer cells from CTCF therefore results in the activation of Bax and apoptosis.

  12. bax, but not bcl-2, influences the prognosis of human pancreatic cancer

    Friess, H; Lu, Z; H. Graber; Zimmermann, A.; Adler, G.; Korc, M.; Schmid, R; Buchler, M

    1998-01-01

    Background—bcl-2 and bax belong to the bcl-2-related gene family, which marks a new class of genes that influence apoptosis. The bcl-2 oncogene acts as a broad antiapoptotic factor and extends both normal and tumour cell survival. In contrast, the bax gene is a promoter of apoptosis. 
Aims—To analyse the expression of bcl-2 and bax in pancreatic cancer and correlate the results with clinical parameters. 
Patients—Pancreatic cancer tissue samples were obtained fro...

  13. Bax translocation into mitochondria during dihydroartemisinin(DHA)-induced apoptosis in human lung adenocarcinoma cells

    Lu, Ying-ying; Chen, Tong-sheng; Qu, Jun-Le

    2009-02-01

    Dihydroartemisinin (DHA), a semi-synthetic derivative of artemisinin, isolated from the traditional Chinese herb Artemisia annua, has been shown to possess promising anticancer activities and induce cancer cell death through apoptotic pathways. However, the molecular mechanisms are not well understood. This study was investigated in human lung adenocarconoma ASTC-a-1 cell line and aimed to determine whether the apoptotic process was mediated by Bax activation and translocation during DHA-induced apoptosis. In this study, DHA induced a time-dependent apoptotic cell death, which was assayed by Cell Counting Kit (CCK-8) and Hoechst 33258 staining. Detection of Bax aggregation and translocation to mitochondria was observed in living cells which were co-transfected with GFP-Bax and Dsred-mito plasmid using confocal fluorescence microscope technique. Overall, these results demonstrated that Bax activation and translocation to mitochondria occurred during DHA-induced apoptosis.

  14. Conductivity and infrared absorption of La1-xBaxCoO3 conductive ceramics

    In the present paper, the conductivity and infrared absorption of La1-xBaxCoO3 conductive ceramics have been investigated. Results show that with increasing x, the resistivity decreases, undergoes a minimum at x = 0.5, and then increases with further increases of x. The conductive ceramic with the largest infrared absorption peak corresponds to the smallest resistivity. Meanwhile the conducting mechanism of La1-xBaxCoO3 conductive ceramics have been investigated

  15. THE EXPRESSION AND CLINICAL VALUE OF APOPTOSIS CONTROL GENE Bcl-2 AND Bax IN BREAST CANCER

    ZHENG Jun; YAO Zhen-xiang; ZHANG Jing

    1999-01-01

    Objective: To study the expression and clinical value of apoptosis control gene bcl-2 and bax in breast cancer.Methods: Protein bax and bcl-2 in 41 breast cancers obtained from operations in our hospital in 1996 were detected using ABC immunohistochemical stain assay and compared with 10 cases with normal breast tissues.Results: The positive rate of bax in normal breast tissue was 90% and in breast cancer was 59%, with a significant statistical difference between them (P<0.05), but there was no statistical difference in bcl-2 protein expression. Among the 41 breast cancer, the group with lymph node metastasis (21 cases) had obviously low bax expression (43%) and high bcl-2 expression (76%), showing significant difference to the group without lymph node metastasis (P<0.05).Conclusion: The antiapoptosis function of bcl-2 was stronger than bax in breast cancer. Protein bax and bcl-2 assay may be useful in understanding the biological behaviors of breast cancer.

  16. Driving p53 Response to Bax Activation Greatly Enhances Sensitivity to Taxol by Inducing Massive Apoptosis

    Paola De Feudis

    2000-05-01

    Full Text Available The proapoptotic gene bax is one of the downstream effectors of p53. The p53 binding site in the bax promoter is less responsive to p53 than the one in the growth arrest mediating gene p21. We introduced the bax gene under the control of 13 copies of a strong p53 responsive element into two ovarian cancer cell lines. The clones expressing bax under the control of p53 obtained from the wild-type (wt p53-expressing cell line A2780 were much more sensitive (500- to 1000-fold to the anticancer agent taxol than the parent cell line, with a higher percentage of cells undergoing apoptosis after drug treatment that was clearly p53-dependent and bax-mediated. Xenografts established in nude mice from one selected clone (A2780/C3 were more responsive to taxol than the parental line and the apoptotic response of A2780/C3 tumors was also increased after treatment. Introduction of the same plasmid into the p53 null SKOV3 cell line did not alter the sensitivity to taxol or the induction of apoptosis. In conclusion, driving the p53 response (after taxol treatment by activating the bax gene rather than the p21 gene results in induction of massive apoptosis, in vitro and in vivo, and greatly enhances sensitivity to the drug.

  17. Clinical significance of bax/bcl-2 ratio in chronic lymphocytic leukemia

    Del Principe, Maria Ilaria; Bo, Michele Dal; Bittolo, Tamara; Buccisano, Francesco; Rossi, Francesca Maria; Zucchetto, Antonella; Rossi, Davide; Bomben, Riccardo; Maurillo, Luca; Cefalo, Mariagiovanna; De Santis, Giovanna; Venditti, Adriano; Gaidano, Gianluca; Amadori, Sergio; de Fabritiis, Paolo; Gattei, Valter; Del Poeta, Giovanni

    2016-01-01

    In chronic lymphocytic leukemia the balance between the pro-apoptotic and anti-apoptotic members of the bcl-2 family is involved in the pathogenesis, chemorefractoriness and clinical outcome. Moreover, the recently proposed anti-bcl-2 molecules, such as ABT-199, have emphasized the potential role of of bcl-2 family proteins in the context of target therapies. We investigated bax/bcl-2 ratio by flow cytometry in 502 patients and identified a cut off of 1.50 to correlate bax/bcl-2 ratio with well-established clinical and biological prognosticators. Bax/bcl-2 was 1.50 or over in 263 patients (52%) with chronic lymphocytic leukemia. Higher bax/bcl-2 was associated with low Rai stage, lymphocyte doubling time over 12 months, beta-2 microglobulin less than 2.2 mg/dL, soluble CD23 less than 70 U/mL and a low risk cytogenetic profile (P<0.0001). On the other hand, lower bax/bcl-2 was correlated with unmutated IGHV (P<0.0001), mutated NOTCH1 (P<0.0001) and mutated TP53 (P=0.00007). Significant shorter progression-free survival and overall survival were observed in patients with lower bax/bcl-2 (P<0.0001). Moreover, within IGHV unmutated (168 patients) and TP53 mutated (37 patients) subgroups, higher bax/bcl-2 identified cases with significant longer PFS (P=0.00002 and P=0.039). In multivariate analysis of progression-free survival and overall survival, bax/bcl-2 was an independent prognostic factor (P=0.0002 and P=0.002). In conclusion, we defined the prognostic power of bax/bcl-2 ratio, as determined by a flow cytometric approach, and highlighted a correlation with chemoresistance and outcome in chronic lymphocytic leukemia. Finally, the recently proposed new therapies employing bcl-2 inhibitors prompted the potential use of bax/bcl-2 ratio to identify patients putatively resistant to these molecules. PMID:26565002

  18. Dynamin-related protein Drp1 is required for Bax translocation to mitochondria in response to irradiation-induced apoptosis.

    Wang, Ping; Wang, Peiguo; Liu, Becky; Zhao, Jing; Pang, Qingsong; Agrawal, Samir G; Jia, Li; Liu, Feng-Ting

    2015-09-01

    Translocation of the pro-apoptotic protein Bax from the cytosol to the mitochondria is a crucial step in DNA damage-mediated apoptosis, and is also found to be involved in mitochondrial fragmentation. Irradiation-induced cytochrome c release and apoptosis was associated with Bax activation, but not mitochondrial fragmentation. Both Bax and Drp1 translocated from the cytosol to the mitochondria in response to irradiation. However, Drp1 mitochondrial translocation and oligomerization did not require Bax, and failed to induce apoptosis in Bax deficient diffuse large B-cell lymphoma (DLBCL) cells. Using fluorescent microscopy and the intensity correlation analysis, we demonstrated that Bax and Drp1 were colocalized and the levels of colocalization were increased by UV irradiation. Using co-immuno-precipitation, we confirmed that Bax and Drp1 were binding partners. Irradiation induced a time-associated increase in the interaction between active Bax and Drp1. Knocking down Drp1 using siRNA blocked UV irradiation-mediated Bax mitochondrial translocation. In conclusion, our findings demonstrate for the first time, that Drp1 is required for Bax mitochondrial translocation, but Drp1-induced mitochondrial fragmentation alone is not sufficient to induce apoptosis in DLBCL cells. PMID:26093086

  19. Dynamic interaction between 14-3-3zeta and bax during TNF-α-induced apoptosis in living cells

    Gao, Xuejuan; Xing, Da; Chen, Tongsheng

    2006-09-01

    Bax, a proapoptotic member of the Bcl-2 family, localizes largely in the cytoplasm but redistributes to mitochondria and undergoes oligomerization to induce the release of apoptogenic factors such as cytochrome c in response to apoptotic stimuli. Cytoplasmic protein 14-3-3zeta binds to Bax and, upon apoptotic stimulation, releases Bax by a caspase-independent mechanism. However, the direct interaction of the cytoplasmic 14-3-3zeta and Bax in living cells has not been observed. In present study, to monitor the dynamic interaction between 14-3-3zeta and Bax in living cells in real time during apoptosis induced by tumor necrosis factor (TNF-α), DsRed-14-3-3zeta plasmid is constructed. By cotransfecting DsRed- 14-3-3zeta and GFP-Bax plasmids into human lung adenocarcinoma cells (ASTC-a-1), we observe the dynamic interaction between Bax and 14-3-3zeta using fluorescence resonance energy transfer (FRET) technique on laser scanning confocal microscope. The results show that 14-3-3zeta remains in the cytoplasm but GFP-Bax translocates to mitochondria completely after TNF-α stimulation. These results reveal that 14-3-3zeta binds directly to Bax in healthy cells, and that 14-3-3zeta negatively regulates Bax translocation to mitochondria during TNF-α-induced apoptosis.

  20. Clinical significance of bax/bcl-2 ratio in chronic lymphocytic leukemia.

    Del Principe, Maria Ilaria; Dal Bo, Michele; Bittolo, Tamara; Buccisano, Francesco; Rossi, Francesca Maria; Zucchetto, Antonella; Rossi, Davide; Bomben, Riccardo; Maurillo, Luca; Cefalo, Mariagiovanna; De Santis, Giovanna; Venditti, Adriano; Gaidano, Gianluca; Amadori, Sergio; de Fabritiis, Paolo; Gattei, Valter; Del Poeta, Giovanni

    2016-01-01

    In chronic lymphocytic leukemia the balance between the pro-apoptotic and anti-apoptotic members of the bcl-2 family is involved in the pathogenesis, chemorefractoriness and clinical outcome. Moreover, the recently proposed anti-bcl-2 molecules, such as ABT-199, have emphasized the potential role of of bcl-2 family proteins in the context of target therapies. We investigated bax/bcl-2 ratio by flow cytometry in 502 patients and identified a cut off of 1.50 to correlate bax/bcl-2 ratio with well-established clinical and biological prognosticators. Bax/bcl-2 was 1.50 or over in 263 patients (52%) with chronic lymphocytic leukemia. Higher bax/bcl-2 was associated with low Rai stage, lymphocyte doubling time over 12 months, beta-2 microglobulin less than 2.2 mg/dL, soluble CD23 less than 70 U/mL and a low risk cytogenetic profile (Pbax/bcl-2 was correlated with unmutated IGHV (Pbax/bcl-2 (Pbax/bcl-2 identified cases with significant longer PFS (P=0.00002 and P=0.039). In multivariate analysis of progression-free survival and overall survival, bax/bcl-2 was an independent prognostic factor (P=0.0002 and P=0.002). In conclusion, we defined the prognostic power of bax/bcl-2 ratio, as determined by a flow cytometric approach, and highlighted a correlation with chemoresistance and outcome in chronic lymphocytic leukemia. Finally, the recently proposed new therapies employing bcl-2 inhibitors prompted the potential use of bax/bcl-2 ratio to identify patients putatively resistant to these molecules. PMID:26565002

  1. The associated expression of Maspin and Bax proteins as a potential prognostic factor in intrahepatic cholangiocarcinoma

    Maspin, a member of the serpin family, is a suppressor of tumor growth, an inhibitor of angiogenesis and an inducer of apoptosis. Maspin induces apoptosis by increasing Bax, a member of the Bcl-2 family of apoptosis-regulating proteins. In this exploratory study, we investigated the associated expression of Maspin and Bax proteins as a potential prognostic factor in intrahepatic cholangiocarcinoma (IHCCA). Twenty-two paraffin-embedded samples were analyzed by immunohistochemical methods using Maspin, Bax and CD34 antibodies. Maspin was scored semiquantitatively (HSCORE). Apoptosis was assessed using an antibody against cleaved caspase-3. The strong relationship observed between the expression of Maspin and Bax, indicates that Bax is likely to be the key effector of Maspin-mediated induction of apoptosis as indicated by the activation of cleaved caspase-3. We categorized Maspin HSCORE by calculating the optimal cutpoint. A Maspin HSCORE above the cutpoint was inversely related with tumor dimension, depth of tumor and vascular invasion. Uni/multivariate analysis suggests that a Maspin HSCORE below the cutpoint significantly worsens the patients' prognosis. Tumors with Maspin HSCORE below the cutpoint had a shorter survival (11+/-5 months) than did patients with Maspin HSCORE above the cutpoint (27+/-4 months), whereas Kaplan-Meier analysis and logrank test showed no significant difference in overall survival between the patients. The associated expression of Maspin and Bax might delay tumor progression in IHCCA. Maspin above the cutpoint might counteract tumor development by increasing cell apoptosis, and by decreasing tumor mass and cell invasion. The combined expression of Maspin and Bax appears to influence the susceptibility of tumor cholangiocytes to apoptosis and thus may be involved in delaying IHCCA progression

  2. The associated expression of Maspin and Bax proteins as a potential prognostic factor in intrahepatic cholangiocarcinoma

    Borghetti Angelo F

    2006-10-01

    Full Text Available Abstract Background Maspin, a member of the serpin family, is a suppressor of tumor growth, an inhibitor of angiogenesis and an inducer of apoptosis. Maspin induces apoptosis by increasing Bax, a member of the Bcl-2 family of apoptosis-regulating proteins. In this exploratory study, we investigated the associated expression of Maspin and Bax proteins as a potential prognostic factor in intrahepatic cholangiocarcinoma (IHCCA. Methods Twenty-two paraffin-embedded samples were analyzed by immunohistochemical methods using Maspin, Bax and CD34 antibodies. Maspin was scored semiquantitatively (HSCORE. Apoptosis was assessed using an antibody against cleaved caspase-3. Results The strong relationship observed between the expression of Maspin and Bax, indicates that Bax is likely to be the key effector of Maspin-mediated induction of apoptosis as indicated by the activation of cleaved caspase-3. We categorized Maspin HSCORE by calculating the optimal cutpoint. A Maspin HSCORE above the cutpoint was inversely related with tumor dimension, depth of tumor and vascular invasion. Uni/multivariate analysis suggests that a Maspin HSCORE below the cutpoint significantly worsens the patients' prognosis. Tumors with Maspin HSCORE below the cutpoint had a shorter survival (11+/-5 months than did patients with Maspin HSCORE above the cutpoint (27+/-4 months, whereas Kaplan-Meier analysis and logrank test showed no significant difference in overall survival between the patients. Conclusion The associated expression of Maspin and Bax might delay tumor progression in IHCCA. Maspin above the cutpoint might counteract tumor development by increasing cell apoptosis, and by decreasing tumor mass and cell invasion. The combined expression of Maspin and Bax appears to influence the susceptibility of tumor cholangiocytes to apoptosis and thus may be involved in delaying IHCCA progression.

  3. Bax expression measured by AQUAnalysis is an independent prognostic marker in oral squamous cell carcinoma

    Resistance to apoptosis is a hallmark of cancer and proteins regulating apoptosis have been proposed as prognostic markers in several malignancies. However, the prognostic impact of apoptotic markers has not been consistently demonstrated in oral squamous cell carcinoma (OSCC). This inconsistency in reported associations between apoptotic proteins and prognosis can be partly attributed to the intrinsic low resolution and misclassification associated with manual, semi-quantitative methods of biomarker expression measurement. The aim of this study was to examine the association between apoptosis-regulating proteins and clinical outcomes in oral squamous cell carcinoma (OSCC) using the quantitative fluorescence immunohistochemistry (IHC) based AQUAnalysis technique. Sixty-nine OSCC patients diagnosed between 1998–2005 in Calgary, Alberta, Canada were included in the study. Clinical data were obtained from the Alberta Cancer Registry and chart review. Tissue microarrays (TMAs) were assembled from triplicate cores of formalin-fixed paraffin embedded pre-treatment tumour tissue. Bax, Bcl-2 and Bcl-XL protein expression was quantified using fluorescent IHC and AQUA technology in normal oral cavity squamous epithelium (OCSE) and OSCC tumour samples. Survival was analyzed using Kaplan-Meier plots and the Cox proportional hazard model. Bax expression was predominantly nuclear in OCSE and almost exclusively cytoplasmic in OSCC. No similar differences in localization were observed for Bcl-2 or Bcl-XL. Only Bax expression associated with disease-specific survival (DSS), with 5-year survival estimates of 85.7% for high Bax versus 50.3% for low Bax (p = 0.006), in univariate analysis. High Bax expression was also significantly associated with elevated Ki67 expression, indicating that increased proliferation might lead to an improved response to radiotherapy in patients with elevated Bax expression. In multivariate analyses, Bax protein expression remained an independent

  4. Bax expression measured by AQUAnalysis is an independent prognostic marker in oral squamous cell carcinoma

    Bose Pinaki

    2012-08-01

    Full Text Available Abstract Background Resistance to apoptosis is a hallmark of cancer and proteins regulating apoptosis have been proposed as prognostic markers in several malignancies. However, the prognostic impact of apoptotic markers has not been consistently demonstrated in oral squamous cell carcinoma (OSCC. This inconsistency in reported associations between apoptotic proteins and prognosis can be partly attributed to the intrinsic low resolution and misclassification associated with manual, semi-quantitative methods of biomarker expression measurement. The aim of this study was to examine the association between apoptosis-regulating proteins and clinical outcomes in oral squamous cell carcinoma (OSCC using the quantitative fluorescence immunohistochemistry (IHC based AQUAnalysis technique. Methods Sixty-nine OSCC patients diagnosed between 1998–2005 in Calgary, Alberta, Canada were included in the study. Clinical data were obtained from the Alberta Cancer Registry and chart review. Tissue microarrays (TMAs were assembled from triplicate cores of formalin-fixed paraffin embedded pre-treatment tumour tissue. Bax, Bcl-2 and Bcl-XL protein expression was quantified using fluorescent IHC and AQUA technology in normal oral cavity squamous epithelium (OCSE and OSCC tumour samples. Survival was analyzed using Kaplan-Meier plots and the Cox proportional hazard model. Results Bax expression was predominantly nuclear in OCSE and almost exclusively cytoplasmic in OSCC. No similar differences in localization were observed for Bcl-2 or Bcl-XL. Only Bax expression associated with disease-specific survival (DSS, with 5-year survival estimates of 85.7% for high Bax versus 50.3% for low Bax (p = 0.006, in univariate analysis. High Bax expression was also significantly associated with elevated Ki67 expression, indicating that increased proliferation might lead to an improved response to radiotherapy in patients with elevated Bax expression. In multivariate analyses

  5. Identification of Bax-interacting proteins in oligodendrocyte progenitors during glutamate excitotoxicity and perinatal hypoxia–ischemia

    Sopio Simonishvili

    2013-12-01

    Full Text Available OPC (oligodendrocyte progenitor cell death contributes significantly to the pathology and functional deficits following hypoxic-ischemic injury in the immature brain and to deficits resulting from demyelinating diseases, trauma and degenerative disorders in the adult CNS. Glutamate toxicity is a major cause of oligodendroglial death in diverse CNS disorders, and previous studies have demonstrated that AMPA/kainate receptors require the pro-apoptotic protein Bax in OPCs undergoing apoptosis. The goal of the present study was to define the pro-apoptotic and anti-apoptotic effectors that regulate Bax in healthy OPCs and after exposure to excess glutamate in vitro and following H–I (hypoxia–ischemia in the immature rat brain. We show that Bax associates with a truncated form of Bid, a BH3-only domain protein, subsequent to glutamate treatment. Furthermore, glutamate exposure reduces Bax association with the anti-apoptotic Bcl family member, Bcl-xL. Cell fractionation studies demonstrated that both Bax and Bid translocate from the cytoplasm to mitochondria during the early stages of cell death consistent with a role for Bid as an activator, whereas Bcl-xL, which normally complexes with both Bax and Bid, disassociates from these complexes when OPCs are exposed to excess glutamate. Bax remained unactivated in the presence of insulin-like growth factor-1, and the Bcl-xL complexes were protected. Our data similarly demonstrate loss of Bcl-xL–Bax association in white matter following H–I and implicate active Bad in Bax-mediated OPC death. To identify other Bax-binding partners, we used proteomics and identified cofilin as a Bax-associated protein in OPCs. Cofilin and Bax associated in healthy OPCs, whereas the Bax–cofilin association was disrupted during glutamate-induced OPC apoptosis.

  6. Apoptosis repressor with a CARD domain (ARC restrains Bax-mediated pathogenesis in dystrophic skeletal muscle.

    Jennifer Davis

    Full Text Available Myofiber wasting in muscular dystrophy has largely been ascribed to necrotic cell death, despite reports identifying apoptotic markers in dystrophic muscle. Here we set out to identify the contribution of canonical apoptotic pathways to skeletal muscle degeneration in muscular dystrophy by genetically deleting a known inhibitor of apoptosis, apoptosis repressor with a card domain (Arc, in dystrophic mouse models. Nol3 (Arc protein genetic deletion in the dystrophic Sgcd or Lama2 null backgrounds showed exacerbated skeletal muscle pathology with decreased muscle performance compared with single null dystrophic littermate controls. The enhanced severity of the dystrophic phenotype associated with Nol3 deletion was caspase independent but dependent on the mitochondria permeability transition pore (MPTP, as the inhibitor Debio-025 partially rescued skeletal muscle pathology in Nol3 (-/- Sgcd (-/- double targeted mice. Mechanistically, Nol3 (-/- Sgcd (-/- mice showed elevated total and mitochondrial Bax protein levels, as well as greater mitochondrial swelling, suggesting that Arc normally restrains the cell death effects of Bax in skeletal muscle. Indeed, knockdown of Arc in mouse embryonic fibroblasts caused an increased sensitivity to cell death that was fully blocked in Bax Bak1 (genes encoding Bax and Bak double null fibroblasts. Thus Arc deficiency in dystrophic muscle exacerbates disease pathogenesis due to a Bax-mediated sensitization of mitochondria-dependent death mechanisms.

  7. Bax Inhibitor-1 down-regulation in the progression of chronic liver diseases

    Burra Patrizia

    2010-04-01

    Full Text Available Abstract Background Bax inhibitor-1 (BI-1 is an evolutionary conserved endoplasmic reticulum protein that, when overexpressed in mammalian cells, suppresses the apoptosis induced by Bax, a pro-apoptotic member of the Bcl-2 family. The aims of this study were: (1 to clarify the role of intrinsic anti- and pro-apoptotic mediators, evaluating Bax and BI-1 mRNA and protein expressions in liver tissues from patients with different degrees of liver damage; (2 to determine whether HCV and HBV infections modulate said expression. Methods We examined 62 patients: 39 with chronic hepatitis (CH (31 HCV-related and 8 HBV-related; 7 with cirrhosis (6 HCV-related and 1 HBV-related; 13 with hepatocellular carcinoma (HCC [7 in viral cirrhosis (6 HCV- and 1 HBV-related, 6 in non-viral cirrhosis]; and 3 controls. Bax and BI-1 mRNAs were quantified by real-time PCR, and BI-1 protein expression by Western blot. Results CH tissues expressed significantly higher BI-1 mRNA levels than cirrhotic tissues surrounding HCC (P Conclusions BI-1 expression is down-regulated as liver damage progresses. The high BI-1 mRNAs levels observed in early liver disease may protect virus-infected cells against apoptosis, while their progressive downregulation may facilitate hepatocellular carcinogenesis. HCV genotype seems to have a relevant role in Bax transcript expression.

  8. A novel plant glutathione S-transferase/peroxidase suppresses Bax lethality in yeast

    Kampranis, S C; Damianova, R; Atallah, M;

    2000-01-01

    for the identification of plant genes, which inhibit either directly or indirectly the lethal phenotype of Bax. Using this method a number of cDNA clones were isolated, the more potent of which encodes a protein homologous to the class theta glutathione S-transferases. This Bax-inhibiting (BI) protein...... was expressed in Escherichia coli and found to possess glutathione S-transferase (GST) and weak glutathione peroxidase (GPX) activity. Expression of Bax in yeast decreases the intracellular levels of total glutathione, causes a substantial reduction of total cellular phospholipids, diminishes the...... mitochondrial membrane potential, and alters the intracellular redox potential. Co-expression of the BI-GST/GPX protein brought the total glutathione levels back to normal and re-established the mitochondrial membrane potential but had no effect on the phospholipid alterations. Moreover, expression of BI...

  9. Bax is not involved in the resveratrol-induced apoptosis in human lung adenocarcinoma cells

    Zhang, Wei-wei; Wang, Zhi-ping; Chen, Tong-sheng

    2010-02-01

    Resveratrol (RV) is a natural plant polyphenol widely present in foods such as grapes, wine, and peanuts. Previous studies indicate that RV has an ability to inhibit various stages of carcinogenesis and eliminate preneoplastic cells in vitro and in vivo. However, little is known about the molecular mechanism of RV-induced apoptosis in human lung adenocarcinoma (ASTC-a-1) cell. In this report, we analyzed whether Bax translocation from cytoplasm to mitochondria during RV-induced apoptosis in single living cell using onfocal microscopey. Cells were transfected with GFP-Bax plasmid. Cell counting kit (CCK-8) assay was used to assess the inhibition of RV on the cells viability. Apoptotic activity of RV was detected by Hoechst 33258 and propidium iodide (PI) staining. Our results showed that RV induced a dose-dependent apoptosis in which Bax did not translocate to mitochondrias.

  10. Evidence for an Induced-Fit Process Underlying the Activation of Apoptotic BAX by an Intrinsically Disordered BimBH3 Peptide.

    Jhong, Siao-Ru; Li, Ching-Yu; Sung, Tai-Ching; Lan, Yu-Jing; Chang, Kuo-Jung; Chiang, Yun-Wei

    2016-03-17

    Apoptotic BAX protein functions as a critical gateway to mitochondria-mediated apoptosis. A diversity of stimuli has been implicated in initiating BAX activation, but the triggering mechanism remains elusive. Here we study the interaction of BAX with an intrinsically disordered BH3 motif of Bim protein (BimBH3) using ESR techniques. Upon incubation with BAX, BimBH3 binds to BAX at helices 1/6 trigger site to initiate conformational changes of BAX, which in turn promotes the formation of BAX oligomers. The study strategy is twofold: while BAX oligomerization was monitored through spectral changes of spin-labeled BAX, the binding kinetics was studied by observing time-dependent changes of spin-labeled BimBH3. Meanwhile, conformational transition between the unstructured and structured BimBH3 was measured. We show that helical propensity of the BimBH3 is increased upon binding to BAX but is then reduced after being released from the activated BAX; the release is due to the BimBH3-induced conformational change of BAX that is a prerequisite for the oligomer assembling. Intermediate states are identified, offering a key snapshot of the coupled folding and binding process. Our results provide a quantitative mechanistic description of the BAX activation and reveal new insights into the mechanism underlying the interactions between BAX and BH3-mimetic peptide. PMID:26913490

  11. Ethanol influences on Bax associations with mitochondrial membrane proteins in neonatal rat cerebellum.

    Heaton, Marieta Barrow; Siler-Marsiglio, Kendra; Paiva, Michael; Kotler, Alexandra; Rogozinski, Jonathan; Kubovec, Stacey; Coursen, Mary; Madorsky, Vladimir

    2013-02-01

    These studies investigated interactions taking place at the mitochondrial membrane in neonatal rat cerebellum following ethanol exposure and focused on interactions between proapoptotic Bax and proteins of the permeability transition pore (PTP), voltage-dependent anion channel (VDAC) and adenine nucleotide translocator (ANT) of the outer and inner mitochondrial membranes, respectively. Cultured cerebellar granule cells were used to assess the role of these interactions in ethanol neurotoxicity. Analyses were made at the age of maximal cerebellar ethanol vulnerability (P4), compared to the later age of relative resistance (P7), to determine whether differential ethanol sensitivity was mirrored by differences in these molecular interactions. We found that, following ethanol exposure, Bax proapoptotic associations with both VDAC and ANT were increased, particularly at the age of greater ethanol sensitivity, and these interactions were sustained at this age for at least 2 h postexposure. Since Bax:VDAC interactions disrupt protective VDAC interactions with mitochondrial hexokinase (HXK), we also assessed VDAC:HXK associations following ethanol treatment and found such interactions were altered by ethanol treatment, but only at 2 h postexposure and only in the P4, ethanol-sensitive cerebellum. Ethanol neurotoxicity in cultured neuronal preparations was abolished by pharmacological inhibition of both VDAC and ANT interactions with Bax but not by a Bax channel blocker. Therefore, we conclude that, at this age, within the constraints of our experimental model, a primary mode of Bax-induced initiation of the apoptosis cascade following ethanol insult involves interactions with proteins of the PTP complex and not channel formation independent of PTP constituents. PMID:22767450

  12. Influence of Apoptin on Up-regulation of the Expression of Bad and Bax

    GUO Tai; YANG Qian

    2005-01-01

    The chicken anemia virus protein, apoptin, which manifests selectivity and specificity to tumor cells, induces a p53-independent and Bcl-2-insensitive type of apoptosis in various human tumor cells. In this study, the apoptin gene was cloned from the total DNA of chicken anemia virus, and the recombinant vector was constructed. We used oligonucleotide microarray to study the changes of four genes, including Bcl-2, Bcl-xL, Bad and Bax. The post-transfection with the recombinant was also studied. The pro-apoptotic genes(Bad and Bax) and anti-apoptosis genes(Bcl-2 and Bcl-xL) were up-regulated in contrast to the controls. According to the published data, either Bcl-2 or Bcl-xL can form non-functional heterodimers by Bad and Bax binding together, resulting in blocking partly the release of cytochrome c from mitochondria. However, apoptosis could be inhibited by neither the endogenous Bcl-xL nor Bcl-2 over-expression. The experiments show that the apoptin-induced apoptotic pathway is related to the up-regulation of Bad and Bax. Bad was up-regulated by apoptin; then this up-regulated product of Bad was in favor of displacing Bax from binding to Bcl-xL or Bcl-2. Consequently, Bax exerted a pro-apoptotic dysfunction to mitochondria, thereby inducing the release of cytochrome c. Finally, apoptin induced the apoptosis of HHCC cells. These results indicate that the oligonucleotide microarray can reveal the genes related to the apoptosis induced by apoptin in HHCC cells.

  13. PUMA Promotes Bax Translocation by Both Directly Interacting with Bax and by Competitive Binding to Bcl-XL during UV-induced Apoptosis

    Zhang, Yingjie; Xing, Da; Liu, Lei

    2009-01-01

    Cell apoptosis induced by UV irradiation is a highly complex process in which different molecular signaling pathways are involved. p53 up-regulated modulator of apoptosis (PUMA) has been proposed as an important regulator in UV irradiation-induced apoptosis. However, the molecular mechanism through which PUMA regulates apoptosis, especially how PUMA activates Bcl-2-associated X protein (Bax) in response to UV irradiation is still controversial. In this study, by using real-time single-cell an...

  14. Clinical significance of bax/bcl-2 ratio in chronic lymphocytic leukemia

    Del Principe, Maria Ilaria; Bo, Michele Dal; Bittolo, Tamara; Buccisano, Francesco; Rossi, Francesca Maria; Zucchetto, Antonella; Rossi, Davide; Bomben, Riccardo; Maurillo, Luca; Cefalo, Mariagiovanna; De Santis, Giovanna; Venditti, Adriano; Gaidano, Gianluca; Amadori, Sergio; de Fabritiis, Paolo

    2016-01-01

    In chronic lymphocytic leukemia the balance between the pro-apoptotic and anti-apoptotic members of the bcl-2 family is involved in the pathogenesis, chemorefractoriness and clinical outcome. Moreover, the recently proposed anti-bcl-2 molecules, such as ABT-199, have emphasized the potential role of of bcl-2 family proteins in the context of target therapies. We investigated bax/bcl-2 ratio by flow cytometry in 502 patients and identified a cut off of 1.50 to correlate bax/bcl-2 ratio with we...

  15. NDV-induced apoptosis in absence of Bax; evidence of involvement of apoptotic proteins upstream of mitochondria

    Molouki Aidin

    2012-08-01

    Full Text Available Abstract Background Recently it was shown that following infection of HeLa cells with Newcastle disease virus (NDV, the matrix (M protein binds to Bax and subsequently the intrinsic pathway of apoptosis is activated. Moreover, there was very little alteration on mRNA and protein levels of Bax and Bcl-2 after infection with NDV. Finding In order to further investigate the role of members of the Bcl-2 family, Bax-knockout and wild-type HCT116 cells were infected with NDV strain AF2240. Although both cells underwent apoptosis through the activation of the intrinsic pathway and the release of cytochrome c from mitochondria, the percentage of dead Bax-knockout cells was significantly lower than wt cells (more than 10% at 48 h post-infection. In a parallel experiment, the effect of NDV on HT29 cells, that are originally Bcl-2-free, was studied. Apoptosis in HT29 cells was associated with Bax redistribution from cytoplasm to mitochondria, similar to that of HeLa and wt HCT116 cells. Conclusion Although the presence of Bax during NDV-induced apoptosis contributes to a faster cell death, it was concluded that other apoptotic protein(s upstream of mitochondria are also involved since cancer cells die whether in the presence or absence of Bax. Therefore, the classic Bax/Bcl-2 ratio may not be a major determinant in NDV-induced apoptosis.

  16. Prostate-specific expression of Bax delivered by an adenoviral vector induces apoptosis in LNCaP prostate cancer cells.

    Lowe, S L; Rubinchik, S; Honda, T; McDonnell, T J; Dong, J Y; Norris, J S

    2001-09-01

    In prostate carcinoma, overexpression of the anti-apoptotic gene Bcl-2 has been found to be associated with resistance to therapies including radiation and androgen ablation. Restoring the balance of Bcl-2 family members may result in the induction of apoptosis in prostate cancer cells previously resistant to treatment. To accomplish this, a strategy involving overexpression of the pro-apoptotic gene Bax was executed. The use of cytotoxic genes such as Bax require selective expression of the gene. In this study, we examined the ability of selective expression of Bax protein directed by a prostate-specific promoter to induce apoptosis in human prostate carcinoma. A second-generation adenoviral vector was constructed with the modified prostate-specific probasin promoter, ARR2PB, directing expression of an HA-tagged Bax gene and a green fluorescent protein reporter translated from an internal ribosome entry site (ARR2PB.Bax.GFP). ARR2PB promoter activity is tightly regulated and highly prostate specific and is responsive to androgens and glucocorticoids. The prostate-specific promoter-Bax-GFP transgene cassette was inserted into a cloning site near the right inverted terminal repeat of the adenoviral vector to retain specificity of the promoter. LNCaP cells infected with Ad/ARR(2)PB.Bax.GFP showed high levels of Bax expression 48 h after infection resulting in an 85% reduction in cell viability. Importantly, LNCaP cells stably transfected to overexpress Bcl-2 showed similar patterns of cell death when infected with Ad/ARR(2)PB.Bax.GFP, an 82% reduction in cell viability seen 48 h after infection. Apoptosis was confirmed by measuring caspase activation and using the TUNEL assay. Tissue specificity was evaluated using A549 cells (lung adenocarcinoma), SK-Hep-1 (liver cancer) cells, and Hela (cervical cancer) cells which did not show detectable expression of virally delivered Bax protein or any increase in cell death. Systemic administration of Ad/ARR2PB. Bax.GFP in nude

  17. The oxidized phospholipid PazePC promotes permeabilization of mitochondrial membranes by Bax

    Lidman, M.; Pokorná, Šárka; Dingeldein, A. P. G.; Sparrman, T.; Wallgren, M.; Šachl, Radek; Hof, Martin; Gröbner, G.

    2016-01-01

    Roč. 1858, č. 6 (2016), s. 1288-1297. ISSN 0005-2736 R&D Projects: GA ČR GBP208/12/G016 Institutional support: RVO:61388955 Keywords : Apoptosis * Bax-protein * Calorimetry Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 3.836, year: 2014

  18. cBid, Bax and Bcl-xL exhibit opposite membrane remodeling activities.

    Bleicken, S; Hofhaus, G; Ugarte-Uribe, B; Schröder, R; García-Sáez, A J

    2016-01-01

    The proteins of the Bcl-2 family have a crucial role in mitochondrial outer membrane permeabilization during apoptosis and in the regulation of mitochondrial dynamics. Current models consider that Bax forms toroidal pores at mitochondria that are responsible for the release of cytochrome c, whereas Bcl-xL inhibits pore formation. However, how Bcl-2 proteins regulate mitochondrial fission and fusion remains poorly understood. By using a systematic analysis at the single vesicle level, we found that cBid, Bax and Bcl-xL are able to remodel membranes in different ways. cBid and Bax induced a reduction in vesicle size likely related to membrane tethering, budding and fission, besides membrane permeabilization. Moreover, they are preferentially located at highly curved membranes. In contrast, Bcl-xL not only counterbalanced pore formation but also membrane budding and fission. Our findings support a mechanism of action by which cBid and Bax induce or stabilize highly curved membranes including non-lamellar structures. This molecular activity reduces the energy for membrane remodeling, which is a necessary step in toroidal pore formation, as well as membrane fission and fusion, and provides a common mechanism that links the two main functions of Bcl-2 proteins. PMID:26913610

  19. Opposite role of Bax and BCL-2 in the anti-tumoral responses of the immune system

    The relative role of anti apoptotic (i.e. Bcl-2) or pro-apoptotic (e.g. Bax) proteins in tumor progression is still not completely understood. The rat glioma cell line A15A5 was stably transfected with human Bcl-2 and Bax transgenes and the viability of theses cell lines was analyzed in vitro and in vivo. In vitro, the transfected cell lines (huBax A15A5 and huBcl-2 A15A5) exhibited different sensitivities toward apoptotic stimuli. huBax A15A5 cells were more sensitive and huBcl-2 A15A5 cells more resistant to apoptosis than mock-transfected A15A5 cells (pCMV A15A5). However, in vivo, in syngenic rat BDIX, these cell lines behaved differently, as no tumor growth was observed with huBax A15A5 cells while huBcl-2 A15A5 cells formed large tumors. The immune system appeared to be involved in the rejection of huBax A15A5 cells since i) huBax A15A5 cells were tumorogenic in nude mice, ii) an accumulation of CD8+ T-lymphocytes was observed at the site of injection of huBax A15A5 cells and iii) BDIX rats, which had received huBax A15A5 cells developed an immune protection against pCMV A15A5 and huBcl-2 A15A5 cells. We show that the expression of Bax and Bcl-2 controls the sensitivity of the cancer cells toward the immune system. This sensitization is most likely to be due to an increase in immune induced cell death and/or the amplification of an anti tumour immune response

  20. IMPORTANCE OF APOPTOSIS MARKERS (MDM2, BCL-2 AND Bax) IN CONVENTIONAL RENAL CELL CARCINOMA.

    Saker, Z; Tsintsadze, O; Jiqia, I; Managadze, L; Chkhotua, A

    2015-12-01

    The goal of the current study was to analyze the expression of Bcl-2, MDM2 and Bax in benign and malignant renal tissue samples and assess their possible association with different clinical parameters. Prognostic significance of the markers in recurrence-free and cancer-specific survivals has also been evaluated. Activity of MDM2, Bcl-2 and Bax was evaluated in: 24 normal human kidney tissues resected from the patients of different ages (range: 21-80 years), and in 52 conventional RCC samples. Intensity of the markers' expression was compared between the groups and correlation was analyzed with different clinical parameters. Activity of anti-apoptotic MDM2 and Bcl-2 was significantly elevated while activity of pro-apoptotic Bax was decreased in RCC as compared with normal kidney tissues. Bax expression was positively correlated with patient age. Significant association has been detected between the evaluated markers and cancer clinical parameters like: tumor stage, grade, lymph node and distant metastases. The markers' activity was associates with the tumor morphological features, in particular: presence of tumor necrosis and microvascular invasion. Disease recurrence and 5-year patient survival were associated with the markers' activity. Cox regression analyses have shown that tumor size, pathological stage and grade are the risk factors for disease recurrence and patient death. Expression of MDM2 and Bcl-2 is significantly up-regulated, while Bax is down-regulated in RCC as compared with normal kidney tissue. Intensity of the markers'activities is associated with the tumor pathological and clinical parameters (stage, grade, lymph node and distant metastases, tumor recurrence and patient survival). Further studies with more patients and longer follow-up will uncover the clinical importance of the evaluated markers in RCC. PMID:26719546

  1. The radiosensitivity of glioblastoma cell lines after hypoxia-induced Bax expression

    Full text: Radiation therapy is the most effective treatment after surgery for patients with malignant gliomas. However, the hypoxic cells exclusive to tumor tissue have proven resistant to both radiotherapy and many forms of chemotherapy. In order to specifically target these hypoxic cells, U-251 MG and U-87 MG human glioblastoma cells were stably transfected with constructs containing the suicide gene Bax under the regulation of nine copies of hypoxia-responsive elements (HREs). During hypoxia, the transcriptional complex hypoxia-inducible-factor 1 (HIF-1) binds to HRE and facilitates the transcription of downstream genes. Previously, hypoxia-induced Bax expression in transfected U-251 and U-87 clone cells has been shown to increase cell killing. The benefits of the gene therapy could be further expanded if Bax also acted to increase the sensitivity of these clone cells to radiation. To determine whether this was the case, parent and clone cells were irradiated with graded doses of X-rays under hypoxic conditions. These cells were then left hypoxic for varying durations of time, after which they were incubated for two weeks under aerated conditions to assay for clonogenic cell survival. After less than an hour under hypoxia, both U-251 and U-87 clone cells appeared significantly more sensitive to radiation than their respective parent cells. However, after longer amounts of time under anoxia, higher surviving fractions were found in each clone that were consistent with those of their respective parent cell line, showing that potentially lethal damage repair (PLDR) had occurred in the clone cells. Parent cells did not exhibit PLDR. Results are inconclusive at this point in time. Western blot analyses detailing the amount of Bax expression at each time point as well as further research exploring different durations of hypoxia will be necessary to reveal the nature of the correlation between Bax expression and radiosensitivity. Supported by NS-42927 and CA-85356

  2. Zerumbone induced apoptosis in liver cancer cells via modulation of Bax/Bcl-2 ratio

    Azimahtol Hawariah LP

    2007-04-01

    Full Text Available Abstract Background Zerumbone is a cytotoxic component isolated from Zingiber zerumbet Smith, a herbal plant which is also known as lempoyang. This new anticancer bioactive compound from Z. zerumbet was investigated for its activity and mechanism in human liver cancer cell lines. Results Zerumbone significantly showed an antiproliferative activity upon HepG2 cells with an IC50 of 3.45 ± 0.026 μg/ml. Zerumbone was also found to inhibit the proliferation of non-malignant Chang Liver and MDBK cell lines. However the IC50 obtained was higher compared to the IC50 for HepG2 cells (> 10 μg/ml. The extent of DNA fragmentation was evaluated by the Tdt-mediated dUTP nick end labelling assay which showed that, zerumbone significantly increased apoptosis in HepG2 cells in a time-course manner. In detail, the apoptotic process triggered by zerumbone involved the up-regulation of pro-apoptotic Bax protein and the suppression of anti-apoptotic Bcl-2 protein expression. The changes that occurred in the levels of this antagonistic proteins Bax/Bcl-2, was independent of p53 since zerumbone did not affect the levels of p53 although this protein exists in a functional form. Western blotting analysis for Bax protein was further confirmed qualitatively with an immunoassay that showed the distribution of Bax protein in zerumbone-treated cells. Conclusion Therefore, zerumbone was found to induce the apoptotic process in HepG2 cells through the up and down regulation of Bax/Bcl-2 protein independently of functional p53 activity.

  3. WNT signaling controls expression of pro-apoptotic BOK and BAX in intestinal cancer

    Research highlights: → Intestinal adenomas initiated by aberrant activation of the WNT pathway displayed an increased sensitivity to apoptosis. → Expression profiling of apoptosis-related genes in ApcMin/+ mice revealed the differential expression of pro-apoptotic Bok and Bax. → APC-mutant adenomatous crypts in FAP patients showed strongly increased BAX immunoreactivity. → Blocking of β-catenin/TCF-4-mediated signaling in colon cancer cells reduced the expression of BOK and BAX. -- Abstract: In a majority of cases, colorectal cancer is initiated by aberrant activation of the WNT signaling pathway. Mutation of the genes encoding the WNT signaling components adenomatous polyposis coli or β-catenin causes constitutively active β-catenin/TCF-mediated transcription, driving the transformation of intestinal crypts to cancer precursor lesions, called dysplastic aberrant crypt foci. Deregulated apoptosis is a hallmark of adenomatous colon tissue. However, the contribution of WNT signaling to this process is not fully understood. We addressed this role by analyzing the rate of epithelial apoptosis in aberrant crypts and adenomas of the ApcMin/+ mouse model. In comparison with normal crypts and adenomas, aberrant crypts displayed a dramatically increased rate of apoptotic cell death. Expression profiling of apoptosis-related genes along the crypt-villus axis and in Apc mutant adenomas revealed increased expression of two pro-apoptotic Bcl-2 family members in intestinal adenomas, Bok and Bax. Analysis of the colon of familial adenomatous polyposis (FAP) patients along the crypt-to-surface axis, and of dysplastic crypts, corroborated this expression pattern. Disruption of β-catenin/TCF-4-mediated signaling in the colorectal cancer cell line Ls174T significantly decreased BOK and BAX expression, confirming WNT-dependent regulation in intestinal epithelial cells. Our results suggest a feedback mechanism by which uncontrolled epithelial cell proliferation in the stem

  4. Bax Loss Impairs Myc-Induced Apoptosis and Circumvents the Selection of p53 Mutations during Myc-Mediated Lymphomagenesis

    Eischen, Christine M.; Roussel, Martine F.; Korsmeyer, Stanley J.; Cleveland, John L.

    2001-01-01

    The ARF and p53 tumor suppressors mediate Myc-induced apoptosis and suppress lymphoma development in Eμ-myc transgenic mice. Here we report that the proapoptotic Bcl-2 family member Bax also mediates apoptosis triggered by Myc and inhibits Myc-induced lymphomagenesis. Bax-deficient primary pre-B cells are resistant to the apoptotic effects of Myc, and Bax loss accelerates lymphoma development in Eμ-myc transgenics in a dose-dependent fashion. Eighty percent of lymphomas arising in wild-type E...

  5. Low levels of Bax inhibitor-1 gene expression increase tunicamycin-induced apoptosis in human neuroblastoma SY5Y cells

    Dan Wu; Peirong Wang; Shiyao Wang

    2012-01-01

    A human SH-SY5Y neuroblastoma cell line with a low level of Bax inhibitor-1 expression was established by lentivirus-mediated RNA interference and fluorescence-activated cell sorting. In control SH-SY5Y cells, tunicamycin treatment induced endoplasmic reticulum stress-mediated apoptosis; however, after Bax inhibitor-1 gene knockdown, cell survival rates were significantly decreased and the degree of apoptosis was significantly increased following tunicamycin treatment. In addition, chromatin condensation and apparent apoptotic phenomena, such as marginalization and cytoplasmic vesicles, were observed. Our findings indicate that Bax inhibitor-1 can delay apoptosis induced by endoplasmic reticulum stress.

  6. Conformational Heterogeneity of Bax Helix 9 Dimer for Apoptotic Pore Formation

    Liao, Chenyi; Zhang, Zhi; Kale, Justin; Andrews, David W.; Lin, Jialing; Li, Jianing

    2016-07-01

    Helix α9 of Bax protein can dimerize in the mitochondrial outer membrane (MOM) and lead to apoptotic pores. However, it remains unclear how different conformations of the dimer contribute to the pore formation on the molecular level. Thus we have investigated various conformational states of the α9 dimer in a MOM model — using computer simulations supplemented with site-specific mutagenesis and crosslinking of the α9 helices. Our data not only confirmed the critical membrane environment for the α9 stability and dimerization, but also revealed the distinct lipid-binding preference of the dimer in different conformational states. In our proposed pathway, a crucial iso-parallel dimer that mediates the conformational transition was discovered computationally and validated experimentally. The corroborating evidence from simulations and experiments suggests that, helix α9 assists Bax activation via the dimer heterogeneity and interactions with specific MOM lipids, which eventually facilitate proteolipidic pore formation in apoptosis regulation.

  7. Conformational Heterogeneity of Bax Helix 9 Dimer for Apoptotic Pore Formation.

    Liao, Chenyi; Zhang, Zhi; Kale, Justin; Andrews, David W; Lin, Jialing; Li, Jianing

    2016-01-01

    Helix α9 of Bax protein can dimerize in the mitochondrial outer membrane (MOM) and lead to apoptotic pores. However, it remains unclear how different conformations of the dimer contribute to the pore formation on the molecular level. Thus we have investigated various conformational states of the α9 dimer in a MOM model - using computer simulations supplemented with site-specific mutagenesis and crosslinking of the α9 helices. Our data not only confirmed the critical membrane environment for the α9 stability and dimerization, but also revealed the distinct lipid-binding preference of the dimer in different conformational states. In our proposed pathway, a crucial iso-parallel dimer that mediates the conformational transition was discovered computationally and validated experimentally. The corroborating evidence from simulations and experiments suggests that, helix α9 assists Bax activation via the dimer heterogeneity and interactions with specific MOM lipids, which eventually facilitate proteolipidic pore formation in apoptosis regulation. PMID:27381287

  8. Bax translocation mediated mitochondrial apoptosis and caspase dependent photosensitizing effect of Ficus religiosa on cancer cells.

    Haneef, Jazir; Parvathy, Muraleedharan; M, Parvathy; Thankayyan R, Santhosh Kumar; Sithul, Hima; Sreeharshan, Sreeja

    2012-01-01

    The main aim of the present work was to investigate the potential effect of acetone extract of Ficus religosa leaf (FAE) in multiple apoptosis signalling in human breast cancer cells. FAE treatment significantly induced dose and time dependent, irreversible inhibition of breast cancer cell growth with moderate toxicity to normal breast epithelial cells. This observation was validated using Sulforhodamine B assay. Cell cycle analysis by Flow cytometry showed cell cycle arrest in G1 phase and induction of sub-G0 peak. FAE induced chromatin condensation and displayed an increase in apoptotic population in Annexin V-FITC/PI (Fluorescein isothiocyanate/Propidium iodide) double staining. FAE stimulated the loss of mitochondrial membrane potential in multiple breast cancer cell lines when compared to normal diploid cells. To understand the role of Bax in FAE induced apoptosis, we employed a sensitive cell based platform of MCF-7 cells expressing Bax-EGFP. Bax translocation to mitochondria was accompanied by the disruption of mitochondrial membrane potential and marked elevation in LEHDase activity (Caspase 9). Consistent with this data, FAE induced Caspase activation as evidenced by ratio change in FRET Caspase sensor expressing MCF-7 cell line and cleavage of prominent Caspases and PARP. Interestingly, FAE accelerated cell death in a mitochondrial dependent manner in continuous live cell imaging mode indicating its possible photosensitizing effect. Intracellular generation of reactive oxygen species (ROS) by FAE played a critical role in mediating apoptotic cell death and photosensitizing activity. FAE induced dose and time dependent inhibition of cancer cell growth which was associated with Bax translocation and mitochondria mediated apoptosis with the activation of Caspase 9 dependent Caspase cascade. FAE also possessed strong photosensitizing effect on cancer cell line that was mediated through rapid mitochondrial transmembrane potential loss and partial Caspase

  9. Bax translocation mediated mitochondrial apoptosis and caspase dependent photosensitizing effect of Ficus religiosa on cancer cells.

    Jazir Haneef

    Full Text Available The main aim of the present work was to investigate the potential effect of acetone extract of Ficus religosa leaf (FAE in multiple apoptosis signalling in human breast cancer cells. FAE treatment significantly induced dose and time dependent, irreversible inhibition of breast cancer cell growth with moderate toxicity to normal breast epithelial cells. This observation was validated using Sulforhodamine B assay. Cell cycle analysis by Flow cytometry showed cell cycle arrest in G1 phase and induction of sub-G0 peak. FAE induced chromatin condensation and displayed an increase in apoptotic population in Annexin V-FITC/PI (Fluorescein isothiocyanate/Propidium iodide double staining. FAE stimulated the loss of mitochondrial membrane potential in multiple breast cancer cell lines when compared to normal diploid cells. To understand the role of Bax in FAE induced apoptosis, we employed a sensitive cell based platform of MCF-7 cells expressing Bax-EGFP. Bax translocation to mitochondria was accompanied by the disruption of mitochondrial membrane potential and marked elevation in LEHDase activity (Caspase 9. Consistent with this data, FAE induced Caspase activation as evidenced by ratio change in FRET Caspase sensor expressing MCF-7 cell line and cleavage of prominent Caspases and PARP. Interestingly, FAE accelerated cell death in a mitochondrial dependent manner in continuous live cell imaging mode indicating its possible photosensitizing effect. Intracellular generation of reactive oxygen species (ROS by FAE played a critical role in mediating apoptotic cell death and photosensitizing activity. FAE induced dose and time dependent inhibition of cancer cell growth which was associated with Bax translocation and mitochondria mediated apoptosis with the activation of Caspase 9 dependent Caspase cascade. FAE also possessed strong photosensitizing effect on cancer cell line that was mediated through rapid mitochondrial transmembrane potential loss and

  10. Effects of apoptosis-related proteins caspase-3, Bax and Bcl-2 on cerebral ischemia rats

    Liu, Guangyi; Tao WANG; WANG, TINGING; Song, Jinming; Zhou, Zhen

    2013-01-01

    Neuron apoptosis is known to mediate a change of ethology following cerebral ischemia-reperfusion injury in rats. Additionally, Bcl-2, Bax and caspase-3 proteins may exert a significant effect on neuron injury. The aim of this study was to investigate the role, mechanism of action and clinical significance of these proteins in neuron apoptosis and functional impairment following cerebral ischemia-reperfusion injury in rats. Sixty male healthy adult Wistar rats were randomly assigned into cont...

  11. Effect of Exercise Training on Bcl-2 and Bax Gene Expression in the Rat Heart

    Jafari; Pourrazi; Nikookheslat; Baradaran

    2015-01-01

    Background Apoptosis or programmed cell death plays an important role in the development of cardiovascular diseases, particularly heart failure. Current evidence suggests that exercise training may alter apoptosis-related signaling in sensitive somatic tissues such as the myocardium. Objectives The aim of this study was to assess the effect of exercise training on Bcl-2 and Bax genes expression as key molecules involved in intrins...

  12. The Octyl Ester of Ginsenoside Rh2 Induces Lysosomal Membrane Permeabilization via Bax Translocation

    Fang Chen

    2016-04-01

    Full Text Available Ginsenoside Rh2 is a potential pharmacologically active metabolite of ginseng. Previously, we have reported that an octyl ester derivative of ginsenoside Rh2 (Rh2-O, has been confirmed to possess higher bioavailability and anticancer effect than Rh2 in vitro. In order to better assess the possibility that Rh2-O could be used as an anticancer compound, the underlying mechanism was investigated in this study. The present results revealed that lysosomal destabilization was involved in the early stage of cell apoptosis in HepG2 cells induced by Rh2-O. Rh2-O could induce an early lysosomal membrane permeabilization with the release of lysosomal protease cathepsins to the cytosol in HepG2 cells. The Cat B inhibitor (leu and Cat D inhibitor (pepA inhibited Rh2-O-induced HepG2 apoptosis as well as tBid production and Δφm depolarization, indicating that lysosomal permeabilization occurred upstream of mitochondrial dysfunction. In addition, Rh2-O induced a significant increase in the protein levels of DRAM1 and Bax (p < 0.05 in lysosomes of HepG2 cells. Knockdown of Bax partially inhibited Rh2-O-induced Cat D release from lysosomes. Thus it was concluded that Rh2-O induced apoptosis of HepG2 cells through activation of the lysosomal-mitochondrial apoptotic pathway involving the translocation of Bax to the lysosome.

  13. The Octyl Ester of Ginsenoside Rh2 Induces Lysosomal Membrane Permeabilization via Bax Translocation.

    Chen, Fang; Zhang, Bing; Sun, Yong; Xiong, Zeng-Xing; Peng, Han; Deng, Ze-Yuan; Hu, Jiang-Ning

    2016-01-01

    Ginsenoside Rh2 is a potential pharmacologically active metabolite of ginseng. Previously, we have reported that an octyl ester derivative of ginsenoside Rh2 (Rh2-O), has been confirmed to possess higher bioavailability and anticancer effect than Rh2 in vitro. In order to better assess the possibility that Rh2-O could be used as an anticancer compound, the underlying mechanism was investigated in this study. The present results revealed that lysosomal destabilization was involved in the early stage of cell apoptosis in HepG2 cells induced by Rh2-O. Rh2-O could induce an early lysosomal membrane permeabilization with the release of lysosomal protease cathepsins to the cytosol in HepG2 cells. The Cat B inhibitor (leu) and Cat D inhibitor (pepA) inhibited Rh2-O-induced HepG2 apoptosis as well as tBid production and Δφm depolarization, indicating that lysosomal permeabilization occurred upstream of mitochondrial dysfunction. In addition, Rh2-O induced a significant increase in the protein levels of DRAM1 and Bax (p < 0.05) in lysosomes of HepG2 cells. Knockdown of Bax partially inhibited Rh2-O-induced Cat D release from lysosomes. Thus it was concluded that Rh2-O induced apoptosis of HepG2 cells through activation of the lysosomal-mitochondrial apoptotic pathway involving the translocation of Bax to the lysosome. PMID:27120618

  14. AxBAxB… pulsed atomic layer deposition: Numerical growth model and experiments

    Muneshwar, Triratna; Cadien, Ken

    2016-02-01

    Atomic layer deposition (ALD) is widely used for the fabrication of advanced semiconductor devices and related nanoscale structures. During ALD, large precursor doses (>1000 L per pulse) are often required to achieve surface saturation, of which only a small fraction is utilized in film growth while the rest is pumped from the system. Since the metal precursor constitutes a significant cost of ALD, strategies to enhance precursor utilization are essential for the scaling of ALD processes. In the precursor reaction step, precursor physisorption is restricted by steric hindrance (mA1) from ligands on the precursor molecules. On reaction, some of these ligands are removed as by-products resulting in chemisorbed species with reduced steric hindrance (mA1 → mA2, where mA2 1, x ∈ I) short-pulses rather than a single pulse. A numerical first-order surface reaction kinetics growth model is presented and applied to study the effect of AxBAxB… pulsed ALD on the growth per cycle (GPC). The model calculations predict higher GPC for AxBAxB… pulsing than with ABAB… deposition. In agreement with the model predictions, with AxBAxB… pulsed deposition, the GPC was found to increase by ˜46% for ZrN plasma enhanced ALD (PEALD), ˜49% for HfO2 PEALD, and ˜8% for thermal Al2O3 ALD with respect to conventional ABAB… pulsed growth.

  15. Structure and electrical properties of nanocrystalline La1-xBaxFeO3 for gas sensing application

    The nanocrystalline La1-xBaxFeO3 powders with x ≤ 0.3 were prepared by a sol-gel method. X-ray diffraction patterns indicate that the nanocrystalline La1-xBaxFeO3 powders are perovskite phase with orthorhombic structure. With the increasing concentration of elemental barium, the growth of the grain size is restrained. In addition, the resistance and ethanol gas-sensing properties of La1-xBaxFeO3 based sensors strongly depend on the Ba concentration. The highest response to ethanol gas (500 ppm) is 172 at 240 deg. C for the La1-xBaxFeO3 based sensor with x = 0.25.

  16. The Expression of Apoptosis-Related Genes Bcl-2 and Bax Protein and Apoptosis Positivity in Cervical Carcinoma during Irradiation

    ZHAODongli; SHIJingsen; LIMingzhong; SONGLiping; WANGShuwen

    2005-01-01

    Objective: To evaluate the apoptosis positivity, the expression of Bcl-2. bax proteins in 30 patients with squamous cell cervix carcinoma before and after radiotherapy. Methods: By using immunohistochemical and TDT-dUTP nick end labelling techniques. 30 cases of squamous cell cervical carcinoma were analyzed. Results: The apoptosis positivity before and after irradiation was 76.7%, and 100% respectively, with the difference being significant (P<0.05); The positive rates of Bcl-2 protein before and after irradiation were 73.3% and 46.7% respectively, with the difference being significant (P<0.05): The positive rates of bax protein before and after irradiation were 86% and 100 respectively, with the difference being significant (P<0.05). Conclusion: bax and Bcl-2 protein play an important role in apoptosis induced by fractionated radiation therapy. Apoptosis induced by irradiation is contributed to upregulation of bax protein or downregulation of Bcl-2 protein.

  17. A single nucleotide polymorphism in the Bax gene promoter affects transcription and influences retinal ganglion cell death

    Sheila J Semaan

    2010-03-01

    Full Text Available Pro-apoptotic Bax is essential for RGC (retinal ganglion cell death. Gene dosage experiments in mice, yielding a single wild-type Bax allele, indicated that genetic background was able to influence the cell death phenotype. DBA/2JBax+/− mice exhibited complete resistance to nerve damage after 2 weeks (similar to Bax−/− mice, but 129B6Bax+/− mice exhibited significant cell loss (similar to wild-type mice. The different cell death phenotype was associated with the level of Bax expression, where 129B6 neurons had twice the level of endogenous Bax mRNA and protein as DBA/2J neurons. Sequence analysis of the Bax promoters between these strains revealed a single nucleotide polymorphism (T129B6 to CDBA/2J at position −515. A 1.5- to 2.5-fold increase in transcriptional activity was observed from the 129B6 promoter in transient transfection assays in a variety of cell types, including RGC5 cells derived from rat RGCs. Since this polymorphism occurred in a p53 half-site, we investigated the requirement of p53 for the differential transcriptional activity. Differential transcriptional activity from either 129B6 or DBA/2J Bax promoters were unaffected in p53−/− cells, and addition of exogenous p53 had no further effect on this difference, thus a role for p53 was excluded. Competitive electrophoretic mobility-shift assays identified two DNA–protein complexes that interacted with the polymorphic region. Those forming Complex 1 bound with higher affinity to the 129B6 polymorphic site, suggesting that these proteins probably comprised a transcriptional activator complex. These studies implicated quantitative expression of the Bax gene as playing a possible role in neuronal susceptibility to damaging stimuli.

  18. Bax-induced apoptosis shortens the life span of DNA repair defect Ku70-knockout mice by inducing emphysema.

    Matsuyama, Shigemi; Palmer, James; Bates, Adam; Poventud-Fuentes, Izmarie; Wong, Kelvin; Ngo, Justine; Matsuyama, Mieko

    2016-06-01

    Cells with DNA damage undergo apoptosis or cellular senescence if the damage cannot be repaired. Recent studies highlight that cellular senescence plays a major role in aging. However, age-associated diseases, including emphysema and neurodegenerative disorders, are caused by apoptosis of lung alveolar epithelial cells and neurons, respectively. Therefore, enhanced apoptosis also promotes aging and shortens the life span depending on the cell type. Recently, we reported that ku70(-) (/) (-)bax(-) (/) (-) and ku70(-) (/) (-)bax(+/) (-) mice showed significantly extended life span in comparison with ku70(-) (/) (-)bax(+/+) mice. Ku70 is essential for non-homologous end joining pathway for DNA double strand break repair, and Bax plays an important role in apoptosis. Our study suggests that Bax-induced apoptosis has a significant impact on shortening the life span of ku70(-) (/) (-) mice, which are defective in one of DNA repair pathways. The lung alveolar space gradually enlarges during aging, both in mouse and human, and this age-dependent change results in the decrease of respiration capacity during aging that can lead to emphysema in more severe cases. We found that emphysema occurred in ku70(-) (/) (-) mice at the age of three-months old, and that Bax deficiency was able to suppress it. These results suggest that Bax-mediated apoptosis induces emphysema in ku70(-) (/) (-) mice. We also found that the number of cells, including bronchiolar epithelial cells and type 2 alveolar epithelial cells, shows a higher DNA double strand break damage response in ku70 KO mouse lung than in wild type. Recent studies suggest that non-homologous end joining activity decreases with increased age in mouse and rat model. Together, we hypothesize that the decline of Ku70-dependent DNA repair activity in lung alveolar epithelial cells is one of the causes of age-dependent decline of lung function resulting from excess Bax-mediated apoptosis of lung alveolar epithelial cells (and their

  19. Associations of MMP-2, BAX, and Bcl-2 mRNA and Protein Expressions with Development of Atrial Fibrillation.

    Diao, Shu-Ling; Xu, Hui-Pu; Zhang, Bei; Ma, Bao-Xin; Liu, Xian-Liang

    2016-01-01

    BACKGROUND To examine changes of mRNA and protein expressions of MMP-2, Bcl-2, and BAX in atrial fibrillation (AF) patients, and investigate the correlations among these 3 biomarkers. MATERIAL AND METHODS Rheumatic heart disease patients (n=158) undergoing cardiac surgical procedures for mitral valve repair or replacement were included as the AF group (n=123), containing paroxysmal AF (n=42), persistent AF (n=36), and permanent AF (n=45). Rheumatic heart disease patients with sinus rhythm (SR) (n=35) were enrolled as the SR group (control group). Immunohistochemistry, Western blot, and real-time polymerase chain reaction (PCR) were applied to detect the protein and mRNA expression levels of MMP-2, Bcl-2, and BAX. Apoptosis was observed with light and electron microscopes and detected by TdT-mediated dUTP nick-end labeling (TUNEL). RESULTS Compared with the SR group, the left atrial diameters (LADs), protein and mRNA expression levels of MMP-2 and BAX, apoptotic index (AI), and Bcl-2/BAX ratio were evidently increased in the 3 AF groups, but protein and mRNA expression levels of Bcl-2 decreased in the AF groups (all P<0.05). Correlation analysis found that MMP-2 protein expression levels was positively correlated with BAX expression, but negatively correlated with Bcl-2 expression levels. CONCLUSIONS Our study results suggest that elevated MMP-2 expression and disturbance balance of Bcl-2/BAX expressions may be associated with the development and maintenance of AF. MMP-2 may be involved in the development of AF through promoting BAX expressions and inhibiting Bcl-2. PMID:27141955

  20. Expressão da proteína Bax no tecido mamário normal de mulheres no menacme tratadas com raloxifeno Expression of Bax protein in normal tissue of premenopausal women treated with raloxifene

    Ana Maria Furtado-Veloso

    2008-04-01

    Full Text Available OBJETIVO: avaliar a expressão do antígeno Bax no epitélio mamário normal de mulheres na pré-menopausa tratadas com raloxifeno. MÉTODOS: estudo randomizado duplo-cego, envolvendo 33 mulheres pré-menopáusicas com fibroadenoma. As pacientes foram divididas em dois grupos: Placebo, (n=18 e Raloxifeno 60 mg, (n=15. A medicação foi usada durante 22 dias, começando no primeiro dia do ciclo menstrual. Uma biópsia foi realizada no 23º dia do ciclo menstrual, durante a qual uma amostra do tecido mamário normal adjacente ao fibroadenoma foi coletada e submetida a estudo imuno-histoquímico utilizando o anticorpo policlonal anti-Bax para avaliar a expressão da proteína Bax. A imunorreação para a proteína Bax foi avaliada, levando-se em consideração a intensidade e a fração de células coradas, cuja combinação resultou em um escore final de 0 a 6. Os casos com escore final >3 foram classificados como positivos para proteína Bax. O teste do c2 foi usado para análise estatística dos dados (pPURPOSE: to evaluate the expression of Bax antigen in the normal mammary epithelium of premenopausal women treated with raloxifene. METHODS: a randomized double-blind study was conducted in 33 ovulatory premenopausal women with fibroadenoma. Patients were divided into two groups: Placebo, (n=18 and Raloxifene 60 mg, (n=15. The medication was used for 22 days, beginning on the first day of the menstrual cycle. An excisional biopsy was carried out on the 23rd day of the menstrual cycle and a sample of normal breast tissue adjacent to the fibroadenoma was collected and submitted to immunohistochemical study using anti-Bax polyclonal antibody to evaluate the expression of Bax protein. Immunoreaction for Bax was evaluated taking into consideration intensity and fraction of stained cells, whose combination resulted in a final score ranging from 0 to 6. Cases with a final score >3 were classified as positive for Bax. The c2 test was used for statistical

  1. Upregulation of Bax and Bcl-2 following prenatal cocaine exposure induces apoptosis in fetal rat brain

    DaLiao Xiao, Lubo Zhang

    2008-01-01

    Full Text Available Cocaine abuse during pregnancy has been associated with numerous adverse perinatal outcomes. Aims: The present study was to determine whether prenatal cocaine exposure induced apoptosis and the possible role of Bcl-2 family genes in the programming cell death in fetal rat brain. Main methods: Pregnant rats were treated with cocaine subcutaneously (30 & 60 mg/kg/day from day 15 to 21 of gestation. Then the fetal and maternal brains were isolated. Key findings: Cocaine produced a dose-dependent decrease in fetal brain weight and brain/body weight ratio (P<0.05. Apoptotic nuclei in fetal brain were increased from 2.6 ± 0.1 (control to 8.1± 0.6 (low dose and 10.4 ± 0.2% (high dose (P<0.05. In accordance, cocaine dose dependently increased activities of caspase-3, caspase-8, and caspase-9 (% of control in the fetal brain by 177%, 155%, 174%, respectively, at 30 mg/kg/day, and by 191%, 176%, 274%, respectively, at 60 mg/kg/day. In contrast, cocaine showed no effect on caspase activities in the maternal brain. Cocaine produced a dose-dependent increase in both Bcl-2 and Bax protein expression in the fetal brain, and increased the ratio of Bax/Bcl-2 at dose of 30 mg/kg/day (P<0.05. Significance: Our study has demonstrated that prenatal cocaine exposure induces apoptosis in the fetal brain, and suggested that up-regulating Bax/Bcl-2 gene expression may be involved in cocaine-induced apoptosis. The increased apoptosis of neuronal cells in the fetal brain is likely to play a key role in cocaine-induced neuronal defects during fetal development.

  2. BAX and tumor suppressor TRP53 are important in regulating mutagenesis in spermatogenic cells in mice.

    Xu, Guogang; Vogel, Kristine S; McMahan, C Alex; Herbert, Damon C; Walter, Christi A

    2010-12-01

    During the first wave of spermatogenesis, and in response to ionizing radiation, elevated mutant frequencies are reduced to a low level by unidentified mechanisms. Apoptosis is occurring in the same time frame that the mutant frequency declines. We examined the role of apoptosis in regulating mutant frequency during spermatogenesis. Apoptosis and mutant frequencies were determined in spermatogenic cells obtained from Bax-null or Trp53-null mice. The results showed that spermatogenic lineage apoptosis was markedly decreased in Bax-null mice and was accompanied by a significantly increased spontaneous mutant frequency in seminiferous tubule cells compared to that of wild-type mice. Apoptosis profiles in the seminiferous tubules for Trp53-null were similar to control mice. Spontaneous mutant frequencies in pachytene spermatocytes and in round spermatids from Trp53-null mice were not significantly different from those of wild-type mice. However, epididymal spermatozoa from Trp53-null mice displayed a greater spontaneous mutant frequency compared to that from wild-type mice. A greater proportion of spontaneous transversions and a greater proportion of insertions/deletions 15 days after ionizing radiation were observed in Trp53-null mice compared to wild-type mice. Base excision repair activity in mixed germ cell nuclear extracts prepared from Trp53-null mice was significantly lower than that for wild-type controls. These data indicate that BAX-mediated apoptosis plays a significant role in regulating spontaneous mutagenesis in seminiferous tubule cells obtained from neonatal mice, whereas tumor suppressor TRP53 plays a significant role in regulating spontaneous mutagenesis between postmeiotic round spermatid and epididymal spermatozoon stages of spermiogenesis. PMID:20739667

  3. Enhancing survival of mouse oocytes following chemotherapy or aging by targeting Bax and Rad51.

    Loro L Kujjo

    Full Text Available BACKGROUND: Therapeutic approaches to preserve fertility in females undergoing cancer treatments are currently ineffective. This is partly due to limited knowledge of the molecular mechanisms that injured germ cells elicit to repair damage and survive or to abort repair and activate biochemical pathways leading to death. So far, we know that following spontaneously occurring or drug-induced DNA damage, the efficiency of DNA repair is a critical determinant of the cell's fate. The protein encoded by the Rad51 gene is one of several components recruited for homologous recombination-dependent DNA double-strand break repair in both somatic cells and germ cells. Recently, we showed that microinjection of recombinant Rad51 into AKR/J mouse oocytes decreased the extent of spontaneous DNA double-strand breaks, suppressed apoptosis, and restored the developmental competence in AKR/J embryos. Herein we characterized the nature of chemotherapy-induced lesions in oocytes, and the associated individual components of the DNA damage sensor and repair apparatus. For comparison, we also assessed parallel spontaneous changes in aging oocytes. METHODS: Data collected were derived from: analysis of apoptosis; immunodepletion; oocyte microinjections; immunocytochemistry; immunofluorescence; and CHIP-like assays. RESULTS: Our data show that: (i DNA damage in oocytes can be induced by both chemotherapy and spontaneously by the aging process; (ii oocytes possess the machinery and capability for repairing such DNA damage; (iii Rad51 is a critical player in the repair of both chemotherapy-induced and spontaneously-sustained DNA damage; and (iv in response to damage, oocytes exhibit an inverse functional relationship between presence of Bax and activity of Rad51. CONCLUSION/SIGNIFICANCE: Our results establish Rad51 and/or Bax as potential candidates that can be targeted for development of individualized chemotherapeutic interventions that are effective, but minimal in

  4. Expression of Ki-67, Bcl-2 and Bax in the First Trimester Abortion Materials

    Ender DÜZCAN

    2010-01-01

    Full Text Available Objective: The aim of this study was to investigate possible similar or different mechanisms in recurrent and spontaneous abortion by evaluating immunohistochemical correlation between proliferation marker Ki-67, and apoptosis markers Bcl-2 and Bax in the fetal trophoblasts and maternal deciduas from abortion material.Material and Method: Eighty samples of curettage materials from 65 abortion patients histopathologically diagnosed “decidua showing Arias-Stella reaction and chorionic villi” or only “decidua showing Arias-Stella reaction” were included in the study. Hematoxylin&Eosin stained sections from all cases were re-evaluated and further stained immunohistochemically using antibodies against Ki-67, Bcl-2 and Bax.Results: Proliferation rate evaluated by Ki-67 expression both in the cytotrophoblastic cells and decidua was found to be significantly lower in spontaneous and recurrent abortions compared to evacuation abortion. The extent of Bcl-2 expression in syncytiotrophoblastic cells covering villous stroma was also decreased in spontaneous abortion. There were no significant differences between spontaneous and recurrent abortions in terms of Bcl-2 expression in syncytiotrophoblasts and Ki-67 proliferation index in cytotrophoblastic cells or decidua. Bax staining showed minimal decidual expression in a few spontaneous and recurrent abortions.Conclusion: We concluded that proliferation rate was decreased in fetal villous cytotrophoblasts and maternal deciduas in spontaneous and recurrent abortions. We also proposed that loss of Bcl-2 expression in syncytiotrophoblasts may cause abortion in a subset of cases. However, the data from spontaneous and recurrent abortions did not not support the presence of different mechanisms in both groups.

  5. Charakterisierung des Bax Inhibitor-1 Peptides als neues tumorassoziiertes T-Zellepitop

    König, Thomas

    2010-01-01

    In der vorliegenden Arbeit wurde das neue Bax Inihibitor-1 Peptid hinsichtlich seiner Fähigkeit untersucht, in vitro eine T Zellreaktion hervorzurufen. Dies stellt die experimentelle Voraussetzung für den späteren Einsatz in Antitumor-Vakzinierungsstudien dar. Von der Arbeitsgruppe Stevanovic wurden über 70 MHC-I-gebundene Peptide von leukämischen Blasten eines AML-Patienten isoliert und analysiert. Davon wurde das HLA A24-bindende BI-1 Peptid (AYVHMVTHF) ausgewählt, da das BI-1 Protein auf G...

  6. Proteomic Profiling of Differentially Expressed Proteins from Bax inhibitor-1 Knockout and Wild Type Mice

    Li, Bo; John C Reed; Kim, Hyung-Ryong; Chae, Han-Jung

    2012-01-01

    Bax inhibitor-1 (BI-1) is an anti-apoptotic protein located in the endoplasmic reticulum (ER). The role of BI-1 has been studied in different physiopathological models including ischemia, diabetes, liver regeneration and cancer. However, fundamental knowledge about the effects of BI-1 deletion on the proteome is lacking. To further explore this protein, we compared the levels of different proteins in bi-1−/− and bi-1+/+ mouse tissues by two-dimensional electrophoresis (2-DE) and mass spectrom...

  7. Effects of Chronic Mild Stress in Female Bax Inhibitor-1-Gene Knockout Mice

    Sui, Zhi-Yan; Chae, Han-Jung; Huang, Guang-Biao; Zhao, Tong; Shrestha Muna, Sushma; Chung, Young-Chul

    2012-01-01

    Objective The anti-apoptotic protein Bax inhibitor-1 (BI-1) is a regulator of apoptosis linked to endoplasmic reticulum (ER) stress, and BI-1-/- mice exhibit increased sensitivity to tissue damage. The purpose of this study was to investigate the role of BI-1 in the pathogenesis of chronic mild stress (CMS)-induced depression-like behaviors in BI-1-/- mice. Methods We delivered CMS for 2 or 6 weeks in BI-1-knockout and wild-type mice. Control groups of BI-1-knockout and wild-type mice were le...

  8. NFAT2 mediates high glucose-induced glomerular podocyte apoptosis through increased Bax expression

    Background: Hyperglycemia promotes podocyte apoptosis and plays a key role in the pathogenesis of diabetic nephropathy. However, the mechanisms that mediate hyperglycemia-induced podocyte apoptosis is still far from being fully understood. Recent studies reported that high glucose activate nuclear factor of activated T cells (NFAT) in vascular smooth muscle or pancreatic β-cells. Here, we sought to determine if hyperglycemia activates NFAT2 in cultured podocyte and whether this leads to podocyte apoptosis. Meanwhile, we also further explore the mechanisms of NFAT2 activation and NFAT2 mediates high glucose-induced podocyte apoptosis. Methods: Immortalized mouse podocytes were cultured in media containing normal glucose (NG), or high glucose (HG) or HG plus cyclosporine A (a pharmacological inhibitor of calcinerin) or 11R-VIVIT (a special inhibitor of NFAT2). The activation of NFAT2 in podocytes was detected by western blotting and immunofluorescence assay. The role of NFAT2 in hyperglycemia-induced podocyte apoptosis was further evaluated by observing the inhibition of NFAT2 activation by 11R-VIVIT using flow cytometer. Intracellular Ca2+ was monitored in HG-treated podcocytes using Fluo-3/AM. The mRNA and protein expression of apoptosis gene Bax were measured by real time-qPCR and western blotting. Results: HG stimulation activated NFAT2 in a time- and dose-dependent manner in cultured podocytes. Pretreatment with cyclosporine A (500 nM) or 11R-VIVIT (100 nM) completely blocked NFAT2 nuclear accumulation. Meanwhile, the apoptosis effects induced by HG were also abrogated by concomitant treatment with 11R-VIVIT in cultured podocytes. We further found that HG also increased [Ca2+]i, leading to activation of calcineurin, and subsequent increased nuclear accumulation of NFAT2 and Bax expression in cultured podocytes. Conclusion: Our results identify a new finding that HG-induced podocyte apoptosis is mediated by calcineurin/NFAT2/Bax signaling pathway, which may

  9. Temporal Alterations in Cellular Bax:Bcl-2 Ratio following Traumatic Brain Injury in the Rat

    Raghupathi, Ramesh; Strauss, Kenneth I.; Zhang, Chen; Krajewski, Stanislaw; Reed, John C.; McIntosh, Tracy K.

    2003-01-01

    Cell death/survival following CNS injury may be a result of alterations in the intracellular ratio of death and survival factors. Using immunohistochemistry, Western analysis and in situ hybridization, the expression of the anti-cell death protein, Bcl-2, and the pro-cell death protein, Bax, was evaluated following lateral fluid-percussion (FP) brain injury of moderate severity (2.3–2.6 atm) in adult male Sprague-Dawley rats. By 2 h post-injury, a marked reduction of cellular Bcl-2-immunoreac...

  10. NOXA-induced alterations in the Bax/Smac axis enhance sensitivity of ovarian cancer cells to cisplatin.

    Chao Lin

    Full Text Available Ovarian cancer is the most common cause of death from gynecologic malignancy. Deregulation of p53 and/or p73-associated apoptotic pathways contribute to the platinum-based resistance in ovarian cancer. NOXA, a pro-apoptotic BH3-only protein, is identified as a transcription target of p53 and/or p73. In this study, we found that genetic variants of Bcl-2 proteins exist among cisplatin-sensitive and -resistant ovarian cancer cells, and the responses of NOXA and Bax to cisplatin are regulated mainly by p53. We further evaluated the effect of NOXA on cisplatin. NOXA induced apoptosis and sensitized A2780s and SKOV3 cells to cisplatin in vitro and in vivo. The effects were mediated by elevated Bax expression, enhanced caspase activation, release of Cyt C and Smac into the cytosol. Furthermore, gene silencing of Bax or Smac significantly attenuated NOXA and/or cisplatin-induced apoptosis in chemosensitive A2780s cells, whereas overexpression of Bax or addition of Smac-N7 peptide significantly increased NOXA and/or cisplatin-induced apoptosis in chemoresistant SKOV3 cells. To our knowledge, these data suggest a new mechanism by which NOXA chemosensitized ovarian cancer cells to cisplatin by inducing alterations in the Bax/Smac axis. Taken together, our findings show that NOXA is potentially useful as a chemosensitizer in ovarian cancer therapy.

  11. PUMA promotes Bax translocation in FOXO3a-dependent pathway during STS-induced apoptosis

    Zhang, Yingjie; Chen, Qun

    2009-08-01

    PUMA (p53 up-regulated modulator of apoptosis, also called Bbc3) was first identified as a BH3-only Bcl-2 family protein that is transcriptionally up-regulated by p53 and activated upon p53-dependent apoptotic stimuli, such as treatment with DNA-damaging drugs or UV irradiation. Recently studies have been shown that Puma is also up-regulated in response to certain p53-independent apoptotic stimuli, such as growth factor deprivation or treatment with glucocorticoids or STS (staurosporine). However, the molecular mechanisms of PUMA up-regulation and how PUMA functions in response to p53-independent apoptotic stimuli remain poorly understood. In this study, based on real-time single cell analysis, flow cytometry and western blotting technique, we investigated the function of PUMA in living human lung adenocarcinoma cells (ASTC-a-1) after STS treatment. Our results show that FOXO3a was activated by STS stimulation and then translocated from cytosol to nucleus. The expression of PUMA was up-regulated via a FOXO3a-dependent manner after STS treatment, while p53 had little function in this process. Moreover, cell apoptosis and Bax translocation induced by STS were not blocked by Pifithrin-α (p53 inhibitor), which suggested that p53 was not involved in this signaling pathway. Taken together, these results indicate that PUMA promoted Bax translocation in a FOXO3a-dependment pathway during STS-induced apoptosis, while p53 was dispensable in this process.

  12. HIV-1 Vpr-induced apoptosis is cell cycle dependent and requires Bax but not ANT.

    Joshua L Andersen

    2006-12-01

    Full Text Available The HIV-1 accessory protein viral protein R (Vpr causes G2 arrest and apoptosis in infected cells. We previously identified the DNA damage-signaling protein ATR as the cellular factor that mediates Vpr-induced G2 arrest and apoptosis. Here, we examine the mechanism of induction of apoptosis by Vpr and how it relates to induction of G2 arrest. We find that entry into G2 is a requirement for Vpr to induce apoptosis. We investigated the role of the mitochondrial permeability transition pore by knockdown of its essential component, the adenine nucleotide translocator. We found that Vpr-induced apoptosis was unaffected by knockdown of ANT. Instead, apoptosis is triggered through a different mitochondrial pore protein, Bax. In support of the idea that checkpoint activation and apoptosis induction are functionally linked, we show that Bax activation by Vpr was ablated when ATR or GADD45alpha was knocked down. Certain mutants of Vpr, such as R77Q and I74A, identified in long-term nonprogressors, have been proposed to inefficiently induce apoptosis while activating the G2 checkpoint in a normal manner. We tested the in vitro phenotypes of these mutants and found that their abilities to induce apoptosis and G2 arrest are indistinguishable from those of HIV-1NL4-3 vpr, providing additional support to the idea that G2 arrest and apoptosis induction are mechanistically linked.

  13. Arabidopsis Bax Inhibitor-1 inhibits cell death induced by pokeweed antiviral protein in Saccharomyces cerevisae

    Birsen Çakır

    2015-02-01

    Full Text Available Apoptosis is an active form of programmed cell death (PCD that plays critical roles in the development, differentiation and resistance to pathogens in multicellular organisms. Ribosome inactivating proteins (RIPs are able to induce apoptotic cell death in mammalian cells. In this study, using yeast as a model system, we showed that yeast cells expressing pokeweed antiviral protein (PAP, a single-chain ribosome-inactivating protein, exhibit apoptotic-like features, such as nuclear fragmentation and ROS production. We studied the interaction between PAP and AtBI-1 (Arabidopsis thaliana Bax Inhibitor-1, a plant anti-apoptotic protein, which inhibits Bax induced cell death. Cells expressing PAP and AtBI-1 were able to survive on galactose media compared to PAP alone, indicating a reduction in the cytotoxicity of PAP in yeast. However, PAP was able to depurinate the ribosomes and to inhibit total translation in the presence of AtBI-1. A C-terminally deleted AtBI-1 was able to reduce the cytotoxicity of PAP. Since anti-apoptotic proteins form heterodimers to inhibit the biological activity of their partners, we used a co-immunoprecipitation assay to examine the binding of AtBI-1 to PAP. Both full length and C-terminal deleted AtBI-1 were capable of binding to PAP. These findings indicate that PAP induces cell death in yeast and AtBI-1 inhibits cell death induced by PAP without affecting ribosome depurination and translation inhibition.

  14. Killing of Brain Tumor Cells by Hypoxia-Responsive Element Mediated Expression of BAX1

    Ruan, Hangjun; Wang, Jingli; Hu, Lily; Lin, Ching-Shwun; Lamborn, Kathleen R; Deen, Dennis F

    1999-01-01

    Abstract The presence of radioresistant hypoxic cells in human brain tumors limits the overall effectiveness of conventional fractionated radiation therapy. Tumor-specific therapies that target hypoxic cells are clearly needed. We have investigated the expression of suicide genes under hypoxia by a hypoxia-responsive element (HRE), which can be activated through hypoxia-inducible factor-1 (HIF-1). We transfected plasmids containing multiple copies of HRE into U-87 MG and U-251 MG-NCI human brain tumor cells and tested their ability to induce LacZ gene expression under anoxia. Gene expression under anoxia versus oxia was increased about 12-fold for U-87 MG cells and about fourfold for U-251 MG-NCI cells. At intermediate hypoxic conditions, increased LacZ gene expression in U-87 MG cells was induced by the plasmid that contained three HREs, but not by the plasmid with two HREs. Lastly, when we placed a suicide gene BAX under the control of HREs, cells transfected with the BAX plasmids were preferentially killed through apoptosis under anoxia. Our studies demonstrate that HRE-regulated gene expression is active in brain tumor cells, and that the amount of increased gene expression obtained is dependent on the cell line, the HRE copy number, and the degree of hypoxia. PMID:10933058

  15. Killing of Brain Tumor Cells by Hypoxia-Responsive Element Mediated Expression of BAX

    Hangjun Ruan

    1999-11-01

    Full Text Available The presence of radioresistant hypoxic cells in human brain tumors limits the overall effectiveness of conventional fractionated radiation therapy. Tumor-specific therapies that target hypoxic cells are clearly needed. We have investigated the expression of suicide genes under hypoxia by a hypoxia-responsive element (HRE, which can be activated through hypoxia-inducible factor-1 (HIF-1. We transfected plasmids containing multiple copies of HIRE into U-87 MG and U-251 MG-NCI human brain tumor cells and tested their ability to induce LacZ gene expression under anoxia. Gene expression under anoxia versus oxia was increased about 12-fold for U-87 MG cells and about fourfold for U-251 MG-NCI cells. At intermediate hypoxic conditions, increased LacZ gene expression in U-87 MG cells was induced by the plasmid that contained three HREs, but not by the plasmid with two HREs. Lastly, when we placed a suicide gene BAX under the control of HREs, cells transfected with the BAX plasmids were preferentially killed through apoptosis under anoxia. Our studies demonstrate that HRE-regulated gene expression is active in brain tumor cells, and that the amount of increased gene expression obtained is dependent on the cell line, the HIRE copy number, and the degree of hypoxia.

  16. The microarray expression analysis identifies BAX as a mediator of beta-carotene effects on apoptosis.

    Bodzioch, Marek; Dembinska-Kiec, Aldona; Hartwich, Jadwiga; Lapicka-Bodzioch, Katarzyna; Banas, Agnieszka; Polus, Anna; Grzybowska, Joanna; Wybranska, Iwona; Dulinska, Joanna; Gil, Dorota; Laidler, Piotr; Placha, Wojciech; Zawada, Magdalena; Balana-Nowak, Agnieszka; Sacha, Tomasz; Kiec-Wilk, Beata; Skotnicki, Aleksander; Moehle, Christoph; Langmann, Thomas; Schmitz, Gerd

    2005-01-01

    Beta-carotene is a ubiquitous compound rich in foods. However, there are conflicting reports regarding its role in carcinogenesis. We performed a microarray expression analysis in normal [human umbilical vein endothelial cells (HUVECs)] and neoplastic (melanoma A375 and myelomonocytic leukemia U937) actively proliferating cells and found evidence that beta-carotene stimulated vital cellular functions in the former and suppressed them in the latter. These differential effects correlated with the expression of the proapoptotic BCL2-associated X protein (BAX), which was downregulated in HUVECs and upregulated in the two neoplastic cell lines. The quantitative expression analysis using real-time polymerase chain reaction largely confirmed the inhibition of B-cell CLL/lymphoma 2 (BCL2) pathway-mediated apoptosis in HUVECs and its activation in melanoma and leukemic cells. The assays for apoptosis, detecting DNA breaks and caspase activation, showed consistent proapoptotic and antiapoptotic effects in U937 and HUVEC lines, respectively. However, beta-carotene-induced expression changes of BAX and other BCL2 pathway genes did not lead to the predicted induction of apoptosis in the A375 cells. PMID:15860445

  17. CO-EXPRESSIONS OF SURVIVIN GENE,BCL-2 AND BAX PROTEINS IN OVARIAN CARCINOMA

    林蓓; 张淑兰; 赵长清

    2004-01-01

    Objective To characterize the cellular properties of ovarian cancer, we examined the correlation between the expression of apoptosis-related gene survivin and those of Bcl-2 and Bar proteins. Methods Expressions of survivin mRNA, and Bcl-2 and Bax proteins in 35 cases of ovarian carcinoma, 10 cases of borderline carcinoma, 10 cases of benign tumors and 10 cases of normal tissue were evaluated by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry SABC method, respectively. Results Expression of survivin gene was detected in a significantly greater proportion in ovarian carcinoma and borderline carcinoma than those in benign tumors and normal tissues. Although there was no relationship between expression of survivin gene and FIGO stage, histologic grade, pathological type and lymphatic metastasis, expressions of Bcl-2 and Bar proteins were positively and negatively correlated with that of survivin gene, respectively. Conclusion Survivin may play an important role in pathogenesis of ovarian carcinoma, with a synergistic role of apoptosis-related gene Bcl-2protein and an antagonistic role of Bax protein in formation and progression of ovarian carcinoma.

  18. Regulation by sphingosine 1-phosphate of Bax and Bad activities during apoptosis in a MEK-dependent manner.

    Betito, Susan; Cuvillier, Olivier

    2006-02-24

    Herein we report that the prosurvival sphingolipid sphingosine 1-phosphate regulates the activities of both Bad and Bax during apoptosis of Jurkat cells. First, sphingosine 1-phosphate treatment results in Bad inactivation via the ERK/Rsk-1 pathway. Second, sphingosine 1-phosphate blocks the translocation of Bax to the mitochondria induced by Fas ligation. MEK inhibition by PD98059 or U0126 not only abrogates sphingosine 1-phosphate-induced Bad phosphorylation, but also its cytoprotective effect. Furthermore, inhibition of both mitochondrial cytochrome c efflux and Bax translocation to the mitochondria by sphingosine 1-phosphate could be overcome by PD98059 or U0126. Hence, the MEK/ERK pathway seems to be crucial for the survival effects initiated by sphingosine 1-phosphate. PMID:16414356

  19. Effects of curcumin on hippocampal Bax and Bcl-2 expression and cognitive function of a rat model of Alzheimer's disease

    Yunliang Wang; Honglei Yin; Jiyu Lou; Bing Han; Xinyue Qin; Fanchao Meng; Shuang Geng; Yajun Liu

    2011-01-01

    We tested the effects of curcumin treatment on a rat model of Alzheimer's disease induced by beta-amlyoid (Aβ1-40) expression. We investigated alterations in the expression of the apoptosis-related genes Bax and Bcl-2 in the hippocampus, as well as changes in the spatial memory and cognitive function of the rats. Reverse transcription-polymerase chain reaction and immunohistochemistry results showed that Bax expression was remarkably decreased and Bcl-2 expression was increased in the rat Alzheimer's disease model after curcumin treatment. Morris water maze results showed that the average time of escape latency was shortened in the curcumin treated model rats. Our study shows that curcumin can significantly improve spatial learning and memory functions in rats with Aβ1-40-induced Alzheimer's disease by modulating Bax and Bcl-2 expression.

  20. Effects of genistein on neuronal apoptosis, and expression of Bcl-2 and Bax proteins in the hippocampus of ovariectomized rats

    Yun Peng; Bo Jiang; Huiling Wu; Ruchun Dai; Liming Tan

    2012-01-01

    Genistein is one of several isoflavones that has a structure similar to 17β-estradiol, has a strong antioxidant effect, and a high affinity to estrogen receptors. At 15 weeks after ovariectomy, the expression of Bcl-2 in the hippocampus of rats decreased and Bax expression increased, with an obvious upregulation of apoptosis. However, intraperitoneal injection of genistein or 17β-estradiol for 15 consecutive weeks from the second day after operation upregulated Bcl-2 protein expression, downregulated Bax protein expression, and attenuated hippocampal neuron apoptosis. Our experimental findings indicate that long-term intervention with genistein can lead to a decrease in apoptosis in hippocampal neurons following ovariectomy, upregulate the expression of Bcl-2, and downregulate the expression of Bax. In addition, genistein and 17β-estradiol play equal anti-apoptotic and neuroprotective roles.

  1. The bcl-2, bax gene expression and apoptosis of continuous low-dose-rate irradiation on PC-3 transplanting tumor

    Objective: The aim of this study was to investigate bcl-2, bax expression and apoptosis of continuous low-dose-rate irradiation on prostate cancer (PC)-3 transplanting tumor. Methods: The expression of bcl-2 and bax associated with apoptosis between experiment and control groups were analyzed using immunohistochemistry at 48, 96 and 192 h after two 125I seed sources implanting model. The correlation between apoptosis and the ratio of bax/bcl-2 was analyzed using Bi-variable linear correlation. SPSS 11.0 was used to analyse the data. Results: The bcl-2 expression in experiment group began to down-regulated significantly after 125I seed irradiation for 48 h as compared with control(t=2.500, P=0.067), though it was not reached to statistical significance. At 96 and 192 h after irradiation, significantly low expression of bcl- 2 were noted (t=4.950, 3.464; P=0.008 and 0.026). In contrast, significantly over expression of bax was noted at 48, 96 and 192 h after 12si irradiation (t=3.334,4.025,5.292;P=0.029, 0.016 and 0.006). The apoptotic index (AI) for PC-3 at 48, 96 and 192 h after 125I irradiation were 22.3%, 21.7% and 30.7%, which was significantly higher than controls when at 96 and 192 h after 125I irradiation (P= 0.016 and 0.036). Moreover, positive correlation was noted between AI and bax/bcl-2 ratio (r=0.784, P= 0.012). Conclusion: Low-dose-rate irradiation could down-regulate the expression of bcl-2, up-regulate the expression of bax and induced PC-3 cells apoptosis. (authors)

  2. Recombinant factor IX (BAX326) in previously treated paediatric patients with haemophilia B: a prospective clinical trial.

    Urasinski, T; Stasyshyn, O; Andreeva, T; Rusen, L; Perina, F G; Oh, M S; Chapman, M; Pavlova, B G; Valenta-Singer, B; Abbuehl, B E

    2015-03-01

    A newly developed recombinant factor IX (BAX326(1) ) was investigated for prophylactic use in paediatric patients aged 96% of bleeds (100% of minor, 88.9% of moderate and 100% of major bleeds); the majority (88.5%) resolved after 1-2 infusions. Longer T1/2 and lower IR were observed in younger children (<6 years) compared to those aged 6 to 12 years. BAX326 administered as prophylactic treatment as well as for controlling bleeds is efficacious and safe in paediatric patients aged <12 years with haemophilia B. PMID:25495591

  3. Simultaneous Detection of Tumor Cell Apoptosis Regulators Bcl-2 and Bax through a Dual-Signal-Marked Electrochemical Immunosensor.

    Zhou, Shiwei; Wang, Yingying; Zhu, Jun-Jie

    2016-03-30

    B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Bax) are often used to monitor the apoptosis of tumor cells and evaluate cancer drug effect. In this work, a novel sandwich-type dual-signal-marked electrochemical biosensor was fabricated for simultaneous detection of Bcl-2 and Bax proteins. Reduced graphene oxide (RGO) layers were used as substrate to immobilize Bcl-2 and Bax antibodies for further capturing target antigens. CdSeTe@CdS quantum dots (QDs) and Ag nanoclusters (NCs) with antibody modification and mesoporous silica amplification were used as signal probes, which were proportional to the amount of Bcl-2 and Bax antigens. Mesoporous SiO2 can provide a larger surface area, more effectively charged by ethylene imine polymer or poly(diallyldimethylammonium chloride) to adsorb more probes. The Bcl-2 and Bax proteins were determined indirectly by the detection of oxidation peak currents of Cd and Ag using anodic stripping voltammetry, showing a good linear relationship in the protein concentration range from 1 ng/mL to 250 ng/mL. The detection limit of trace protein level was ∼0.5 fmol. The biosensor was further introduced to investigate Bcl-2 and Bax expressions from nilotinib-treated chronic myeloid leukemia K562 cells. With the increase of drug dosage and incubation time, the up-regulation for Bax and down-regulation for Bcl-2 were observed, which indicated that the apoptosis level of K562 cells could be regulated by Bcl-2 family. The ratio of Bax/Bcl-2 was further calculated for evaluation of its drug effect and apoptosis level. The limited cell amount for detection reached less than 1 × 10(3) cells, much lower than traditional methods. Furthermore, completely independent detection step and stable acid solutions containing Ag(+) and Cd(2+) for long-time storage contribute to reducing the error from the sample differences and avoiding the potential errors from the photodegradation of fluorescent probes, enzymolysis of DNA, or inactivation of

  4. Attenuation of p53 expression and Bax down-regulation during phorbol ester mediated inhibition of apoptosis

    Meßmer, Udo K; Brüne, Bernhard

    1997-01-01

    Nitric oxide (NO) caused apoptotic cell death in murine RAW 264.7 macrophages. Associated with apoptotic morphology we observed p53 up-regulation and increased Bax expression. 12-O-tetradecanoylphorbol-13-acetate (TPA), a protein kinase C (PKC) activator potently blocked NO-induced apoptosis. To gain insights into the mechanisms involved we investigated the effect of TPA on apoptotic conveying proteins such as p53 and Bax.TPA (100 nM) attentuated p53 up-regulation elicited by the NO-releasing...

  5. E2F-1 induces melanoma cell apoptosis via PUMA up-regulation and Bax translocation

    PUMA is a pro-apoptotic Bcl-2 family member that has been shown to be involved in apoptosis in many cell types. We sought to ascertain whether induction of PUMA plays a crucial role in E2F-1-induced apoptosis in melanoma cells. PUMA gene and protein expression levels were detected by real-time PCR and Western blot in SK-MEL-2 and HCT116 cell lines after Ad-E2F-1 infection. Activation of the PUMA promoter by E2F-1 overexpression was detected by dual luciferase reporter assay. E2F-1-induced Bax translocation was shown by immunocytochemistry. The induction of caspase-9 activity was measured by caspase-9 colorimetric assay kit. Up-regulation of the PUMA gene and protein by E2F-1 overexpression was detected by real-time PCR and Western blot analysis in the SK-MEL-2 melanoma cell line. In support of this finding, we found six putative E2F-1 binding sites within the PUMA promoter. Subsequent dual luciferase reporter assay showed that E2F-1 expression could increase the PUMA gene promoter activity 9.3 fold in SK-MEL-2 cells. The role of PUMA in E2F-1-induced apoptosis was further investigated in a PUMA knockout cell line. Cell viability assay showed that the HCT116 PUMA-/- cell line was more resistant to Ad-E2F-1-mediated cell death than the HCT116 PUMA+/+ cell line. Moreover, a 2.2-fold induction of the PUMA promoter was also noted in the HCT116 PUMA+/+ colon cancer cell line after Ad-E2F-1 infection. Overexpression of a truncated E2F-1 protein that lacks the transactivation domain failed to up-regulate PUMA promoter, suggesting that PUMA may be a transcriptional target of E2F-1. E2F-1-induced cancer cell apoptosis was accompanied by Bax translocation from the cytosol to mitochondria and the induction of caspase-9 activity, suggesting that E2F-1-induced apoptosis is mediated by PUMA through the cytochrome C/Apaf-1-dependent pathway. Our studies strongly demonstrated that E2F-1 induces melanoma cell apoptosis via PUMA up-regulation and Bax translocation. The signaling

  6. Laquinimod decreases Bax expression and reduces caspase-6 activation in neurons.

    Ehrnhoefer, Dagmar E; Caron, Nicholas S; Deng, Yu; Qiu, Xiaofan; Tsang, Michelle; Hayden, Michael R

    2016-09-01

    Laquinimod is an immunomodulatory compound that has shown neuroprotective benefits in clinical trials for multiple sclerosis. Laquinimod ameliorates both white and gray matter damage in human patients, and prevents axonal degeneration in animal models of multiple sclerosis. Axonal damage and white matter loss are a common feature shared between different neurodegenerative diseases. Caspase-6 activation plays an important role in axonal degeneration on the molecular level. Increased activity of caspase-6 has been demonstrated in brain tissue from presymptomatic Huntington disease mutation carriers, and it is an early marker of axonal dysfunction. Since laquinimod is currently undergoing a clinical trial in Huntington disease (LEGATO-HD, clinicaltrials.gov ID: NCT02215616), we set out to evaluate its impact on neuronal caspase-6 activation. We find that laquinimod ameliorates DNA-damage induced activation of caspase-6 in primary neuronal cultures. This is an indirect effect that is not mediated by direct inhibition of the enzyme. The investigation of potential caspase-6 activating mechanisms revealed that laquinimod reduces the expression of Bax, a pro-apoptotic molecule that causes mitochondrial cytochrome c release and caspase activation. Bax expression is furthermore increased in striatal tissues from the YAC128 mouse model of HD in an age-dependent manner. Our results demonstrate that laquinimod can directly downregulate neuronal apoptosis pathways relevant for axonal degeneration in addition to its known effects on astrocytes and microglia in the CNS. It targets a pathway that is relevant for the pathogenesis of HD, supporting the hypothesis that laquinimod may provide clinical benefit. PMID:27296315

  7. Effect of Exercise Training on Bcl-2 and Bax Gene Expression in the Rat Heart

    Jafari

    2015-10-01

    Full Text Available Background Apoptosis or programmed cell death plays an important role in the development of cardiovascular diseases, particularly heart failure. Current evidence suggests that exercise training may alter apoptosis-related signaling in sensitive somatic tissues such as the myocardium. Objectives The aim of this study was to assess the effect of exercise training on Bcl-2 and Bax genes expression as key molecules involved in intrinsic pathway of apoptosis in the rat heart. Materials and Methods This study was conducted with a two-group experimental design (animal model and sixteen three-month-old male rats were selected and randomly divided to two groups of exercise training (n = 8 and control (n = 8. Rats in the trained group participated in an exercise training program for 12 weeks (10 – 60 m min-1, 24 – 33 min d-1, 15%. The rat hearts were removed forty-eight hours after the last training session. RNA extraction and synthesis of cDNA was done, and Bax and Bcl2 genes expression was analyzed through the Real Time-Polymerase Chain Reaction (RT-PCR. Kolmogorov-Smirnov and independent t-test were applied for statistical analysis of the data (P 0.05. However, Bcl2 expression was higher in the trained group compared to the control group (11%. Conclusions In general, it seems that three-month exercise training was effective in reducing cardiac mitochondrial pro-apoptotic protein. However, considering the results of the Bcl2 gene expression, more researches are needed to identify effects of exercise trainings on indices of myocardial apoptosis.

  8. The effect of pentoxifylline drug on bax/bcl2 gene dosage expression changes following ischemic reperfusion injury in kidney

    Mehrdad Hashemi

    2015-11-01

    Full Text Available Global cerebral ischemia (GCI and reperfusion induced apoptosis that lead to cell injury and death. The bax and bcl-2 are pro-apoptotic and anti-apoptotic genes, respectively. These genes belong to The B-cell lymphoma-2 (bcl-2 family.In this study; we assessed the effect of pentoxifylline drug on bax/bcl2 gene dosage expression changes following ischemic reperfusion injury in kidney. In this experimental study, 20 male wistar rats were accidently divided them on two tenth group of control and treatment groups. In the control group, celiotomy was performed by ventral midline incision. The left kidney was isolated, and then both the renal artery and vein were obstructed. After 60 minutes of warm ischemia, vessel obstruction resolved and the right kidney was removed. 72 hours after reperfusion, tissue samples were taken from left kidney for histopathology. All these steps in treatment group were exactly repeated after administration of 45 mg/kg/PO pentoxifylline (3 hours before operation and in this group treatment was continued every 12h until 3 days. In this research quantitative real-time PCR is used for the detection expression Bcl2 and Bax genes in ischemia group and PNT drug group and compared to normal sample. The results showed the gene dosage ratio of bax/bcl2 in PNT group decline than to ischemia group. Therefore, the pentoxyfylline might have a role in control of apoptosis result from Ischemia- reperfusion

  9. Expression of Bax in yeast affects not only the mitochondria but also vacuolar integrity and intracellular protein traffic

    Dimitrova, Irina; Toby, Garabet G; Tili, Esmerina;

    2004-01-01

    -transferase (BI-GST) leads to aggregation, but not fusion of the mitochondria. In addition, Bax affects the integrity of yeast vacuoles, resulting in the disintegration and eventual loss of the organelles, and the disruption of intracellular protein traffic. While Bcl-2 coexpression only partially corrects...

  10. Apoptosis and Bax expression are increased by coal dust in the polycyclic aromatic hydrocarbon-exposed lung

    Ghanem, M.M.; Battelli, L.A.; Mercer, R.R.; Scabilloni, J.F.; Kashon, M.L.; Ma, J.Y.C.; Nath, J.; Hubbs, A.F.

    2006-09-15

    Miners inhaling respirable coal dust (CD) frequently develop coal workers' pneumoconiosis. Many coal miners are also exposed to polycyclic aromatic hydrocarbon (PAH) components of diesel engine exhaust and cigarette smoke, which may contribute to lung disease in these workers. Recently, apoptosis was reported to play a critical role in the development of another pneumoconiosis of miners, silicosis. In addition, CID was reported to suppress cytochrome P450 1A1 (CYP1A1) induction by PAHs. We exposed rats intratracheally to 0.0, 2.5, 10.0, 20.0, or 40.0 mg/rat CD and, 11 days later, to intraperitoneal P-naphthoflavone (BNF), a PAH. In another group of rats exposed to CD and BNF, caspase activity was inhibited by injection of the pan-caspase inhibitor Q-VD-OPH (quinoline-Val-Asp (OMe)-CH{sub 2}-OPH). In rats exposed to BNF, CD exposure increased alveolar expression of the proapoptotic mediator Bax but decreased CYP1A1 induction relative to BNF exposure alone. Pan-caspase inhibition decreased CD-associated Bax expression and apoptosis but did not restore CYP1A1 activity. Further, CD-induced lung inflammation and alveolar epithelial cell hypertrophy and hyperplasia were not suppressed by caspase inhibition. It is concluded that combined BNF and CD exposure increased Bax expression and apoptosis in the lung, but Bax and apoptosis were not the major determinants of early lung injury in this model.

  11. Effect of Bcl-2/Bax gene expression on apoptosis of spermatogenic cells of mouse testes induced by low dose radiation

    The different kinds of spermatogenic cells were separated using density gradient centrifugation and their apoptosis and Bcl-2 and Bax protein expression were measured with flow cytometry and immunohistochemical method, respectively. The results showed the apoptosis in all kinds of spermatogenic cells induced by low dose radiation (LDR) had a obvious regularity. When the doses were 0.025 and 0.05 Gy, spermatogonia apoptosis was dominant. With the increase of irradiation dose (0.075-0.2 Gy), spermatocytes also showed an apoptotic change, but the apoptotic percentage of spermatogonia was significantly higher than that of spermatocytes. Moreover, the apoptosis of spermatids and spermatozoa scarcely occurred after LDR. Bax protein was primarily expressed in spermatogonia and spermatocytes, and the former was significantly higher than that of the latter after LDR. With the increase of irradiation dose, Bax protein expression showed a upgrading tendency, but that of spermatids and spermatozoa scarcely occurred. Bcl-2 protein was primarily expressed in spermatids and spermatozoa, but the Bcl-2 protein expressions of spermatogonia and spermatocytes scarcely occurred after LDR. These results imply that the interacting regulation of Bcl-2 and Bax gene expression might be involved in selective apoptosis of spermatogenic cells induced by LDR, which provided an experimental evidence for further exploring the apoptotic mechanism of adaptive response of spermatogenic cells by LDR

  12. Immunohistochemical study of epidermal and dermal expression of Bcl-X, Bcl-2 and bax in psoriasis.

    2000-01-01

    Objective: To investigate the regulation of cell proliferation and apoptosis in psoriasis. Methods: The expressions of Bcl-X, Bcl-2 and Bax were studied with immunohistochemical technique (SP) in the lesional and non-lesional psoriatic skin. Results: There were significant overexpressions of Bcl-X in all layers of epidermis, inflammatory cells and vascular endothelia in dermis;

  13. Erythropoietin can promote survival of cerebral cells by downregulating Bax gene after traumatic brain injury in rats

    Liao Z

    2009-01-01

    Full Text Available Background : Traumatic brain injury (TBI is an important cause of adult mortality and morbidity. Erythropoietin (Epo has been shown to promote the viability of cerebral cells by upregulating Bcl-2 gene; however, Epo may exert its antiapoptotic effect via the differential regulation of the expression of genes involved in the apoptotic process. Aim : The present study examined the neuroprotective effect of Epo as a survival factor through the regulation of the Bax. Materials and Methods : Wistar rats were randomly divided into three groups: Recombinant human EPO treated (rhEPO TBI, vehicle-treated TBI, and sham-operated. Traumatic brain injury was induced by the Feeney free-falling model. Rats were killed 5, 12, 24, 72, 120, or 168 h after TBI. Regulation of Bcl-2 was detected by reverse transcription-polymerase chain reaction (RT-PCR, western blotting and immunofluorescence. Results : Bax mRNA and protein levels were lower in the rhEPO-treated rat brains than in the vehicle-treated rat brains. Induction of Bax expression peaked at 24 h and remained stable for 72-120 h in vehicle-treated rat brains, whereas induction of Bax expression was only slightly elevated in rhEPO-treated rat brains. The number of TdT-mediated dUTP Nick-End Labeling(TUNEL-positive cells in the rhEPO-treated rat brains was far fewer than in the vehicle-treated rat brains. Conclusions : Epo exerts neuroprotective effect against traumatic brain injury via reducing Bax gene expression involved in inhibiting TBI-induced neuronal cell death.

  14. Bax to Bcl-2 ratio and Ki-67 index are useful predictors of neoadjuvant chemoradiation therapy in bladder cancer

    In this study, locally advanced bladder cancer was treated by radiation combined with cisplatin therapy and a retrospective analysis was conducted to predict the clinical response to chemoradiotherapy (CRT) based on the immunohistochemistry of apoptosis-related proteins. Sixty-two patients (median age, 68 years; range, 45-89 years) with transitional cell carcinoma of the bladder (pT1G3-pT4M0) treated with CRT (median dose: 40.5 Gy of radiation and 230 mg of cisplatin) were studied. Mucosal biopsy was performed before and after CRT. Paraffin-embedded tumor specimens were examined with TdT-mediated dUTP-biotin nick end-labeling (TUNEL) and were immunostained for Ki-67, p53, Bcl-2 and Bax; the Bax/Bcl-2 ratio and apoptosis index (AI) were calculated. Clinical features of the patients and response to CRT were compared with data obtained from examination of the tumors. The 62 patients had a median follow-up period of 34 months (range, 3-84 months). Responses to CRT were as follows: complete response (CR), 34%; partial response (PR), 45%; no change (NC), 21%. The survival rate of patients with Ki-67-positive tumors was significantly lower than those of patients with Ki-67-negative tumors (P<0.05). No significant correlation was observed between the expression of any protein, the AI and the clinical response. However, the Bax/Bcl-2 ratio showed a significant association with the CR rate (P=0.0289). The results of this study suggest that the combined assessment of Bcl-2 and Bax protein expression may be used to predict a clinical response to CRT based on the Bax/Bcl-2 ratio determined before therapy. The Ki-67 index may be a useful predictor of prognosis in patients treated by CRT. (author)

  15. Modification of the BAX Salmonella test kit to include a hot start functionality (modification of AOAC Official Method 2003.09).

    Wallace, F Morgan; DiCosimo, Deana; Farnum, Andrew; Tice, George; Andaloro, Bridget; Davis, Eugene; Burns, Frank R

    2011-01-01

    In 2010, the BAX System PCR assay for Salmonella was modified to include a hot start functionality designed to keep the reaction enzyme inactive until PCR begins. To validate the assay's Official Methods of Analysis status to include this procedure modification, an evaluation was conducted on four food types that were simultaneously analyzed with the BAX System and either the U.S. Food and Drug Administration's Bacteriological Analytical Manual or the U.S. Department of Agriculture-Food Safety and Inspection Service Microbiology Laboratory Guidebook reference method for detecting Salmonella. Identical performance between the BAX System method and the reference methods was observed. Additionally, lysates were analyzed using both the BAX System Classic and BAX System Q7 instruments with identical results using both platforms for all samples tested. Of the 100 samples analyzed, 34 samples were positive for both the BAX System and reference methods, and 66 samples were negative by both the BAX System and reference methods, demonstrating 100% correlation. No instrument platform variation was observed. Additional inclusivity and exclusivity testing using the modified test kit demonstrated the test kit to be 100% accurate in evaluation of test panels of 352 Salmonella strains and 46 non-Salmonella strains. PMID:22165013

  16. Effect of Achyranthes bidentata polysaccharides on the expression of BCL-2 and bax in hepatic tissues after exhaustive exercise in rats.

    Lin, Jinyang; Zhang, Zhuoying; Shan, Ying

    2010-01-01

    This study aims to assess the effects of Achyranthes bidentata polysaccharides (ABPS) on the expression of bcl-2 and bax in hepatic tissues after exhaustive exercise in order to provide theoretical support for the application of ABPS in the field of sports nutrition. Thirty male Sprague-Dawley rats were randomized into three groups, each consisting of 10 rats: Normal control group (NCG), Exhausting exercises control group (EECG), ABPS treated group (ATG). ABPS were fed orally by gastric intubation to rats of ABPS treated group (ATG) once daily for 7 days. Control animals (EECG and NCG) received the same amount of isotonic sodium chloride solution. Exhaustive exercise was performed on a rodent treadmill. The SP (streptavidin peroxidase) method for immunohistochemical staining was adopted to test the protein expression of bax and bcl-2 in the hepatic tissues of the rats. Exhausting exercises increased bax protein expression of hepatic tissues of rats and bax/bcl-2 ratio dramatically, but a decreased bcl-2 protein expression. In the rats fed ABPS orally by gastric intubation, the bax protein expression and bax/bcl-2 ratio obviously decreased, while bcl-2 protein expression increased. The result indicated that bax and bcl-2 co-regulated the exercise-induced hepatocyte apoptosis. Feeding ABPS orally by gastric intubation to rats can inhibit the hepatocyte apoptosis in exhaustive exercise. PMID:21731162

  17. Boron neutron capture therapy induces apoptosis of glioma cells through Bcl-2/Bax

    Mao Xinggang

    2010-12-01

    Full Text Available Abstract Background Boron neutron capture therapy (BNCT is an alternative treatment modality for patients with glioma. The aim of this study was to determine whether induction of apoptosis contributes to the main therapeutic efficacy of BNCT and to compare the relative biological effect (RBE of BNCT, γ-ray and reactor neutron irradiation. Methods The neutron beam was obtained from the Xi'an Pulsed Reactor (XAPR and γ-rays were obtained from [60Co] γ source of the Fourth Military Medical University (FMMU in China. Human glioma cells (the U87, U251, and SHG44 cell lines were irradiated by neutron beams at the XAPR or [60Co] γ-rays at the FMMU with different protocols: Group A included control nonirradiated cells; Group B included cells treated with 4 Gy of [60Co] γ-rays; Group C included cells treated with 8 Gy of [60Co] γ-rays; Group D included cells treated with 4 Gy BPA (p-borono-phenylalanine-BNCT; Group E included cells treated with 8 Gy BPA-BNCT; Group F included cells irradiated in the reactor for the same treatment period as used for Group D; Group G included cells irradiated in the reactor for the same treatment period as used for Group E; Group H included cells irradiated with 4 Gy in the reactor; and Group I included cells irradiated with 8 Gy in the reactor. Cell survival was determined using the 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium (MTT cytotoxicity assay. The morphology of cells was detected by Hoechst33342 staining and transmission electron microscope (TEM. The apoptosis rate was detected by flow cytometer (FCM. The level of Bcl-2 and Bax protein was measured by western blot analysis. Results Proliferation of U87, U251, and SHG44 cells was much more strongly inhibited by BPA-BNCT than by irradiation with [60Co] γ-rays (P 60Co] γ-rays (P P Conclusions Compared with ��-ray and reactor neutron irradiation, a higher RBE can be achieved upon treatment of glioma cells with BNCT. Glioma cell apoptosis induced by

  18. Titanium dioxide induces apoptotic cell death through reactive oxygen species-mediated Fas upregulation and Bax activation

    Yoon TH

    2012-03-01

    Full Text Available Ki-Chun Yoo1, Chang-Hwan Yoon1, Dongwook Kwon2, Kyung-Hwan Hyun1, Soo Jung Woo1, Rae-Kwon Kim1, Eun-Jung Lim1, Yongjoon Suh1, Min-Jung Kim1, Tae Hyun Yoon2, Su-Jae Lee11Laboratory of Molecular Biochemistry, 2Laboratory of Nanoscale Characterization and Environmental Chemistry, Department of Chemistry, Hanyang University, Seoul, Republic of KoreaBackground: Titanium dioxide (TiO2 has been widely used in many areas, including biomedicine, cosmetics, and environmental engineering. Recently, it has become evident that some TiO2 particles have a considerable cytotoxic effect in normal human cells. However, the molecular basis for the cytotoxicity of TiO2 has yet to be defined.Methods and results: In this study, we demonstrated that combined treatment with TiO2 nanoparticles sized less than 100 nm and ultraviolet A irradiation induces apoptotic cell death through reactive oxygen species-dependent upregulation of Fas and conformational activation of Bax in normal human cells. Treatment with P25 TiO2 nanoparticles with a hydrodynamic size distribution centered around 70 nm (TiO2P25–70 together with ultraviolet A irradiation-induced caspase-dependent apoptotic cell death, accompanied by transcriptional upregulation of the death receptor, Fas, and conformational activation of Bax. In line with these results, knockdown of either Fas or Bax with specific siRNA significantly inhibited TiO2-induced apoptotic cell death. Moreover, inhibition of reactive oxygen species with an antioxidant, N-acetyl-L-cysteine, clearly suppressed upregulation of Fas, conformational activation of Bax, and subsequent apoptotic cell death in response to combination treatment using TiO2P25–70 and ultraviolet A irradiation.Conclusion: These results indicate that sub-100 nm sized TiO2 treatment under ultraviolet A irradiation induces apoptotic cell death through reactive oxygen species-mediated upregulation of the death receptor, Fas, and activation of the preapoptotic protein

  19. Bcl-2 and bax expression and prostate cancer outcome in men treated with radiotherapy in Radiation Therapy Oncology Group protocol 86-10

    Purpose: Bcl-2 and bax are proteins with opposing roles in apoptosis regulation; yet abnormal expression of either has been associated with failure after radiotherapy (RT). In this study we examined bcl-2 and bax expression as predictive markers in men treated with radiotherapy ± androgen deprivation on Radiation Therapy Oncology Group (RTOG) protocol 86-10. Experimental Design: Suitable archival diagnostic tissue was obtained from 119 (26%) patients for bcl-2 analysis and 104 (23%) patients for bax analysis. Cox proportional hazards multivariate analysis was used to determine the relationship of abnormal bcl-2 and bax expression to the end points of local failure, distant metastasis, cause-specific mortality, and overall mortality. Bcl-2 overexpression was classified as any tumor cell cytoplasmic staining and altered bax expression was classified as greater or lesser cytoplasmic staining intensity of tumor cells as compared with adjacent normal prostate epithelium. Results: The study cohort exhibited bcl-2 overexpression in 26% (n = 30) of cases and abnormal bax expression in 47% (n = 49) of cases. A borderline significant relationship was observed between abnormal bax expression and higher Gleason score (p = 0.08). In univariate and multivariate analyses, there was no statistically significant relationship seen between abnormal bcl-2 or bax expression and outcome. Conclusions: Abnormal bcl-2 and bax expression were not related to any of the end points tested. The cohort examined was comprised of patients with locally advanced disease and it is possible that these markers may be of greater value in men with earlier-stage prostate cancer

  20. Characterization of BAX inhibitor-1 as a novel leukemia-associated antigen.

    Schmidt, S M; König, T; Bringmann, A; Held, S; von Schwarzenberg, K; Heine, A; Holderried, T A W; Stevanovic, S; Grünebach, F; Brossart, P

    2009-10-01

    Using dendritic cells (DCs) electroporated with whole RNA isolated from blasts of a patient with acute myeloid leukemia (AML), we were able to generate leukemia-specific cytotoxic T lymphocytes (CTLs) capable of recognizing the leucemic cells. To identify T-cell epitopes mediating lysis of malignant cells, peptides were eluted from the patient's blasts and analyzed by mass spectrometry (LC/MS)-based peptide sequencing. Using this approach, an HLA-A24-binding peptide derived from Bax inhibitor-1 (BI-1), a regulator of apoptosis pathways, was identified as an epitope recognized by the generated CTLs. To further characterize this novel antigenic peptide, CTLs were induced using DCs electroporated with RNA coding for BI-1 or pulsed with the cognate peptide. These CTLs generated from healthy donors in vitro efficiently lysed the patient's blasts as well as other HLA-matched leukemic cells. In conclusion, we identified a BI-1 peptide as a novel immunogenic tumor-associated antigen (TAA) in AML. In vitro induction of BI-1-specific CTLs by RNA transfection or pulsing of DCs with the synthetically generated peptide was a feasible and highly effective method to generate leukemia-specific CTLs. As BI-1 is (over-) expressed in a broad variety of malignancies, it may represent an interesting novel TAA in the context of cancer vaccines. PMID:19609282

  1. Expression of Arabidopsis Bax Inhibitor-1 in transgenic sugarcane confers drought tolerance.

    Ramiro, Daniel Alves; Melotto-Passarin, Danila Montewka; Barbosa, Mariana de Almeida; Santos, Flavio Dos; Gomez, Sergio Gregorio Perez; Massola Júnior, Nelson Sidnei; Lam, Eric; Carrer, Helaine

    2016-09-01

    The sustainability of global crop production is critically dependent on improving tolerance of crop plants to various types of environmental stress. Thus, identification of genes that confer stress tolerance in crops has become a top priority especially in view of expected changes in global climatic patterns. Drought stress is one of the abiotic stresses that can result in dramatic loss of crop productivity. In this work, we show that transgenic expression of a highly conserved cell death suppressor, Bax Inhibitor-1 from Arabidopsis thaliana (AtBI-1), can confer increased tolerance of sugarcane plants to long-term (>20 days) water stress conditions. This robust trait is correlated with an increased tolerance of the transgenic sugarcane plants, especially in the roots, to induction of endoplasmic reticulum (ER) stress by the protein glycosylation inhibitor tunicamycin. Our findings suggest that suppression of ER stress in C4 grasses, which include important crops such as sorghum and maize, can be an effective means of conferring improved tolerance to long-term water deficit. This result could potentially lead to improved resilience and yield of major crops in the world. PMID:26872943

  2. Direct proof of static charge stripe correlations in La2-xBaxCuO4

    Chen, X. M.; Thampy, V.; Mazzoli, C.; Barbour, A.; Gu, G.; Hill, J. P.; Tranquada, J. M.; Dean, M. P. M.; Wilkins, S. B.

    The nature of charge stripe order in the cuprates, and in particular whether the stripes are static or dynamic, is a key issue in understanding the relationship between stripes and superconductivity. In La2-xBaxCuO4 (LBCO) a low temperature structural distortion is widely believed to pin stripes into fixed, static domains, but such an assertion has never been directly verified. We performed resonant soft x-ray photon correlation spectroscopy (XPCS) to probe the charge order Bragg peak of 1/8 doped LBCO. At low temperatures, we observe time-independent x-ray speckle patterns persisting for more than three hours, proving the static nature of the stripes and we go on to discuss how stripe order melts with increasing temperature. Our results demonstrate that the combination of XPCS with diffraction limited light sources such as the National Synchrotron Light Source II can probe the dynamics of even subtle order parameters such as stripes in the cuprates. Work performed at Brookhaven National Laboratory was supported by the US Department of Energy, Division of Materials Science, under Contract No. DE-AC02-98CH10886. Use of the National Synchrotron Light Source II was supported under Contract No. DE-SC0012704.

  3. Increase in intracellular PGE2 induces apoptosis in Bax-expressing colon cancer cell

    NSAIDs exhibit protective properties towards some cancers, especially colon cancer. Yet, it is not clear how they play their protective role. PGE2 is generally shown as the only target of the NSAIDs anticancerous activity. However, PGE2 known targets become more and more manifold, considering both the molecular pathways involved and the target cells in the tumour. The role of PGE2 in tumour progression thus appears complex and multipurpose. To gain understanding into the role of PGE2 in colon cancer, we focused on the activity of PGE2 in apoptosis in colon cancer cell lines. We observed that an increase in intracellular PGE2 induced an apoptotic cell death, which was dependent on the expression of the proapoptotic protein Bax. This increase was induced by increasing PGE2 intracellular concentration, either by PGE2 microinjection or by the pharmacological inhibition of PGE2 exportation and enzymatic degradation. We present here a new sight onto PGE2 in colon cancer cells opening the way to a new prospective therapeutic strategy in cancer, alternative to NSAIDs

  4. Human ribosomal protein L9 is a Bax suppressor that promotes cell survival in yeast.

    Eid, Rawan; Sheibani, Sara; Gharib, Nada; Lapointe, Jason F; Horowitz, Avital; Vali, Hojatollah; Mandato, Craig A; Greenwood, Michael T

    2014-05-01

    The identification of a human ribosomal protein L9 (hRPL9) cDNA as a sequence capable of suppressing the lethal effects of heterologously expressed murine Bax in yeast led us to investigate its antiapoptotic potential. Using growth and viability assays, we show that yeast cells heterologously expressing hRPL9 are resistant to the growth inhibitory and lethal effects of exogenously supplied copper, indicating that it has pro-survival properties. To explore potential mechanisms, we used yeast mutants defective in all three types of programmed cell death (apoptosis, necrosis, and autophagy). The ability to retain pro-survival function in all the mutants suggests that hRPL9 may regulate a common pro-death process. In contrast, the yeast RPL9 orthologues, RPL9A and RPL9B, have opposite effects when overexpressed in yeast. In effect, instead of showing resistance to stress, RPL9A and RPL9B overexpressing cells show reduced cell growth. Further analysis indicates that the effects of overexpressed RPL9A and RPL9B are not in themselves lethal, instead, they serve to increase cell doubling time. Thus, yeast RPL9s are more representative of RPs whose extra-ribosomal function is similar to that of tumor suppressors. Taken together, our results demonstrate that RPL9 represents a species- and sequence-specific regulator of cell growth and survival. PMID:24305165

  5. The effect of octreotide and bromocriptine on expression of a pro-apoptotic Bax protein in rat prolactinoma.

    Jolanta Kunert-Radek

    2004-03-01

    Full Text Available It is well established that disruption of apoptosis may lead to tumor initiation, progression or metastasis. It is also well documented that many anticancer drugs induce apoptosis. In the earlier studies, the dopamine D2 receptor agonist bromocriptine (BC and somatostatin analog octreotide (OCT were found to inhibit the growth of the estrogen-induced rat prolactinoma. Our previous investigations, applying the TUNEL method showed the involvement of the pro-apoptotic effect in the action of BC, and to a lesser degree, in the action of OCT. The aim of the present study was to investigate whether the pro-apoptotic action of these drugs involves the increased expression of Bax--a member of Bcl-2 protein family which is known to play an important role in the regulation of apoptosis. Male four-week Fisher 344 rats were used in the experiment. Capsules containing diethylstilboestrol (DES were implanted subcutaneously. Six weeks after the implantation the rats were given OCT (2 x 25 microg/animal/24, BC (3 mg/kg b.w./24 h or OCT and BC at the above doses for 10 days. Bax expression was detected by immunohistochemistry. Prolactin (PRL in blood serum was measured by radioimmunoassay (RIA. It has been found that both OCT and BC, alone or in combination, significantly reduce the tumor weight. Both OCT and BC suppressed PRL levels, but the inhibitory effect of BC was stronger than that of OCT. It has been found that the treatment with OCT and BC, alone or in combination, causes a significant increase in Bax expression in the rat prolactinoma cells. Our findings indicate that anti-tumoral action of bromocriptine and to some extent the action of octreotide in the experimental rat prolactinoma is connected with the induction of apoptosis and is associated with increased Bax expression.

  6. BAX and Tumor Suppressor TRP53 Are Important in Regulating Mutagenesis in Spermatogenic Cells in Mice1

    Xu, Guogang; Vogel, Kristine S.; McMahan, C. Alex; Herbert, Damon C.; Walter, Christi A.

    2010-01-01

    During the first wave of spermatogenesis, and in response to ionizing radiation, elevated mutant frequencies are reduced to a low level by unidentified mechanisms. Apoptosis is occurring in the same time frame that the mutant frequency declines. We examined the role of apoptosis in regulating mutant frequency during spermatogenesis. Apoptosis and mutant frequencies were determined in spermatogenic cells obtained from Bax-null or Trp53-null mice. The results showed that spermatogenic lineage a...

  7. Quantitative Expression of BAG1, BAX and BCL-2 genes in Human Embryos with Different Fragmentation Grades Derived from ART

    Poopak Eftekhari Yazdi; Azam Piltan; Mohamadreza Baghaban Eslaminejad; Leila Karimian; Hamid Gourabi; Mojtaba Rezazadeh; Mehdi Totonchi

    2010-01-01

    Objective: The purpose of this study was to evaluate the quantitative expressions ofBAG1, BAX and BCL-2 in human embryos with different fragmentation grades as derivedfrom assisted reproduction technology (ART).Materials and Methods: Fragmented and normal human 8-cell embryos were scoredaccording to the degree of fragmentation with an inverted microscope and divided intofour grades (grade І: no or minimal fragmentation (

  8. Evaluation of Bax and Bcl-2 Proteins Expression in the Rat Hippocampus due to Childhood Febrile Seizure

    SAEEDI BORUJENI, Mohammad Javad; Hami, Javad; Haghir, Hossein; Rastin, Maryam; Sazegar, Ghasem

    2016-01-01

    Objective Simple Febrile Seizure (SFS) is the most common seizure disorder in childhood, and is frequently described as inoffensive disorder. Nevertheless, there is evidence suggesting the association between neonatal febrile seizures and hippocampal abnormalities in adulthood. This study was conducted at evaluating the hippocampal expression of pro-apoptotic Bax and anti-apoptotic Bcl-2 proteins following SFS induction in rat neonates. Materials & Methods Febrile seizure was modeled by hyper...

  9. p53's mitochondrial translocation and MOMP action is independent of Puma and Bax and severely disrupts mitochondrial membrane integrity

    Sonja Wolff; Susan Erster; Gustavo Palacios; Ute M Moll

    2008-01-01

    p53's apoptotic program consists of transcription-dependent and transcription-independent pathways. In the latter, physical interactions between mitochondrial p53 and anti-and pro-apoptotic members of the Bcl2 family of mitochondrial permeability regulators are central. Using isogenic cell systems with defined deficiencies, we characterize in detail how mitochondrial p53 contributes to mitochondrial permeabilization, to what extent its action depends on other key Bcl2 family members and define its release activity. We show that mitochondrial p53 is highly efficient in inducing the release of soluble and insoluble apoptogenic factors by severely disrupting outer and inner mitochondrial membrane integrity. This action is associated with wild-type p53-induced oligomerization of Bax, Bak and VDAC and the formation of a stress-induced endogenous complex between p53 and cyclophilin D, normally located at the inner membrane. Tumor-derived p53 mutants are deficient in activating the Bax/Bak lipid pore. These actions are independent of Puma and Bax. Importantly, the latter distinguishes the mitochondrial from the cytosolic p53 death pathway.

  10. A Structural Viral Mimic of Prosurvival Bcl-2: A Pivotal Role for Sequestering Proapoptotic Bax and Bak

    Kvansakul,M.; van Delft, M.; Lee, E.; Gulbis, J.; Fairlie, W.; Huang, D.; Colman, P.

    2007-01-01

    Many viruses express antiapoptotic proteins to counter host defense mechanisms that would otherwise trigger the rapid clearance of infected cells. For example, adenoviruses and some {gamma}-herpesviruses express homologs of prosurvival Bcl-2 to subvert the host's apoptotic machinery. Myxoma virus, a double-stranded DNA virus of the pox family, harbors antiapoptotic M11L, its virulence factor. Analysis of its three-dimensional structure reveals that despite lacking any primary sequence similarity to Bcl-2, it adopts a virtually identical protein fold. This allows it to associate with BH3 domains, especially those of Bax and Bak. We found that M11L acts primarily by sequestering Bax and Bak, thereby blocking the killing action of these essential cell-death mediators. These findings expand the family of protein sequences that act like Bcl-2 to block apoptosis and support the conclusion that the prosurvival action of these proteins critically depends on their ability to bind and antagonize Bax and/or Bak.

  11. Melatonin restores normal Bax and Bcl-2 protein expression in the subgranular zone of the dentate gyrus in pinealectomized rats

    Shengchang Zhang; Shuang Zhao; Lu Bai; Mingming Guan; Jielin Mo; Ling Lan

    2011-01-01

    In this study, we sought to elucidate the effects of melatonin on learning and memory as well as apoptosis and expression of the Bax or Bcl-2 proteins in the subgranular zone of the dentate gyrus in pinealectomized rats. Using the Morris water maze and the olfactory memory tests, we found that the average escape latency in pinealectomized rats was clearly increased compared with sham-operated rats. Moreover, the average escape latency in the melatonin-treated and pinealectomized rats was longer than that in the sham-operated rats and shorter than that in the pinealectomized and untreated rats. Immunohistochemistry and terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) showed that there were fewer Bax immunoreactive cells and TUNEL-positive (apoptotic) cells but more Bcl-2 immunoreactive cells in the melatonin-treated rats compared with the pinealectomized rats. The sham-operated rats showed numbers of these cells similar to the melatonin-treated rats. These experimental findings demonstrate that melatonin treatment may reduce abnormal apoptosis by promoting gene expression of Bax and suppressing gene expression of Bcl-2 in the subgranular zone of the dentate gyrus in pinealectomized rats. These effects appear to result in the inhibition of cellular apoptosis and the improvement of spatial learning and memory in pinealectomized rats.

  12. Schiff Base Metal Derivatives Enhance the Expression of HSP70 and Suppress BAX Proteins in Prevention of Acute Gastric Lesion

    Shahram Golbabapour

    2013-01-01

    Full Text Available Schiff base complexes have appeared to be promising in the treatment of different diseases and disorders and have drawn a lot of attention to their biological activities. This study was conducted to evaluate the regulatory effect of Schiff base metal derivatives on the expression of heat shock proteins (HSP 70 and BAX in protection against acute haemorrhagic gastric ulcer in rats. Rats were assigned to 6 groups of 6 rats: the normal control (Tween 20 5% v/v, 5 mL/kg, the positive control (Tween 20 5% v/v, 5 mL/kg, and four Schiff base derivative groups named Schiff_1, Schiff_2, Schiff_3, and Schiff_4 (25 mg/kg. After 1 h, all of the groups received ethanol 95% (5 mL/kg but the normal control received Tween 20 (Tween 20 5% v/v, 5 mL/kg. The animals were euthanized after 60 min and the stomachs were dissected for histology (H&E, immunohistochemistry, and western blot analysis against HSP70 and BAX proteins. The results showed that the Schiff base metal derivatives enhanced the expression of HSP70 and suppressed the expression of BAX proteins during their gastroprotection against ethanol-induced gastric lesion in rats.

  13. Melatonin promotes Bax sequestration to mitochondria reducing cell susceptibility to apoptosis via the lipoxygenase metabolite 5-hydroxyeicosatetraenoic acid

    Radogna, Flavia

    2015-03-01

    Extra-neurological functions of melatonin include control of the immune system and modulation of apoptosis. We previously showed that melatonin inhibits the intrinsic apoptotic pathway in leukocytes via stimulation of high affinity MT1/MT2 receptors, thereby promoting re-localization of the anti-apoptotic Bcl-2 protein to mitochondria. Here we show that Bcl-2 sequesters pro-apoptotic Bax into mitochondria in an inactive form after melatonin treatment, thus reducing cell propensity to apoptosis. Bax translocation and the anti-apoptotic effect of melatonin are strictly dependent on the presence of Bcl-2, and on the 5-lipoxygenase (5-LOX) metabolite 5-hydroxyeicosatetraenoic acid (5-HETE), which we have previously shown to be produced as a consequence of melatonin binding to its low affinity target calmodulin. Therefore, the anti-apoptotic effect of melatonin requires the simultaneous, independent interaction with high (MT1/MT2) and low (calmodulin) affinity targets, eliciting two independent signal transduction pathways converging into Bax sequestration and inactivation. MT1/MT2 vs. lipoxygenase pathways are activated by 10-9 vs. 10-5M melatonin, respectively; the anti-apoptotic effect of melatonin is achieved at 10-5M, but drops to 10-9M upon addition of exogenous 5-HETE, revealing that lipoxygenase activation is the rate-limiting pathway. Therefore, in areas of inflammation with increased 5-HETE levels, physiological nanomolar concentrations of melatonin may suffice to maintain leukocyte viability.

  14. Preparation and Photocatalytic Properties of Sr2−xBaxTa3O10−yNz Nanosheets

    Tatsumi Ishihara

    2013-01-01

    Full Text Available Sr2−xBaxTa3O10−yNz (x = 0.0, 0.5, 1.0 nanosheets were prepared by exfoliating layered perovskite compounds (CsSr2−xBaxTa3O10−yNz. The Sr1.5Ba0.5Ta3O9.7N0.2 nanosheet showed the highest photocatalytic activity for H2 production from the water/methanol system among the Sr2−xBaxTa3O9.7N0.2 nanosheets prepared. In addition, Rh-loaded Sr1.5Ba0.5Ta3O9.6N0.3 nanosheet showed the photocatalytic activity for oxygen and hydrogen production from water. The ratio of hydrogen to oxygen evolved was around two. These results indicate that the Rh-loaded Sr1.5Ba0.5Ta3O9.6N0.3 nanosheet is a potential catalyst for photocatalytic water splitting.

  15. Effects of systemic administration of ciliary neurotrophic factor on Bax and Bcl-2 proteins in the lumbar spinal cord of neonatal rats after sciatic nerve transection

    A.C.S. Rezende

    2008-11-01

    Full Text Available Ciliary neurotrophic factor (CNTF is a cytokine that plays a neuroprotective role in relation to axotomized motoneurons. We determined the effect of daily subcutaneous doses of CNTF (1.2 µg/g for 5 days; N = 13 or PBS (N = 13 on the levels of mRNA for Bcl-2 and Bax, as well as the expression and inter-association of Bcl-2 and Bax proteins, and the survival of motoneurons in the spinal cord lumbar enlargement of 2-day-old Wistar rats after sciatic nerve transection. Five days after transection, the effects were evaluated on histological and molecular levels using Nissl staining, immunoprecipitation, Western blot analysis, and reverse transcriptase-polymerase chain reaction. The motoneuron survival ratio, defined as the ratio between the number of motoneurons counted on the lesioned side vs those on the unlesioned side, was calculated. This ratio was 0.77 ± 0.02 for CNTF-treated rats vs 0.53 ± 0.02 for the PBS-treated controls (P < 0.001. Treatment with CNTF modified the level of mRNA, with the expression of Bax RNA decreasing 18% (with a consequent decrease in the level of Bax protein, while the expression of Bcl-2 RNA was increased 87%, although the level of Bcl-2 protein was unchanged. The amount of Bcl-2/Bax heterodimer increased 91% over that found in the PBS-treated controls. These data show, for the first time, that the neuroprotective effect of CNTF on neonatal rat axotomized motoneurons is associated with a reduction in free Bax, due to the inhibition of Bax expression, as well as increased Bcl-2/Bax heterodimerization. Thus, the neuroprotective action of the CNTF on axotomized motoneurons can be related to the inhibition of this apoptotic pathway.

  16. Safety of PEGylated recombinant human full‐length coagulation factor VIII (BAX 855) in the overall context of PEG and PEG conjugates

    Stidl, R.; Fuchs, S; BOSSARD, M; Siekmann, J.; Turecek, P. L.; Putz, M.

    2015-01-01

    Introduction BAX 855 is a PEGylated human full‐length recombinant factor VIII (rFVIII) based on licensed rFVIII (ADVATE). The applied PEGylation technology has been optimized to retain functionality of the FVIII molecule, improve its pharmacokinetic properties and allow less frequent injections while maintaining efficacy. Aim The aim of this study was to confirm that the excellent safety profile of ADVATE remains unchanged after PEGylation. Methods Non‐clinical safety studies with BAX 855 and...

  17. Early apoptosis and cell death induced by ATX-S10Na ( Ⅱ)-mediated photodynamic therapy are Bax- and p53-dependent in human colon cancer cells

    Makoto Mitsunaga; Akihito Tsubota; Kohichi Nariai; Yoshihisa Namiki; Makoto Sumi; Tetsuya Yoshikawa; Kiyotaka Fujise

    2007-01-01

    AIM: To investigate the roles of Bax and p53 proteins in photosensitivity of human colon cancer cells by using lysosome-localizing photosensitizer, ATX-S10Na (Ⅱ).METHODS: HCT116 human colon cancer cells and Bax-null or p53-null isogenic derivatives were irradiated with a diode laser. Early apoptosis and cell death in response to photodynamic therapy were determined by MTT assays, annexin V assays, transmission electron microscopy assays, caspase assays and western blotting.RESULTS: Induction of early apoptosis and cell death was Bax- and p53-dependent. Bax and p53 were required for caspase-dependent apoptosis. The levels of anti-apoptotic Bcl-2 family proteins, Bcl-2 and Bcl-XL,were decreased in Bax- and p53-independent manner.CONCLUSION: Our results indicate that early apoptosis and cell death of human colon cancer cells induced by photodynamic therapy with lysosome-localizingphotosensitizer ATX-S10Na (Ⅱ) are mediated by p53-Bax network and Iow levels of Bcl-2 and Bcl-XL proteins.Our results might help in formulating new therapeutic approaches in photedynamic therapy.

  18. In-house validation study of the DuPont Qualicon BAX system Q7 instrument with the BAX system PCR Assay for Salmonella (modification of AOAC Official Method 2003.09 and AOAC Research Institute Performance-Tested Method 100201).

    Tice, George; Andaloro, Bridget; White, H Kirk; Bolton, Lance; Wang, Siqun; Davis, Eugene; Wallace, Morgan

    2009-01-01

    In 2006, DuPont Qualicon introduced the BAX system Q7 instrument for use with its assays. To demonstrate the equivalence of the new and old instruments, a validation study was conducted using the BAX system PCR Assay for Salmonella, AOAC Official Method 2003.09, on three food types. The foods were simultaneously analyzed with the BAX system Q7 instrument and either the U.S. Food and Drug Administration Bacteriological Analytical Manual or the U.S. Department of Agriculture-Food Safety and Inspection Service Microbiology Laboratory Guidebook reference method for detecting Salmonella. Comparable performance between the BAX system and the reference methods was observed. Of the 75 paired samples analyzed, 39 samples were positive by both the BAX system and reference methods, and 36 samples were negative by both the BAX system and reference methods, demonstrating 100% correlation. Inclusivity and exclusivity for the BAX system Q7 instrument were also established by testing 50 Salmonella strains and 20 non-Salmonella isolates. All Salmonella strains returned positive results, and all non-Salmonella isolates returned a negative response. PMID:19610394

  19. Intermittent hypoxia attenuates ischemia/reperfusion induced apoptosis in cardiac myocytes via regulating Bcl-2/Bax expression

    2003-01-01

    Intermittent hypoxia has been shown to provide myocardial protection against ishemia/reperfusion-induced injury.Cardiac myocyte loss through apoptosis has been reported in ischemia/reperfusion injury. Our aim was to investigate whether intermittent hypoxia could attenuate ischemia/reperfusion-induced apoptosis in cardiac myocytes and its potential mechanisms. Adult male Sprague-Dawley rats were exposed to hypoxia simulated 5000 m in a hypobaric chamber for 6 h/day, lasting 42 days. Normoxia group rats were kept under normoxic conditions. Isolated perfused hearts from both groups were subjected to 30 min of global ischemia followed by 60 min reperfusion.Incidence of apoptosis in cardiac myocytes was determined by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) and DNA agarose gel electrophoresis. Expressions of apoptosis related proteins,Bax and Bcl-2, in cytosolic and membrane fraction were detected by Western Blotting. After ischemia/reperfusion,enhanced recovery of cardiac function was observed in intermittent hypoxia hearts compared with normoxia group.Ischemia/reperfusion-induced apoptosis, as evidenced by TUNEL-positive nuclei and DNA fragmentation, was significantly reduced in intermittent hypoxia group compared with normoxia group. After ischemia/reperfusion,expression of Bax in both cytosolic and membrane fractions was decreased in intermittent hypoxia hearts compared with normoxia group. Although ischemia/reperfusion did not induce changes in the level of Bcl-2 expression in cytosolic fraction between intermittent hypoxia and normoxia groups, the expression of Bcl-2 in membrane fraction was upregulated in intermittent hypoxia group compared with normoxia group. These results indicated that the cardioprotection of intermittent hypoxia against ischemia/reperfusion injury appears to be in part due to reduce myocardial apoptosis. Intermittent hypoxia attenuated ischemia/reperfusion-induced apoptosis via increasing the ratio of Bcl

  20. Involvement of P53 and Bax/Bad triggering apoptosis in thioacetamide-induced hepatic epithelial cells

    Li-Hsuen Chen; Chia-Yu Hsu; Ching-Feng Weng

    2006-01-01

    AIM: Thioacetamide (TAA) has been used in studying liver fibrosis and cirrhosis, however, the mechanisms of TAA-induced apoptosis in liver are still unclear. The hepatic epithelial cell line clone 9 was cultured and treated with TAA to investigate the causes of cell death. METHODS: The cell viability of TAA-induced clone 9 cells was determined using MTT assay. Total cellular GSH in TAA-induced clone 9 cells was measured using a slight modification of the Tietze assay. The activity of caspase 3 in TAA-induced clone 9 cells was monitored by the cleavage of DEVD-p-nitroanaline. TUNEL assay and flow cytometry were applied for the determination of DNA fragmentation and the proportion of apoptosis in TAAinduced clone 9 cells, respectively. The alterations of caspase 3, Bad, Bax and Phospho-P53 contents in TAAinduced clone 9 cells were measured by Western blot. RESULTS: The experimental data indicated that TAA caused rat hepatic epithelial cell line clone 9 cell death in a dose-and time-dependent manner; 60% of the cells died (MTT assay) within 24 h after 100 mg/L TAA was applied. Apoptotic cell percentage (TUNEL assay) and caspase 3 activities were highest after 100 mg/L TAA was added for 8 h. The release of GSH and the elevation in caspase content after TAA treatment resulted in clone 9 cell apoptosis via oxidative stress and a caspasedependent mechanism. The phospho-p53, Bax and Bad protein expressions in clone 9 cells were increased after TAA treatment.CONCLUSION: These results reveal that TAA activates p53, increases caspase 3, Bax and Bad protein contents,perhaps causing the release of cytochrome c from mitochondria and the disintegration of membranes, leading to apoptosis of cells.

  1. The protective role of Bax Inhibitor-1 against chronic mild stress through the inhibition of monoamine oxidase A

    Hwa-Young Lee; Geum-Hwa Lee; Anu Marahatta; Shun-Mei Lin; Mi-Rin Lee; Kyu Yun Jang; Kyung Min Kim; Hee Jae Lee; Jae-Won Lee; Tarique Rajasaheb Bagalkot; Young-Chul Chung; Yong-Chul Lee; Hyung-Ryong Kim; Han-Jung Chae

    2013-01-01

    The anti-apoptotic protein Bax inhibitor-1 (BI-1) is a regulator of apoptosis linked to endoplasmic reticulum (ER) stress. It has been hypothesized that BI-1 protects against neuron degenerative diseases. In this study, BI-1−/− mice showed increased vulnerability to chronic mild stress accompanied by alterations in the size and morphology of the hippocampi, enhanced ROS accumulation and an ER stress response compared with BI-1+/+ mice. BI-1−/− mice exposed to chronic mild stress showed signif...

  2. Retinoids cause apoptosis in pancreatic cancer cells via activation of RAR-γ and altered expression of Bcl-2/Bax

    Pettersson, F; Dalgleish, A G; Bissonnette, R P; Colston, K W

    2002-01-01

    All-trans-retinoic acid and 9-cis-retinoic acid have been reported to have inhibitory effects on pancreatic adenocarcinoma cells and we have shown that this is partly due to induction of apoptosis. In this study, the mechanisms whereby 9-cis-retinoic acid induces apoptosis in these cells were investigated. An involvement of the Bcl-2 family of proteins was shown, such that 9-cis-retinoic acid causes a decrease in the Bcl-2/Bax ratio. Overexpression of Bcl-2 also resulted in inhibition of apop...

  3. On the moment of inertia in deformed Ba-Xe nuclei as deduced from gamma-gamma energy correlation experiments

    The γ-rays following reactions induced by bombarding targets of 114,116,118,120,122Sn with 118 MeV 12C ions were investigated using six NaI(Tl) detectors in a two-dimensional coincidence arrangement. Experimental energy-correlation spectra were extracted from the original coincidence matrices. The energy-correlation spectra exhibit the features expected for rotational nuclei and were used to deduce information on the moment of inertia Isup((2)) = ΔI/Δω. The gross properties of the behaviour of Isup((2)) in the Ba-Xe region are discussed together with their interpretation within the cranked shell model (CSM). (orig.)

  4. Influences of HIF-lα on Bax/Bcl-2 and VEGF expressions in rats with spinal cord injury

    Chen, Mao-Hua; Ren, Qing-Xian; Yang, Wen-Fa; Chen, Xiang-Lin; Lu, Chuan; Sun, Jun

    2013-01-01

    Hypoxia-inducible factor 1-alpha (HIF-1α) is a subunit of HIF-l and thought to be able to protect hypoxic cells from apoptosis or necrosis under ischemic and anoxic conditions. This study aimed to investigated whether recombinant adenovirus vector over-expressing HIF-lα could affect apoptosis-related proteins (Bcl-2 and Bax) and vascular endothelial growth factor (VEGF) in a rat spinal cord injury (SCI) model. A total of 60 male SD rats were divided into 4 groups: Sham, Control, Ad-Blank and ...

  5. Molecular cloning, characterization and expression analysis of (B-cell lymphoma-2 associated X protein) Bax in the orange-spotted grouper (Epinephelus coioides) after the Vibrio alginolyticus challenge.

    Luo, Sheng-Wei; Wang, Wei-Na; Sun, Zuo-Ming; Xie, Fu-Xing; Kong, Jing-Rong; Liu, Yuan; Cheng, Chang-Hong

    2016-07-01

    Bax is a pro-apoptotic member of Bcl-2 like superfamily, playing an important role in regulating the apoptosis. In this study, the full-length Bax (EcBax) was obtained, containing a 5'UTR of 64 bp, an ORF of 579 bp and a 3'UTR of 1021 bp. The EcBax gene encoded a polypeptide of 192 amino acids with an estimated molecular mass of 21.55 KDa and a predicted isoelectric point (pI) of 6.75. The deduced amino acid sequence analysis showed that EcBax comprised the conserved residues and the characteristic domains known to the critical function of Bax. qRT-PCR analysis revealed that EcBax mRNA was broadly expressed in all of the examined tissues, while the highest expression level was observed in blood, followed by the expression in liver, gill, spleen, kidney, heart, muscle and intestine. A sharp increase of EcBax expression was observed in the vibrio challenge group by comparing with those in the control. Subcellular localization analysis revealed that EcBax was predominantly localized in the cytoplasm. EcBax exerted a regulatory role in modulating the mitochondrial membrane potential, promoting the cytochrome c release, and then activating the downstream caspase signaling. Moreover, the overexpression of EcBax can decrease the cell viability and antagonize NF-kB, AP-1, Stat3 promoter activity in Hela cells. These results indicate that EcBax containing the conserved domain of pro-apoptotic member of Bcl-2 family may disrupt the mammalian signaling and play a regulative role in the apoptotic process. PMID:26905633

  6. The Impact of Adenosine Fast Induction of Myocardial Arrest during CABG on Myocardial Expression of Apoptosis-Regulating Genes Bax and Bcl-2

    Ahmed Shalaby

    2009-01-01

    Full Text Available Background. We studied the effect of fast induction of cardiac arrest with denosine on myocardial bax and bcl-2 expression. Methods and Results. 40 elective CABG patients were allocated into two groups. The adenosine group (n=20 received 250 μg/kg adenosine into the aortic root followed by blood potassium cardioplegia. The control group received potassium cardioplegia in blood. Bcl-2 and bax were measured. Bax was reduced in the postoperative biopsies (1.38 versus 0.47, P=.002 in the control group. Bcl-2 showed a reducing tendency (0.14 versus 0.085, P=.07. After the adenosine treatment, the expression of both bax (0.52 versus 0.59, P=.4 and bcl-2 (0.104 versus 0.107, P=.4 remained unaltered after the operation. Conclusion. Open heart surgery is associated with rapid reduction in the expression of apoptosis regulating genes bax and bcl-2. Fast Adenosine induction abolished changes in their expression.

  7. Enhanced levels of the apoptotic BAX/BCL-2 ratio in children with acute lymphoblastic leukemia and high-risk features

    Maria Kaparou

    2013-01-01

    Full Text Available It has been suggested that leukemia is characterized by an impaired balance between the proliferation of blood cells and their capacity to undergo apoptosis. The aim of this study was to examine the expression of key molecules related to apoptosis (BCL-2, BAX, FAS, FAS-L in children with acute lymphoblastic leukemia (ALL. Measurement of BCL-2 and BAX mRNA was performed by quantitative real-time PCR, and membrane expression of FAS and FAS-L was assessed by flow cytometry in bone marrow mononuclear cells, both at diagnosis and at remission following induction chemotherapy. At diagnosis, increased levels of the apoptotic BAX/BCL-2 ratio were observed in children older than 10 years and with higher white blood cell counts. A DNA index < 1.16 was associated with increased BAX/BCL-2, both at diagnosis and at remission, and the del(9p chromosome abnormality with increased BAX/BCL-2 at remission. The expression of the apoptotic receptor FAS was significantly higher at remission compared to diagnosis, which might reflect enhanced sensitivity of the leukemic clone to apoptosis and response to treatment. Altogether, our results highlight the association of apoptosis-related genes with clinical and cytogenetic prognostic parameters in pediatric ALL. A better understanding of the mechanisms and regulation of apoptosis should enable the design of novel targeted therapies for these patients.

  8. Bcl-2, Bax, and c-Fos expression correlates to RPE cell apoptosis induced by UV-light and daunorubicin

    Liang, Y G; Jorgensen, A G; Kaestel, C G; Wiencke, A K; Lui, G M; la Cour, M H; Röpke, C H; Nissen, Mogens Holst

    2000-01-01

    PURPOSE. The aim of this study was to determine the role of Bcl-2, Bcl-X L, Bax, and c-Fos in regulation of apoptosis, induced by ultraviolet-light A (UV-A) and daunorubicin (DNR), in retinal pigment epithelium (RPE) cells grown on bovine extracellular matrix (ECM)-coated or uncoated plastic dishes....... METHODS. Apoptosis in confluent RPE cells cultured on ECM-coated or uncoated dishes was induced by UV-A or DNR. Apoptosis was detected by 7-amino-actinomycin D labeling followed by flow cytometry and by terminal deoxy-transferase mediated X-dUTP nick end labeling (TUNEL). Cellular expression of Bcl-2, Bcl......-X L, Bax, and c-Fos was determined by the use of antibodies and flow cytometry, Western blot analysis, and immunocytochemical staining. RESULTS. Both UV-A and DNR induce apoptosis in human RPE cells in vitro. Human fetal RPE cells grown on ECM-coated dishes were significantly more resistant to UV-A or...

  9. Effect of 147Pm β-irradiation on bcl-2 gene and bax gene expression in human leukemia cells

    Objective: To study the effect of exposure to a pure β radiator-147Pm on bcl-2 gene expression in human leukemia HL-60 cells. Methods: Immunohistochemical technique was used to identify the expression of bcl-2 and bax proteins in HL-60 cells, as well as RNA dot blot hybridization to show the expression of bcl-2 mRNA in HL-60 cells. Results: The expression of bcl-2 protein was as high as 88% in control human HL-60 cells. Down regulation of bcl-2 protein expression which lowered to 53% was noted after exposure to 147Pm. The expression of bax protein was low in control cells, and after irradiation it did not obviously change. An obvious down regulation of bcl-2 mRNA expression was found in irradiated HL-60 cells. With increasing time of 147Pm exposure, the down regulation of bcl-2 became more evident. Conclusion: The apoptotic action of 147Pm β-rays in human HL-60 cells is associated with the down regulation of the apoptosis-suppressing gene bcl-2

  10. ROS-mediated lipopolysaccharide-induced apoptosis in INS-1 cells by modulation of Bcl-2 and Bax.

    DU, S-C; Ge, Q-M; Lin, N; Dong, Y; Su, Q

    2012-01-01

    Overproduction of reactive oxygen species (ROS) or exhaustion of antioxidants may cause oxidative stress which is a major factor of defective insulin secretion and increases apoptosis of pancreatic β-cells in diabetes. So there comes a consideration of whether antioxidant strategies can be used to protect deterioration of the β-cells. In this study, we explored the mechanism of oxidative stress mediated lipopolysaccharide (LPS) induced apoptosis in insulin secreting (INS-1) cells from a rat pancreatic β-cell line. ROS was monitored by using intracellular ROS capture dihydroethidium (DHE) and dihydrorhodamine123 (DHR123). Apoptosis rate was measured by flow cytometry (FCM). The pro-apoptotic gene Bax and anti-apoptotic gene Bcl-2 were analysed by Western blot and RT-PCR. The results demonstrate that LPS-stimulated INS-1 cells manifest intensified intracellular fluorescence in both dose- and time- dependent manners. Apoptosis rate of LPS stimulated INS-1 cells is significantly increased by FCM, with a significant increase in Bax/Bcl-2 ratio revealed by Western blot and RT-PCR. Furthermore, α-lipoic acid (α-LA) inhibits LPS-induced apoptosis, but can not restore the function of glucose stimulated insulin secretion (GSIS) in INS-1 cells. PMID:22455982

  11. Expression of p53, Bax and Bcl-2 proteins in hepatocytes in non-alcoholic fatty liver disease

    Anatol Panasiuk; Janusz Dzieciol; Bozena Panasiuk; Danuta Prokopowicz

    2006-01-01

    AIM: To analyze the protein expression essential for apoptosis in liver steatosis.METHODS: The expression of proapoptotic proteinsp53, Bax, and antiapoptotic Bcl-2 in hepatocytes with steatosis (SH) and without steatosis (NSH) was evaluated in 84 patients at various stages of non-alcoholic fatty liver disease (NAFLD).RESULTS: Immunohistochemical staining of liver tissue showed the activation of p53 protein in SH and NSH with increased liver steatosis, diminished Bcl-2 and slightly decreased Bax protein. Positive correlation was found between the stage of liver steatosis with p53 expression in SH (r = 0.54, P < 0.01) and NSH (r = 0.49,P < 0.01).The antiapoptotic protein Bcl-2 was diminished together with the advancement of liver steatosis, especially in non-steatosed hepatocytes (r =0.43, P < 001).CONCLUSION: Apoptosis is one of the most important mechanisms leading to hepatocyte elimination in NAFLD. The intensification of inflammation in NAFLD induces proapoptotic protein p53 with the inhibition of antiapoptotic Bcl-2.

  12. Trimethyltin-induced apoptosis is associated with upregulation of inducible nitric oxide synthase and Bax in a hippocampal cell line

    Trimethyltin (TMT) produces selective neuronal degeneration in the central nervous system (CNS), in which the hippocampus is the most sensitive area. Since previous studies have been conducted in either non-neural cells or mixed primary cultures, an immortalized hippocampal neuronal cell line (HT-22 cell) was used to assess the mechanism and mode of death produced by TMT. The compound produced a time- and concentration-dependent apoptotic death that was caspase-mediated. Excessive generation of reactive oxygen species (ROS) and subsequent reduction of mitochondrial membrane potential (ΔΨm) were involved in the cytotoxicity. Scavenging of ROS by a free radical trapping agent or inhibition of the mitochondrial permeability transition (MPT) pore significantly reduced cell death. Additionally, TMT increased expression of inducible nitric oxide synthase (iNOS) by activation of the redox-sensitive transcription factor NFκB. Pharmacologic inhibition studies showed that the iNOS-mediated NO generation increased expression of Bax and then mitochondrial-mediated apoptosis. It was concluded that excessive ROS generation initiated the apoptotic cell death by upregulating iNOS followed by increased Bax expression which then led to loss of ΔΨm and caspase-executed cell death. This study is the first to report in a neuronal cell model that TMT stimulates induction of iNOS, which then increases cellular levels of reactive nitrogen species (RNS) to initiate apoptotic death

  13. Involvement of mitochondria in acetaminophen-induced apoptosis and hepatic injury Roles of cytochrome c, Bax, Bid, and caspases

    The role of apoptosis in acetaminophen (AAP)-induced hepatic injury was investigated. Six hours after AAP administration to BALB/c mice, a significant loss of hepatic mitochondrial cytochrome c was observed that was similar in extent to the loss observed after in vivo activation of CD95 by antibody treatment. AAP-induced loss of mitochondrial cytochrome c coincided with the appearance in the cytosol of a fragment corresponding to truncated Bid (tBid). At the same time, tBid became detectable in the mitochondrial fraction, and concomitantly, Bax was found translocated to mitochondria. However, AAP failed to activate the execution caspases 3 and 7 as evidenced by a lack of procaspase processing and the absence of an increase in caspase-3-like activity. In contrast, the administration of the pan-inhibitor of caspases, benzyloxycarbonyl-Val-Ala-DL-Asp-fluoromethylketone (but not its analogue benzyloxycarbonyl-Phe-Ala-fluoromethylketone) prevented the development of liver injury by AAP and the appearance of apoptotic parenchymal cells. This correlated with the inhibition of the processing of Bid to tBid. The caspase inhibitor failed to prevent both the redistribution of Bax to the mitochondria and the loss of cytochrome c. In conclusion, apoptosis is an important causal event in the initiation of the hepatic injury inflicted by AAP. However, as suggested by the lack of activation of the main execution caspases, apoptosis is not properly executed and degenerates into necrosis

  14. Apoptosis and the activity of ceramide, Bax and Bcl-2 in the lungs of neonatal rats exposed to limited and prolonged hyperoxia

    Bitar Fadi F

    2006-07-01

    Full Text Available Abstract Background The aim of the study is to examine the effect of limited and prolonged hyperoxia on neonatal rat lung. This is done by examining the morphologic changes of apoptosis, the expression of ceramide, an important mediator of apoptosis, the expression of inflammatory mediators represented by IL-1β and the expression of 2 proto-oncogenes that appear to modulate apoptosis (Bax and Bcl-2. Methods Newborn rats were placed in chambers containing room air or oxygen above 90% for 7 days. The rats were sacrificed at 3, 7 or 14 days and their lungs removed. Sections were fixed, subjected to TUNEL, Hoechst, and E-Cadherin Staining. Sections were also incubated with anti-Bcl-2 and anti-Bax antisera. Bcl-2 and Bax were quantitated by immunohistochemistry. Lipids were extracted, and ceramide measured through a modified diacylglycerol kinase assay. RT-PCR was utilized to assess IL-1β expression. Results TUNEL staining showed significant apoptosis in the hyperoxia-exposed lungs at 3 days only. Co-staining of the apoptotic cells with Hoechst, and E-Cadherin indicated that apoptotic cells were mainly epithelial cells. The expression of Bax and ceramide was significantly higher in the hyperoxia-exposed lungs at 3 and 14 days of age, but not at 7 days. Bcl-2 was significantly elevated in the hyperoxia-exposed lungs at 3 and 14 days. IL-1β expression was significantly increased at 14 days. Conclusion Exposure of neonatal rat lung to hyperoxia results in early apoptosis documented by TUNEL assay. The early rise in Bax and ceramide appears to overcome the anti-apoptotic activity of Bcl-2. Further exposure did not result in late apoptotic changes. This suggests that apoptotic response to hyperoxia is time sensitive. Prolonged hyperoxia results in acute lung injury and the shifting balance of ceramide, Bax and Bcl-2 may be related to the evolution of the inflammatory process.

  15. Determination of stoichiometry and concentration of trace elements in thin BaxSr1-xTiO3 perovskite layers.

    Becker, J S; Boulyga, S F

    2001-07-01

    This paper describes an analytical procedure for determining the stoichiometry of BaxSr1-xTiO3 perovskite layers using inductively coupled plasma mass spectrometry (ICP-MS). The analytical results of mass spectrometry measurements are compared to those of X-ray fluorescence analysis (XRF). The performance and the limits of solid-state mass spectrometry analytical methods for the surface analysis of thin BaxSr1-xTiO3 perovskite layers sputtered neutral mass spectrometry (SNMS)--are investigated and discussed. PMID:11496982

  16. Effects of Bisphenol A on apoptosis of spermatogenic cells and expression of Bax and Bcl-2 proteins in the Siberian frog Rana chensinensis

    LIN Sheng-Nan; Zhang, Yu-hui

    2008-01-01

    In order to investigate the effects of bisphenol A (BPA) on apoptosis and the expression levels of Bax and Bcl-2 protein in amphibian spermatogenic cells, we treated Rana chensinensis with concentrations of 10-7, 10-6, 10-5mol/L BPA respectively. Testes were removed after R.chensinensis had been treated for 3 , 5 or 7 days. Apoptotic cells were examined by TUNEL and Methyl Green-Pyronine. The expression levels of Bax and Bcl-2 protein in spermatogenic cells were detected by immunohistochemic...

  17. Curcuma purpurascens BI. rhizome accelerates rat excisional wound healing: involvement of Hsp70/Bax proteins, antioxidant defense, and angiogenesis activity

    Rouhollahi E

    2015-10-01

    Full Text Available Elham Rouhollahi,1 Soheil Zorofchian Moghadamtousi,2 Fatemeh Hajiaghaalipour,3 Maryam Zahedifard,2 Faezeh Tayeby,2 Khalijah Awang,4 Mahmood Ameen Abdulla,3 Zahurin Mohamed1 1Pharmacogenomics Laboratory, Department of Pharmacology, Faculty of Medicine, 2Institute of Biological Sciences, Faculty of Science, 3Department of Biomedical Science, Faculty of Medicine, 4Department of Chemistry, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia Purpose: Curcuma purpurascens BI. is a member of Zingiberaceae family. The purpose of this study is to investigate the wound healing properties of hexane extract of C. purpurascens rhizome (HECP against excisional wound healing in rats.Materials and methods: Twenty four rats were randomly divided into 4 groups: A negative control (blank placebo, acacia gum, B low dose of HECP, C high dose of HECP, and D positive control, with 6 rats in each group. Full-thickness incisions (approximately 2.00 cm were made on the neck area of each rat. Groups 1–4 were treated two-times a day for 20 days with blank placebo, HECP (100 mg/kg, HECP (200 mg/kg, and intrasite gel as a positive control, respectively. After 20 days, hematoxylin and eosin and Masson’s trichrome stainings were employed to investigate the histopathological alterations. Protein expressions of Bax and Hsp70 were examined in the wound tissues using immunohistochemistry analysis. In addition, levels of enzymatic antioxidants and malondialdehyde representing lipid peroxidation were measured in wound tissue homogenates.Results: Macroscopic evaluation of wounds showed conspicuous elevation in wound contraction after topical administration of HECP at both doses. Moreover, histopathological analysis revealed noteworthy reduction in the scar width correlated with the enhanced collagen content and fibroblast cells, accompanied by a reduction of inflammatory cells in the granulation tissues. At the molecular level, HECP facilitates wound-healing process

  18. Study of BaxSr1-xTiO3 thin films using transverse-field Ising model

    Tao Yong-Mei; Jiang Qing

    2004-01-01

    In this paper, the effects of doping on the thermodynamic properties of BaxSr1-xTiO3 (BST) thin film are investigated, based on the transverse-field Ising model (TIM) within the framework of mean field theory. We apply the double-peak distribution model of related parameters to mimic doping. The lattice expansion arising from doping with large Ba2+ was also taken into account. We concentrate on the doping concentration dependence of peak temperature (Tm), spontaneous polarization and dielectric susceptibility. It is found that the doping concentration has great influence on the dielectric properties and phase transition properties of BST thin films. We also discuss the quantum effect arising from doping.

  19. Lattice instabilities, isotope effect, and high-Tc superconductivity in La2-xBaxCuO4

    Unlike La2-xSrxCuO4, La2-xBaxCuO4, x∼xcr=0.12, undergoes a structural change to a low-temperature tetragonal (LTT) phase below 60 K, and this transformation is known to suppress Tc drastically. We present self-consistent local-density-functional calculations for this system that indicate the CuO6 octahedrons are unstable to an X-point tilt in any direction, thereby leading to strongly anharmonic X-point vibrations. The LTT phase is most favored energetically, consistent with experiment. The effect of the LTT distortion on the electronic structure accounts for the suppression of Tc in the LTT phase, and for the enhanced isotope effect at x=xcr

  20. Dietary anthocyanins protect endothelial cells against peroxynitrite-induced mitochondrial apoptosis pathway and Bax nuclear translocation: an in vitro approach.

    Paixão, Joana; Dinis, Teresa C P; Almeida, Leonor M

    2011-10-01

    Anthocyanins have received increasing attention because of their relatively high intake in humans and wide range of potential health-promoting effects, including anti-atherogenic properties. Evidences support their vascular protective effects but the involved molecular mechanisms have not been well clarified. The endothelium seems to have a central role in atherogenesis and apoptosis is emerging as a crucial event in this disease progression. Following our previous work on the biochemical pathways underlying peroxynitrite-triggered apoptosis in endothelial cells, here we investigated potential mechanisms responsible for the cytoprotective actions of three common anthocyanins, namely cyanidin- delphinidin- and pelargonidin-3-glucoside, against this process. Beyond their antioxidant properties, all these flavonoids, possessing either catecholic or monophenolic structures, were able to counteract peroxynitrite-induced apoptotic effects in endothelial cells through the inhibition of several crucial signaling cascades. Actually, pre-incubation of cells with 25 μM anthocyanins prevented them from peroxynitrite-mediated apoptosis, which was evaluated by the loss of mitochondrial membrane potential, caspases-9 and-3 activation, the increase in cytoplasmatic Bax levels and the inactivation of the PI3 K/Akt pathway. Moreover, they counteracted the translocation of Bax into the nucleus, as observed by immunocytochemistry and immunoblot, an event shown for the first time in endothelial cells apoptotic process. Such cellular actions could not be inferred from their in vitro antioxidant properties. These results suggest a potential role of dietary anthocyanins in the modulation of several apoptotic signaling pathways triggered by peroxynitrite in endothelial cells, supporting mechanistically their health benefits in the context of prevention of endothelial dysfunction and, ultimately, of atherosclerosis. PMID:21785847

  1. Quantitative Expression of BAG1, BAX and BCL-2 genes in Human Embryos with Different Fragmentation Grades Derived from ART

    Poopak Eftekhari Yazdi

    2010-01-01

    Full Text Available Objective: The purpose of this study was to evaluate the quantitative expressions ofBAG1, BAX and BCL-2 in human embryos with different fragmentation grades as derivedfrom assisted reproduction technology (ART.Materials and Methods: Fragmented and normal human 8-cell embryos were scoredaccording to the degree of fragmentation with an inverted microscope and divided intofour grades (grade І: no or minimal fragmentation (25% fragmentation and grade ІV: apoptotic inducedembryos with actinomycin D. In this study, TUNEL labeling was initially used todetect apoptosis, and then revers transcription polymerase chain reaction (RT-PCR andquantitative PCR were used to define the quantitative expressions of experimental genesin human embryos with different fragmentation grades.Results: The results of TUNEL labeling showed that embryos with higher fragmentationhad a high number of apoptotic bodies. The results of RT-PCR and q-PCR analysesshowed a significantly decreased amount of BAGI transcript expression from group I togroup IV. The highest expression of BAX gene was observed in group II, however, thetranscript of BCL-2 gene was not observed in any of the experimental groups. The effectof actinomycin D on transcript expression amounts of experimental genes in apoptoticinduced embryos (group IV compared to control embryos (group I showed a significantdecrease.Conclusion: mRNA expression of BAG1 gene can be used as a good marker to detectapoptosis in human embryos. However, the transcript of BCL-2 gene does not play a rolein the detection of apoptosis in human embryos at the 8-cell stage.

  2. Ganoderma lucidum spore powder modulates Bcl-2 and Bax expression in the hippocampus and cerebral cortex, and improves learning and memory in pentylenetetrazole-kindled rats

    Shuang Zhao; Shengchang Zhang; Shuqiu Wang

    2011-01-01

    We studied the effects of Ganoderma lucidum spore powder on Bax and Bcl-2 expression and neuronal apoptosis in pentylenetetrazole-kindled epileptic rats. Sixty adult rats were randomly divided into a control group, an epileptic group (kindled) and three medication groups ( 150, 300,450 mg/kg given to kindled rats). Bax and Bcl-2 immunohistochemistry and TUNEL labeling show ed that the number of Bax- and TUNEL-positive cells in the hippocampus and cerebral cortex decreased significantly in the high-dose medication group, while the number of Bcl-2immunoreactive cells increased. The Morris water maze test showed that high-dose treatment significantly shortened escape latency and increased spatial probe trial performance. Our findings indicate that a high dose of Ganoderma lucidum spore powder upregulates the expressionof antiapoptotic Bcl-2 protein in the hippocampus and cerebral cortex, inhibits proapoptotic Bax expression, and decreases seizure-induced neuronal apoptosis. Further,Ganoderma lucidum appears to protect against epilepsy-related learning and memory impairments.

  3. BaxSr1−xTi1.02O3 metal–insulator–metal capacitors on planarized alumina substrates

    Tiggelman, M.P.J.; Reimann, K.; Klee, M.; Mauczok, R.; Keur, W.; Hueting, R.J.E.

    2010-01-01

    Nanocrystalline barium strontium titanate (BaxSr1−xTi1.02O3) thin films with a barium content of x=0.8, 0.9 and 1 have been fabricated in a metal–insulator–metal configuration on glass-planarized alumina substrates. Cost-effective processing measures have been utilized by using poly-crystalline alum

  4. The trade-off between tuning ratio and quality factor of BaxSr1-xTiO3 MIM capacitors on alumina substrates

    Tiggelman, M.P.J.; Reimann, K.; Liu, J.; Klee, M.; Mauczok, R.; Keur, W.; Schmitz, J.; Hueting, R.J.E.

    2008-01-01

    Barium strontium titanate with different compositions is deposited using wet-chemical processing on a glass planarization layer, on top of alumina substrates. Three samples were fabricated with BaxSr1-xTiO3 (BST) with the barium content x varying between 0.8 and 1. The poly-crystalline films are 530

  5. Effect of compound preparation Tongqiao Jiannao capsules on neural cell apoptosis and Bcl-2 and Bax protein levels in a rat model of brain ischemia/reperfusion injury

    Rui Wang; Guanglai Li; Wei Wang; Huanying Li

    2008-01-01

    BACKGROUND: Pharmacological studies have demonstrated that compound preparation Tongqiao Jiannao capsules composed of Zexie, Baizhu, Honghua, Danshen, and Shexiang can supplement qi,activate blood circulation, relieve blood stasis, induce resuscitation for alleviating pain, relieve pain, anddilate blood vessels.OBJECTIVE: To observe the effects of Tongqiao Jiannao capsules on the levels of the anti-apoptotic protein Bcl-2 and the pro-apoptotic protein Bax, and verify the mechanism of action.DESIGN, TIME AND SETTING: Randomized, controlled animal experiment, performed in the Laboratory of Biochemistry and Molecular Biology, Shanxi Medical University between June 2001 and December 2002.MATERIALS: The right middle cerebral arteries of 24 healthy adult Sprague Dawley rats were occluded by the suture method. The primary Chinese herbal medicinal ingredients of Tongqiao Jiannao capsules are Zexie. Baizhu, Honghua, Danshen, and Shexiang, which were purchased from Shanxi Provincial Medicinal Material Company, China, and prepared into condensed granules in the Room for Chinese Herbal Medicine Preparation, Second Hospital, Shanxi Medical University. Bcl-2 and Bax immunohistochemical staining kits, a 3,3-diaminobenzidine(DAB) kit, and an in situ apoptosis detection kit were purchased from Wuhan Boster Bioengineering Co., Ltd., China.METHODS: Twenty-four rats were randomly and evenly divided into three groups: (1) sham-operated rats in which sutures were inserted and immediately pulled out; (2) Tongqiao Jiannao capsule-treated rats that were intragastrically administered 6.5 g/kg/d Tongqiao Jiannao capsule preparation for seven successive days prior to middle cerebral artery occlusion (MCAO); and (3) MCAO rats without any other treatments.MAIN OUTCOME MEASURES: The levels of neural cell apoptosis and Bcl-2 and Bax proteins at 24 hours post-surgery.RESULTS: In the MCAO group, the numbers of apoptotic cells and Bax-positive cells were significantly increased, while the numbers of

  6. Photobiomodulation on Bax and Bcl-2 Proteins and SIRT1/PGC-1α Axis mRNA Expression Levels of Aging Rat Skeletal Muscle

    Fang-Hui Li

    2014-01-01

    Full Text Available Objective. This study aimed to analyze the effects of low level laser irradiation (LLLI on Bax and IGF-1 and Bcl-2 protein contents and SIRT1/PGC-1α axis mRNA expression levels to prevent sarcopenia in aged rats. Material and Methods. Twenty female Sprague Dawley rats (18 months old were randomly divided into two groups (n=10 per group: control (CON and LLLI groups. The gallium-aluminum-arsenium (GaAlAs laser irradiation at 810 nm was used in the single point contact mode (3.75 J/cm2; 0.4 cm2; 125 mW/cm2; 30 s. Bax, Bcl-2, and IGF-1 proteins and SIRT1/PGC-1α axis mRNA expression were assessed 24 h after LLLI on gastrocnemius in aged rat. Results. Gastrocnemius muscle weights, gastrocnemius mass/body mass, Bcl-2/BAX ratio, Bcl-2 protein, IGF-1 protein, and the mRNA contents in SIRT1, PGC-1α, NRF1, TMF, and SOD2 were significantly (P<0.05 increased by LLLI compared to CON group without LLLI. However, levels of BAX protein and caspase 3 mRNA were significantly attenuated by LLLI compared to CON group (P<0.05. Conclusion. LLLI at 810 nm inhibits sarcopenia associated with upregulation of Bcl-2/BAX ratio and IGF-1 and SIRT1/PGC-1α axis mRNA expression in aged rats. This indicates that LLLI has potential to decrease progression of myocyte apoptosis in sarcopenic muscles.

  7. Identification of Novel Pepper Genes Involved in Bax- or INF1-Mediated Cell Death Responses by High-Throughput Virus-Induced Gene Silencing

    Jeong Hee Lee

    2013-11-01

    Full Text Available Hot pepper is one of the economically important crops in Asia. A large number of gene sequences, including expressed sequence tag (EST and genomic sequences are publicly available. However, it is still a daunting task to determine gene function due to difficulties in genetic modification of a pepper plants. Here, we show the application of the virus-induced gene silencing (VIGS repression for the study of 459 pepper ESTs selected as non-host pathogen-induced cell death responsive genes from pepper microarray experiments in Nicotiana benthamiana. Developmental abnormalities in N. benthamiana plants are observed in the 32 (7% pepper ESTs-silenced plants. Aberrant morphological phenotypes largely comprised of three groups: stunted, abnormal leaf, and dead. In addition, by employing the combination of VIGS and Agrobacterium-mediated transient assays, we identified novel pepper ESTs that involved in Bax or INF1-mediated cell death responses. Silencing of seven pepper ESTs homologs suppressed Bax or INF1-induced cell death, five of which suppressed both cell death responses in N. benthamiana. The genes represented by these five ESTs encode putative proteins with functions in endoplasmic reticulum (ER stress and lipid signaling. The genes represented by the other two pepper ESTs showing only Bax-mediated cell death inhibition encode a CCCH-type zinc finger protein containing an ankyrin-repeat domain and a probable calcium-binding protein, CML30-like. Taken together, we effectively isolated novel pepper clones that are involved in hypersensitive response (HR-like cell death using VIGS, and identified silenced clones that have different responses to Bax and INF1 exposure, indicating separate signaling pathways for Bax- and INF1-mediated cell death.

  8. Runx3 Expression Inhibits Proliferation and Distinctly Alters mRNA Expression of Bax in AGS and A549 Cancer Cells

    Torshabi, Maryam; Faramarzi, Mohammad Ali; Tabatabaei Yazdi, Mojtaba; Ostad, Seyyed Naser; Gharemani, Mohammad Hosein

    2011-01-01

    Runx3, a member of Runt-related transcription factor (Runx) proteins with tumor suppressor effect, is a tissue–restricted and cancer related transcription factor that regulate cell proliferation and growth, as well as differentiation. In the present study, exogenous Run3 was transiently expressed in AGS (human gastric adenocarcinoma), with undetectable Runx3 protein and in A549 (human lung carcinoma) with low levels of endogenous Runx3 protein. The GFP tagged Runx3 was transfected into AGS and A549 cells using fugene6 and PolyFect and Runx3 expression was confirmed by fluorescent microscopy and RT-PCR. The effect of Runx3 transfection on cell proliferation was determined by MTT assay and the results were confirmed by the trypan blue dye exclusion method. The effect of Runx3 expression on mRNA expression of BCL2-associated X protein (Bax) was evaluated using RT-PCR. In AGS and A549 cells, Runx3 expression inhibited cell proliferation (p < 0.01). The growth inhibition was less in A549 cells. We show that Runx3 expression increases Bax mRNA expression in AGS cells when compared with control (p < 0.05), but no significant differences in mRNA expression was observed in both examined cells. Runx3 expression has antiproliferative effect in AGS cell perhaps via increase in expression of Bax. The effect of Runx3 on A549 cells’ viability which has endogenous level of Runx3 is not related to Bax. These findings implicate a complex regulation by Runx3 in inhibition of cell proliferation utilizing Bax. PMID:24250365

  9. Melatonin may play a role in modulation of bax and bcl-2 expression levels to protect rat peripheral blood lymphocytes from gamma irradiation-induced apoptosis

    The close relationship between free radicals effects and apoptosis process has been proved. Melatonin has been reported as a direct free radical scavenger. We investigated the capability of melatonin in the modification of radiation-induced apoptosis and apoptosis-associated upstream regulators expression in rat peripheral blood lymphocytes. Rats were irradiated with a single whole body Cobalt 60-gamma radiation dose of 8 Gy at a dose rate of 101 cGy/min with or without melatonin pretreatments at different concentrations of 10 and 100 mg/kg body weight. The rats were divided into eight groups of control, irradiation-only, vehicle-only, vehicle plus irradiation, 10 mg/kg melatonin alone, 10 mg/kg melatonin plus irradiation, 100 mg/kg melatonin alone and 100 mg/kg melatonin plus irradiation. Rats were given an intraperitoneal (IP) injection of melatonin or the same volume of vehicle alone 1 h prior to irradiation. Blood samples were taken 4, 24, 48 and 72 h after irradiation for evaluation of flow cytometric analysis of apoptotic lymphocytes using Annexin V/PI assay and measurement of bax and bcl-2 expression using quantitative real-time PCR (RT2qPCR). Irradiation-only and vehicle plus irradiation showed an increase in the percentage of apoptotic lymphocytes significantly different from control group (P < 0.01), while melatonin pretreatments in a dose-dependent manner reduced it as compared with the irradiation-only and vehicle plus irradiation groups (P < 0.01) in all time points. This reduced apoptosis by melatonin was related to the downregulation of bax, upregulation of bcl-2, and therefore reduction of bax/bcl-2 ratio. Our results suggest that melatonin in these doses may provide modulation of bax and bcl-2 expression as well as bax/bcl-2 ratio to protect rat peripheral blood lymphocytes from gamma irradiation-induced apoptosis.

  10. Melatonin may play a role in modulation of bax and bcl-2 expression levels to protect rat peripheral blood lymphocytes from gamma irradiation-induced apoptosis

    Mohseni, Mehran [Department of Radiology and Medical Physics, Faculty of Paramedicine, Kashan University of Medical Sciences, Kashan (Iran, Islamic Republic of); Mihandoost, Ehsan, E-mail: mihandoost.e@gmail.com [Department of Medical Radiation Engineering, Science and Research Branch, Islamic Azad University, Tehran (Iran, Islamic Republic of); Shirazi, Alireza [Department of Medical Physics and Biomedical Engineering, Faculty of Medicine, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Sepehrizadeh, Zargham [Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Bazzaz, Javad Tavakkoly [Department of Medical Genetics, Faculty of Medicine, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Ghazi-khansari, Mahmoud [Department of Pharmacology, Faculty of Medicine, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of)

    2012-10-15

    The close relationship between free radicals effects and apoptosis process has been proved. Melatonin has been reported as a direct free radical scavenger. We investigated the capability of melatonin in the modification of radiation-induced apoptosis and apoptosis-associated upstream regulators expression in rat peripheral blood lymphocytes. Rats were irradiated with a single whole body Cobalt 60-gamma radiation dose of 8 Gy at a dose rate of 101 cGy/min with or without melatonin pretreatments at different concentrations of 10 and 100 mg/kg body weight. The rats were divided into eight groups of control, irradiation-only, vehicle-only, vehicle plus irradiation, 10 mg/kg melatonin alone, 10 mg/kg melatonin plus irradiation, 100 mg/kg melatonin alone and 100 mg/kg melatonin plus irradiation. Rats were given an intraperitoneal (IP) injection of melatonin or the same volume of vehicle alone 1 h prior to irradiation. Blood samples were taken 4, 24, 48 and 72 h after irradiation for evaluation of flow cytometric analysis of apoptotic lymphocytes using Annexin V/PI assay and measurement of bax and bcl-2 expression using quantitative real-time PCR (RT{sup 2}qPCR). Irradiation-only and vehicle plus irradiation showed an increase in the percentage of apoptotic lymphocytes significantly different from control group (P < 0.01), while melatonin pretreatments in a dose-dependent manner reduced it as compared with the irradiation-only and vehicle plus irradiation groups (P < 0.01) in all time points. This reduced apoptosis by melatonin was related to the downregulation of bax, upregulation of bcl-2, and therefore reduction of bax/bcl-2 ratio. Our results suggest that melatonin in these doses may provide modulation of bax and bcl-2 expression as well as bax/bcl-2 ratio to protect rat peripheral blood lymphocytes from gamma irradiation-induced apoptosis.

  11. BaxSr1−xTiO3 thin films prepared by OTS monomolecular film reverse induction and liquid phase self-assembly method

    Highlights: • OTS-SAMs were prepared on the substrate by self-assembled monomolecular technique. • After UV-light irradiation, OTS-SAMs became hydrophilic monolayers in 1 nm thickness. • Ferroelectric BaxSr1−xTiO3 film can be prepared on substrate by the reverse LPD-SAMs. -- Abstract: Octadecyl-trichlorosilane self-assembled monolayers (OTS-SAMs) were prepared on the ITO glass substrate surface by self-assembled monomolecular film technique and BaxSr1−xTiO3 thin film was prepared by the reverse induction and adsorption on the functionalized substrate surface. The morphologies of OTS-SAMs before and after UV-light irradiation, the variation of the contact angles, and the microstructure and electrical property of BaxSr1−xTiO3 thin film were investigated. The results show that after UV-light irradiation, the hydrophilic film of OTS-SAMs with the thickness of 1 nm and the contact angle of 5° can be used to prepare the homogeneous BaxSr1−xTiO3 thin film by the reverse induction method. As the increase of Ba content, the diffraction peaks corresponding to SrTiO3 crystal plane is shifted to small angles. The grains of the thin film are decreased as the increase of Ba doping content. When Sr/Ba is between 9/1 and 8/2, the dielectric constant of BaxSr1−xTiO3 thin film is higher while the dielectric loss is smaller. At 10 kHz, the dielectric constant of Ba0.2Sr0.8TiO3 thin film is 880 and the dielectric loss is 0.04. The remnant polarization of BaxSr1−xTiO3 thin film prepared by the reverse induction and adsorption and liquid phase self-assembly method is 0.64 μC/cm2 and the coercivity is 13.84 kV/cm

  12. Differential Effects of Ethanol on c-Jun N-Terminal Kinase, 14-3-3 Proteins, and Bax in Postnatal Day 4 and Postnatal Day 7 Rat Cerebellum

    Heaton, Marieta Barrow; Paiva, Michael; Kubovic, Stacey; Kotler, Alexandra; Rogozinski, Jonathan; Swanson, Eric; Madorsky, Vladimir; Posados, Michelle

    2011-01-01

    These studies investigated ethanol effects on upstream cellular elements and interactions which contribute to Bax-related apoptosis in neonatal rat cerebellum at ages of peak ethanol sensitivity (postnatal day 4 [P4]), compared to later ages of relative resistance (P7). Analyses were made of basal levels of the pro-apoptotic c-jun N-termimal kinase (JNK), Bax, and the 14-3-3 anchoring proteins, as well as the responsiveness of these substances to ethanol at P4 versus P7. Dimerization of Bax w...

  13. Role of the lattice dynamics in La2-xBaxCuO4 superconductor based on DFT method

    A Tavana

    2010-09-01

    Full Text Available Electron-phonon coupling parameters are calculated for La2-x BaxCuO4 cuprate superconductor in a wide range of dopings, from undoped to overdoped compounds. In this study we aim to study the quality of such calculations based on DFT method so, the results of σ GGA+U electronic structure calculations are also investigated. The obtained value for electron-phonon coupling is in the same order of previous calculations but, the value obtained for the Hubbard U parameter shows that, such methods are poor in the estimation of electronic correlations to decide about the role of phonons in these compounds based on their results. Moreover, existence of several structural phase transitions with temperature and doping, lead to larger error in these calculations. Based on the calculated phonon dispersions, structural phase transitions can be resulted which shows the ability of DFT in the study of structural properties and the weakness of the strongly correlations in this properties.

  14. Grape Seed Procyanidin Extract Improves Insulin Production but Enhances Bax Protein Expression in Cafeteria-Treated Male Rats

    Lídia Cedó

    2013-01-01

    Full Text Available In a previous study, the administration of a grape seed procyanidin extract (GSPE in female Wistar rats improved insulin resistance, reduced insulin production, and modulated apoptosis biomarkers in the pancreas. Considering that pharmacokinetic and pharmacodynamic parameters in females are different from these parameters in males, the aim of the present study was to evaluate the effects of GSPE on male Wistar cafeteria-induced obese rats. The results have confirmed that the cafeteria model is a robust model mimicking a prediabetic state, as these rats display insulin resistance, increased insulin synthesis and secretion, and increased apoptosis in the pancreas. In addition, GSPE treatment (25 mg/kg of GSPE for 21 days in male rats improves insulin resistance and counteracts the cafeteria-induced effects on insulin synthesis. However, the administration of the extract enhances the cafeteria-induced increase in Bax protein levels, suggesting increased apoptosis. This result contradicts previous results from cafeteria-fed female rats, in which GSPE seemed to counteract the increased apoptosis induced by the cafeteria diet.

  15. Plumbagin reduces chronic lymphocytic leukemia cell survival by downregulation of Bcl-2 but upregulation of the Bax protein level.

    Fu, Chunling; Gong, Yanqing; Shi, Xuanxuan; Sun, Zengtian; Niu, Mingshan; Sang, Wei; Xu, Linyan; Zhu, Feng; Wang, Ying; Xu, Kailin

    2016-09-01

    Chronic lymphocytic leukemia (CLL) is the most common leukemia in Western countries, and mainly originates from an accumulation of abnormal B cells caused by the dysregulation of cell proliferation and apoptosis rates. The aberration of apoptosis-related genes in CLL cells results in defective apoptosis of CLL cells in response to traditional therapeutic medicine. Plumbagin (5-hydroxy-2-methyl-1, 4-naphthoquinone), a natural compound from Plumbago zeylinica, has been shown to exhibit pro-apoptotic activities in tumor cells. In the present study, we report that plumbagin effectively inhibited CLL cell viability with a lower dose compared to fludarabine, and inhibited cell proliferation in a dose-dependent manner. In addition, plumbagin promoted accumulation of MEC-1 cells in the S phase, and blocked cell cycle transition of HG3 cells from G0/G1 to S phase. Molecularly, plumbagin markedly induced CLL cell apoptosis through reduction of Bcl-2, but through an increase in the Bax protein level. These results suggest that plumbagin may be considered as a potential anticancer agent for CLL therapy. PMID:27461100

  16. The N-terminus and alpha-5, alpha-6 helices of the pro-apoptotic protein Bax, modulate functional interactions with the anti-apoptotic protein Bcl-xL

    Sowdhamini R

    2007-05-01

    Full Text Available Abstract Background Bcl-2 family proteins are key regulators of mitochondrial integrity and comprise both pro- and anti-apoptotic proteins. Bax a pro-apoptotic member localizes as monomers in the cytosol of healthy cells and accumulates as oligomers in mitochondria of apoptotic cells. The Bcl-2 homology-3 (BH3 domain regulates interactions within the family, but regions other than BH3 are also critical for Bax function. Thus, the N-terminus has been variously implicated in targeting to mitochondria, interactions with BH3-only proteins as well as conformational changes linked to Bax activation. The transmembrane (TM domains (α5-α6 helices in the core and α9 helix in the C-terminus in Bax are implicated in localization to mitochondria and triggering cytotoxicity. Here we have investigated N-terminus modulation of TM function in the context of regulation by the anti-apoptotic protein Bcl-xL. Results Deletion of 29 amino acids in the Bax N-terminus (Bax 30–192 caused constitutive accumulation at mitochondria and triggered high levels of cytotoxicity, not inhibited by Bcl-xL. Removal of the TM domains (Bax 30–105 abrogated mitochondrial localization but resulted in Bcl-xL regulated activation of endogenous Bax and Bax-Bak dependent apoptosis. Inclusion of the α5-α6 helices/TMI domain (Bax 30–146 phenocopied Bax 30–192 as it restored mitochondrial localization, Bcl-xL independent cytotoxicity and was not dependent on endogenous Bax-Bak. Inhibition of function and localization by Bcl-xL was restored in Bax 1–146, which included the TM1 domain. Regardless of regulation by Bcl-xL, all N-terminal deleted constructs immunoprecipitated Bcl-xLand converged on caspase-9 dependent apoptosis consistent with mitochondrial involvement in the apoptotic cascade. Sub-optimal sequence alignments of Bax and Bcl-xL indicated a sequence similarity between the α5–α6 helices of Bax and Bcl-xL. Alanine substitutions of three residues (T14A-S15A-S16A in

  17. Ab-initio study of structural, elastic, electronic and thermodynamic properties of BaxSr1−xS ternary alloys

    Chelli S.

    2015-12-01

    Full Text Available The structural, elastic, electronic and thermodynamic properties of BaxSr1−xS ternary alloys have been investigated using the full-potential (linearized augmented plane wave method. The ground state properties, such as lattice constant, bulk modulus and elastic constants, are in good agreement with numerous experimental and theoretical data. The dependence of the lattice parameters, bulk modulus and band gap on the composition x was analyzed. Deviation of the lattice constant from Vegard’s law and the bulk modulus from linear concentration dependence (LCD was observed. The microscopic origins of the gap bowing were explained by using the approach of Zunger et al. The thermodynamic stability of BaxSr1−xS alloy was investigated by calculating the excess enthalpy of mixing, ΔHm and the calculated phase diagram showed a broad miscibility gap with a critical temperature.

  18. Injury of mitochondria and the expressions of fas and bax mRNA in the hip-pus of epileptic rats of different latency

    Shuhai Tang; Jianying Sun; Xiaojun Pan; Li Zhang

    2006-01-01

    BACKGROUND: It has been confirmed that Fas and Bax respectively mediate the exogenous and endogenous pathways of neuronal apoptosis, and then mediate the neuronal injury after status epilepticus.OBJECTIVE: To comparatively observe the injury of mitochondrial ultrastructure and the expressions of fas and bax in hippocampal tissue of rats with status epilepticus of different latency.DESIGN: A randomized control study.SETTING: Department of Anesthesiology and Department of Neurology, Qilu Hospital of Shandong University.MATERIALS: Totally 110 male adult SD rats of 260-300 g were used. Kainic acid was purchased from American Sigma Company.METHODS: The experiment was carried out in the Pathological Laboratory of Shandong Academy of Medical Sciences between March and July 2005.① Totally 100 SD rats were divided into two groups according to the random number table method:intraperitoneal injection group and caudal venous injection group.The rats were given kainic acid injected intraperitoneally(12 mg/kg)and through caudal vein (10 mg/kg) respectively. Each group was observed at 3, 6, 24, 48 and 72 hours after status epilepticus respectively.Ten rats were selected for each time point, including 2 for examination of electron microscope and 8 for the diction of the fas and bax mRNA expressions. The time and manifestations of seizure were observed, and the seizure was lasted for 2 hours, and then it was terminated by intraperitoneal injection of diazepam (10 mg/kg). Another 10 rats were used as the normal control group, and the materials were taken at 24 hours after status epilepticus, 2 of rats for the examination of electron microscope and 8 of them for the reverse transcription-polymerase chain reaction (RT-PCR). ② The ultrastructure of hippocampal neurons and its mitochondria were observed with transmission electron microscope. ③ The fas and bax mRNA expressions were detected with reverse transcription-polymerase chain reaction (RT-PCR).MAIN OUTCOME MEASURES: The

  19. Expresssion of bax/bcl-xl by low-power laser irradiation in the Amyloid Beta 25-35 induced apoptosis of PC12 cell

    Zhang, Lan; Xing, Da

    2006-09-01

    Apoptosis has been reported as a contributing pathophysiological mechanism of Alzheimer's disease (AD). Recently, the anti-apoptosis function of low-power laser irradiation (LPLI) was proposed, suggesting LPLI may become a new means for AD therapy. In this study, we aimed to demonstrate the anti-apoptosis function of LPLI at molecular level. Aβ 25-35 was used to induce apoptosis of PC12 cell, and then the cells were dealt with LPLI. After irradiation, the molecular level of apoptosis was detected by quantifying the bax I bcl-xl mRNA ratio using a highly sensitive and quantitative polymerase chain reaction (QT-PCR) technique. The primary results show that the bax Ibcl-xl mRNA ratio of the PC12 cell treated with Aβ 25-35 was decreased by LPLI, demonstrating the anti-apoptosis function of LPLI at molecular level.

  20. Titanium Dioxide (TiO2) Nanoparticles Preferentially Induce Cell Death in Transformed Cells in a Bak/Bax-Independent Fashion

    Yanglong Zhu; Eaton, John W.; Chi Li

    2012-01-01

    While the cytotoxic effects of titanium dioxide (TiO(2)) nanoparticles have been under intense investigation, the molecular mechanisms of this cytotoxicity remain unknown. Here we investigated the influence of oncogenic transformation and a major apoptotic signaling pathway on cellular responses to TiO(2) nanoparticles. Isogenic wild-type (WT) and apoptosis-resistant (Bak(-/-)Bax(-/-)) cell lines with and without tumorigenic transformation were examined. TiO(2) nanoparticles preferentially re...

  1. Adenosine induces cell cycle arrest and apoptosis via cyclinD1/Cdk4 and Bcl-2/Bax pathways in human ovarian cancer cell line OVCAR-3.

    Shirali, Saeid; Aghaei, Mahmoud; Shabani, Mahdi; Fathi, Mojtaba; Sohrabi, Majid; Moeinifard, Marzieh

    2013-04-01

    Adenosine is a regulatory molecule with widespread physiological effects in almost every cells and acts as a potent regulator of cell growth. Adenosine has been shown to inhibit cell growth and induce apoptosis in the several cancer cells via caspase activation and Bcl-2/Bax pathway. The present study was designed to understand the mechanism underlying adenosine-induced apoptosis in the OVCAR-3 human ovarian cancer cells. MTT viability, BrdU and cell counting assays were used to study the cell proliferation effect of adenosine in presence of adenosine deaminase inhibitor and the nucleoside transporter inhibitor. Cell cycle analysis, propidium iodide and annexin V staining, caspase-3 activity assay, cyclinD1, Cdk4, Bcl-2 and Bax protein expressions were assessed to detect apoptosis. Adenosine significantly inhibited cell proliferation in a concentration-dependent manner in OVCAR-3 cell line. Adenosine induced cell cycle arrest in G0/G1 phase via Cdk4/cyclinD1-mediated pathway. Adenosine induced apoptosis, which was determined by Annexin V-FITC staining and increased sub-G1 population. Moreover, down-regulation of Bcl-2 protein expression, up-regulation of Bax protein expression and activation of caspase-3 were observed in response to adenosine treatment. The results of this study suggest that extracellular adenosine induced G1 cell cycle arrest and apoptosis in ovarian cancer cells via cyclinD1/ Cdk4 and Bcl-2/Bax pathways and caspase-3 activation. These data might suggest that adenosine could be used as an agent for the treatment of ovarian cancer. PMID:23345014

  2. Disturbance of Bcl-2, Bax, Caspase-3, Ki-67 and C-myc expression in acute and subchronic exposure to benzo(a)pyrene in cervix.

    Gao, Meili; Li, Yongfei; Ji, Xiaoying; Xue, Xiaochang; Chen, Lan; Feng, Guodong; Zhang, Huqin; Wang, Huichun; Shah, Walayat; Hou, Zhanwu; Kong, Yu

    2016-03-01

    Epidemiological studies have demonstrated that cigarette smoking is an important cofactor or an independent risk factor for the development of cervical cancer. Benzo(a)pyrene (BaP) is one of the most potent tobacco smoke carcinogens in tobacco smoke. BaP induced DNA damage and over expression in p53 cervical tissue of mice as demonstrated in our previous study. Here we present the findings of exposure to BaP on the expression of Bcl-2, C-myc, Ki-67, Caspase-3 and Bax genes in mouse cervix. Acute intraperitoneal administration of BaP (12.5, 25, 50, 100mg/kg body weight) to ICR female mice induced a significant increase in Bcl-2, C-myc, Ki-67 mRNA and protein level till 72h except in Bcl-2 at 24h with 12.5, 25, 50mg/kg as well as at 48h with 12.5mg/kg body weight post treatment. A significant increase was also seen in Caspase-3 and Bax mRNA and protein level with peak level at 24h and gradual decrease till 72h, however, the expression of caspase-3 increased while that of Bax decreased with increasing dose of Bap after 24h. In sub chronic intraperitoneal and oral gavage administration of BaP (2.5, 5, 10mg/kg body weight), similar significant increase was observed for all the examined genes as compared to the control and vehicle groups, however the expression of Bax decreased in a dose dependent manner. The findings of this study will help in further understanding the molecular mechanism of BaP induced carcinogenesis of cervical cancer. PMID:26709117

  3. An Assay of Bax and Bcl2 Expression in Mice Hippocampus Following Ischemia-Reperfusion Treatment with CoQ10

    Jalal Hassanshahi

    2013-10-01

    Full Text Available Introduction: Preliminary studies confirmed reduction of cell death following treatment with antioxidants. According to this finding we investigated the relationship between consumption of CoQ10 and expression of bax and bcl2 in hippocampus ischemia that this expression related to cell programmed death.Material and Methods: We studied the protective role of CoQ10 against ischemia-reperfusion. Experimental design includes four groups: intact (N=7, ischemic control (N=7, sham control (N=7 and treatment groups with CoQ10 (N=7. The mice (treatment group treated with CoQ10 as Pre-Treatment for a week. Then, ischemia induced by common carotid artery ligation and following the reduction in inflammation (a week the treatment group post-treated with CoQ10 for a week. Nissl staining applied to counting necrotic cells of hippocampus and the western blotting performed to measurement the bax and bcl2 expression. Tunnel kit was used to quantify apoptotic cell death while to short term memory scale, we apply Y-maze.Results: Cell death was significantly lower when mice treated with CoQ10. Bax expression was significantly high in ischemic group but in treatment group was less and reversely the bcl2 expression in ischemic group was lower than treatment and vehicle groups. The memory test results were consistent with cell death results. Conclusion: Ischemia for 15 minutes induced cell death in hippocampus with more potent effect on CA1. CoQ10 intake significantly reduced cell death and decreased memory loss. with prevent of expression of bax and increase in expression of bcl2.

  4. Study of regulatory promoter polymorphism (-248 G>A) of Bax gene in patients with gastric cancer in the northern provinces of Iran

    Mirmajidi, Seyedeh Habibeh; Najafi, Mojtaba; Mirmajidi, Seyedeh Tahereh; Nasri Nasrabadi, Nafiseh

    2016-01-01

    Aim: The aim of this study is to evaluate the polymorphism in Bax gene and its association with some clinical pathology traits in gastric cancer. Background: Gastric cancer is considered as the fourth most common cancer in the north and northwest of Iran. Bcl2 family has a key role in regulation of apoptosis, and any changes in the expression of Bcl2 lead to cancer. Patients and methods: Blood samples were collected from 100 cases and 89 controls in the northern provinces of Iran to evaluate promoter polymorphism (-248GBax gene. Genotyping was carried out by PCR-RFLP method. Results: The result of this study demonstrated the existence of polymorphism in the above-mentioned region of Bax gene. Sixty-nine patients (%69) with genotype GG and 31 patients (%31) with genotype AG were observed in the case group. No mutant genotype was found among cases. Sixty-seven individuals (%75/28) with genotype GG, 21 individuals (%23/59) with genotype AG and only one mutant genotype (AA) were demonstrated in the control group. The bioinformatics analysis showed that this polymorphism removed the probable Sp1 motifs, which may affect its expression in the cells. Conclusion: Allele G was the most frequent between both patient and control samples. Polymorphism may be effective on Bax expression, but it requires further investigation. Our results showed significant effects between genotypes and features of gender and age, whereas no significant relation were observed between the genotypes and grade, stage as well as smoking traits. PMID:26744613

  5. Effect of Achyranthes Bidentata Polysaccharides on the Expression of BCL-2 and Bax in Hepatic Tissues after Exhaustive Exercise in Rats

    Lin, Jinyang; Zhang, Zhuoying; Shan, Ying

    2010-01-01

    This study aims to assess the effects of Achyranthes bidentata polysaccharides (ABPS) on the expression of bcl-2 and bax in hepatic tissues after exhaustive exercise in order to provide theoretical support for the application of ABPS in the field of sports nutrition. Thirty male Sprague-Dawley rats were randomized into three groups, each consisting of 10 rats: Normal control group (NCG), Exhausting exercises control group (EECG), ABPS treated group (ATG). ABPS were fed orally by gastric intub...

  6. Expression of bax and bcl2 Genes in MDMA-induced Hepatotoxicity on Rat Liver Using Quantitative Real-Time PCR Method through Triggering Programmed Cell Death

    2015-01-01

    Background: 3-4methylenedioxymethamphetamine (MDMA) is a synthetic and psychoactive drug, which is known popularly as Ecstasy and has toxic effects on human organs. Objectives: Considering the potential toxic interaction, this study was performed to quantify the expression of bax and bcl2 genes in MDMA-induced hepatotoxicity on rat liver. Subsequently, we evaluated pentoxifylline as a possible protective drug on hepatotoxicity. Materials and Methods: Adult male Wistar rats weighting 250 - 300...

  7. Oridonin induces apoptosis of HeLa cells via altering expres sion of Bcl-2/Bax and activating caspase-3/ICAD pathway

    Chun-ling ZHANG; Li-jun WU; Shin-ichi TASHIRO; Satoshi ONODERA; Takashi IKEJIMA

    2004-01-01

    AIM: To study the mechanisms by which oridonin inhibited HeLa cell growth in vitro. METHODS: Viability of oridonin-induced HeLa cells was measured by MTT assay. Apoptotic cells with condensed nuclei were visualized by phase contrast microscopy. Nucleosomal DNA fragmentation was assayed by agarose gel electrophoresis.Caspase activity was assayed using fiuorometric protease assay. ICAD, Bcl-2, and Bax proteins expression were detected by Western blot analysis. RESULTS: Oridonin induced oligonucleosomal fragmentation of DNA and increased caspase-3 activity, on the other hand, reduced the expression of inhibitor of caspase-3-activated DNase (ICAD), a caspase-3 substrate, at 12 h in HeLa cells. Oridonin-induced DNA fragmentation, caspase-3 activation and down-regulation of ICAD expression were effectively inhibited by a caspase-3 inhibitor, z-DEVD-fmk (z-AspGlu-Val-Asp-fmk). However, pretreatment with an inhibitor of poly (ADP-ribose) polymerase (PARP), 3, 4-dihydro5-[4-(1-piperidinyl)butoxy]-1 (2H)-isoquinolinone (DPQ), did not suppress oridonin-induced HeLa cell death. In addition, oridonin-induced apoptosis was associated with an increase in the expression of the apoptosis inducer Bax, and a significant reduction in expression of the apoptosis suppressor Bcl-2 in mitochondria. CONCLUSION:Oridonin induces HeLa cells apoptosis by altering balance of Bcl-2 and Bax protein expression and activation of caspase-3/ICAD pathway.

  8. Influence of 103Pd radioactive stent on the expression of Bcl-2 and Bax mRNA in vascular smooth muscle cells

    Objective: Observing the influence of 103Pd radioactive stent on the expression of apoptosis relative genes of Bcl-2 and Bax mRNA in rabbit from tunica media of abdominal aorta arteries by 103Pd radioactive stent after angioplasty. Methods: Fifty male New Zealand rabbits were randomized into an ordinary stent (OS) group, 103Pd stent (PdS) group and a control group. The aortas were harvested on days 3, 7, 14, 28 and 56 after operation. The pathomorphology observation was performed and the method of in situ hybridization was used to evaluate the expression of Bcl-2 and Bax mRNA. Results: The severity of the stenosis in the PdS group was less severe than that of the OS group. It was most obvious on the 56th day (P0.05). The ratio of the PdS group was much lower than that of the OS group on days 3 to 28 (P103Pd radioactive stent increases the expression of Bax mRNA which stimulates apoptosis and decreases the expression of Bcl-2 mRNA which inhibits apoptosis, resulting in improvement of the ratio of apoptosis to proliferation and prevention of the occurrence of restenosis. (author)

  9. Gambogic acid induces mitochondria-dependent apoptosis by modulation of Bcl-2 and Bax in mantle cell lymphoma JeKo-1 cells

    Jingyan Xu; Min Zhou; Jian Ouyang; Jing Wang; Qiguo Zhang; Yong Xu; Yueyi Xu

    2013-01-01

    Objective:To study the mechanisms in gambogic acid (GA)-induced JeKo-1 human Mantle Cell Lymphoma cell apoptosis in vitro.Methods:The proliferation of GA-treated JeKo-1 cells was measured by CCK-8 assay and Ki-67 immunocytochemical detection.Apoptosis,cell cycle and mitochondrial membrane potential were measured by flow cytometric analysis.Caspase-3,-8 and-9 were detected by colorimetric assay.Bcl-2 and Bax were analyzed by Western blotting.Results:GA inhibited cell growth in a time-and dose-dependent manner.GA induces apoptosis in JeKo-1 cells but not in normal bone marrow cells,which was involved in reducing the membrane potential of mitochondria,activating caspases-3,-8 and-9 and decreasing the ratio of Bcl-2 and Bax without cell cycle arresting.Conclusions:GA induced apoptosis in human MCL JeKo-1 cells by regulating Bcl-2/Bax and activating caspase-3,-8 and-9 via mitochondrial pathway without affecting cell cycle.

  10. Effect of Hypoxic Preconditioning on Neural Cell Apoptosis and Expression of Bcl-2 and Bax in Cerebral Ischemia-Reperfusion in Rats

    2006-01-01

    In order to investigate the protective effect of hypoxic preconditioning on the cerebral ischemia-reperfusion injury, the expression of Bcl-2 and Bax was detected by using immunohistochemical staining after 3 h cerebral ischemia followed by 1, 6, 12, 24 and 48 h reperfusion respectively in rats treated with or without hypoxic preconditioning before cerebral ischemia. In addition,the apoptosis of neural cells and the behavioral scores for neurological functions recovery were evaluated by TUNEL staining and "crawvling method", respectively. Compared with control group (cerebral ischemia-reperfusion without hypoxic preconditioning), the expression of Bcl-2 was significantly increased, but that of Bax decreased in the hypoxic preconditioning group (cerebral ischemiareperfusion with hypoxic preconditioning), both P<0. 05. The pre-treatment with hypoxic preconditioning could reduce the apoptosis of neural cells and promote the neurological function recovery as compared to control group. It was suggested that hypoxic preconditioning may have protective effects on the cerebral ischemia-reperfusion injury by inhibiting the apoptosis of neural cells, increase the expression of Bcl-2 and decrease the expression of Bax.

  11. Curcumin induces the expression of NF-κB and Bcl-2/Bax in human renal cell carcinoma cell line ACHN

    Gang Li; Tie Chong; Ziming Wang

    2009-01-01

    Objective: To explore the in vitro effects of curcumin on the proliferation and apoptosis of the human renal cell carcinoma cell line ACHN, and to investigate its mechanisms of action. Methods: The human renal cell carcinoma cell line ACHN was treated with different concentrations of curcumin for 24 h. The MTT assay was used to evaluate the cytotoxic effects of curcumin and flow cytometry was utilized to observe and detect the apoptosis of ACHN cells induced by curcumin. The expression levels of Bcl-2, Bax and NF-κBP65 mRNA were evaluated by Reverse Transcription-Polymerase Chain Reaction (RT-PCR), while the expression of Bcl-2, Bax, NF-κBP65 and IkB proteins was evaluated by Western blot. Results: The concentrations of curcumin used significantly inhibited the proliferation of ACHN human renal cell carcinoma cells in vitro in a dose and time-dependent manner (Ftime=5.55, P < 0.05; Fdose=110.05, P < 0.05). Obvious apoptosis of cells treated with different concentrations of curcumin could be observed by FCM. Compared with the control group, the apoptosis rates of curcumin-treated cells were markedly increased (F=96.35, P < 0.05). Lower dose of curcumin significantly induced the apoptosis of ACHN cells. With intervention of different concentrations of curcumin (0, 10, 20 and 40 μmol/L) for 24 h, the expression levels of Bcl-2 and NF-κBP65 mRNA in ACHN cells were decreased while the expression level of Bax mRNA was increased (P < 0.05), and Bcl-2, and NF-κBP65 protein decreased, while Bax and IκB protein increased compared with those in the untreated group. Conclusion: Curcumin inhibited proliferation and increased apoptosis of the human renal cell carcinoma cell line ACHN. These curcumin effects appear to involve up-regulating IκB, down-regulating NF-κB, and regulating the expression of the apoptosis genes Bcl-2/Bax.

  12. Expression of anti-apoptotic protein Bcl-2 and pro-apoptotic protein Bax after delayed paraplegia induced by ischemia/reperfusion injury

    Bibo Liu; Miao Liu; Duoning Wang; Wei Ma; Shengli Dang

    2006-01-01

    BACKGROUND:Operation of spine does not involve in spinal cord;however,spinal cord injury occurs at post-operation induced by unclear factors.Meanwhile,after decompression of lumbar spinal canal,symptoms are severer and severer.In addition,during extirpation of oervical and lumbar intervertabral disc,spinal cord and its vessels are not damaged,but spinal cord injury is also suffered from patients with partial improvement.All statuses mentioned above are related to expressed changes of bc/-2 gene inhibiting apoptosis and bax gene accelerating apoptosis.OBJECTIVE:To observe motor function of hindlimbs of ischemia/reperfusion model in rabbits at various time points after reperfusion and expressions of apoptosis correlated protein Bcl-2 and Bax.DESIGN:Completely randomized grouping design and contrast study.SEITING:Department of Orthopedics,the First Affiliated Hospital of Medical School,Xi'an Jiao Tong University.MATERIALS:Forty-eight New Zealand white rabbits of both genders were randomly divided into sham operation group(n=24) and model group(n=24),and then,rabbits in each group were observed at four time points:8,24,72 and 168 hours after reperfusion,with 6 in each time point.Rabbit-anti-rabbit Bcl-2 antibody and rabbit-anti-rabbit Bax antibody were provided by Boster Company.The procedures were accordant to the METHODS:The experiment was carried out at Laboratory of Orthopaedics of First Affiliated Hospital of Medical School,Xi'an Jiao Tong University from April to August 2005.①Delayed paralysis models of spinal cord after ischemia/reperfusion were established based on method of Zivin et al.Animals in sham operation group underwent an exposure of abdominal aorta but the aorta was not occluded.②Motor function of hindlimb was observed 8,24,72 and 168 hours after reperfusion.A grade of 0-5 was assigned to each animal (grade 0:no voluntary hind limb function;grade 5:normal hop;grades 0-3:paraplegia).③The lumbar segment of the spinal cord(L3 to L5)was used for

  13. Exercise Preconditioning Decreases the Ratio of bax/bcl-2 mRNA Expression in Midbrain of Mouse Model with Parkinson's Disease%运动预适应下调帕金森小鼠中脑bax/bcl-2 mRNA表达比值

    薛宏斌; 张勇; 刘洪涛; 马强

    2009-01-01

    目的:探讨运动预适应是否下调帕金森小鼠中脑细胞bax/bcl-2 mRNA表达比值.方法:32只雄性小鼠(8~9周龄)首先随机分为安静组、运动组.运动组连续5周,每天1次中等强度(12米/分,20分钟)跑台训练.然后以上两组再各自随机分成两组,于训练结束后第2天分别注射生理盐水或中等剂量MPTP(30mg/kg×2次).只有表现典型行为变化的MPTP小鼠被认为是成功帕金森模型小鼠,进入最终分组,分为安静+生理盐水组(Se+Sa)、运动+生理盐水组(E+Sa)、安 +MPTP组(Se+M)、运 +MPTP组(E+M)4组,每组8只.训练结束后第11天处死动物,检测小鼠中脑bax、bcl-2 mRNA表达,计算bax/bcl-2 mRNA表达比值.结果:E+M组小鼠中脑bax mRNA表达较Se+M组显著降低(P<0.05),E+M组bcl-2 mRNA表达较Se+M组显著增加(P<0.05),E+M组bax/bcl-2比值显著小于Se+M组(P<0.05).结论:运动预适应改变帕金森小鼠中脑黑质细胞凋亡信号分子bcl-2、bax mRNA表达,降低bax/bcl-2 mRNA表达比值.

  14. Ginkgo biloba leaf extract effects on inducible nitric oxide synthase, Bcl-2, and Bax expression in rat models of spinal cord injury

    Jiejun Jiao; Bin Du

    2008-01-01

    BACKGROUND: Ginkgo biloba leaf extract exhibits neuroprotective effects in spinal cord injury. However,the mechanisms of action remain unclear.OBJECTIVE: To investigate inducible nitric oxide synthase (iNOS) and Bcl-2/Bax expression in the injured spinal cord, and to explore the neuroprotective mechanisms of ginkgo biloba leaf extract in rats with spinal cord injury.DESIGN, TIME AND SETTING: The randomized, controlled, cell molecular biology experiment was performed at Soochow University, China from March 2007 to March 2008.MATERIALS: A total of 120 healthy, adult Sprague Dawley rats were selected for this study. Rat models of moderate acute thoracic (T9) spinal cord injury were established using the modified Allen method.Shuxuening injection was obtained from Zhenbaodao Pharmaceutical Co., Ltd., China. Methylprednisolone was purchased from North China Pharmaceutical Co., Ltd.METHODS: All rats were equally and randomly divided into four groups. Only the spinal cord was exposed in the sham operation group rats. In the trauma group, rats were not treated with drugs following spinal cord injury. Rats in the hormone group were intraperitoneally injected with 30 mg/kg methylprcdnisolone following spinal cord injury. Rats in the ginkgo biloba leaf extract group were intraperitoneally infused with a 1.0 mL/kg Shuxuening injection per day.MAIN OUTCOME MEASURES: At l hour, as well as 1, 3, 5, 7, and 14 days after spinal cord injury,iNOS- and Bcl-2/Bax-positive cells were quantified with immunohistochemistry. Pathological changes were detected using hematoxylin-eosin staining under an optical microscope.RESULTS: Spinal cord injury in the ginkgo biloba leaf extract and hormone groups was milder compared with the trauma group. Demyelination was significantly ameliorated and the necrotic cavity was obviously reduced in the injured spinal cord of rats in the ginkgo biloba leaf extract and hormone groups at each time point, iNOS expression was increased in the injured spinal cord

  15. Multiple doses of erythropoietin impair liver regeneration by increasing TNF-alpha, the Bax to Bcl-xL ratio and apoptotic cell death.

    Katja Klemm

    Full Text Available BACKGROUND: Liver resection and the use of small-for-size grafts are restricted by the necessity to provide a sufficient amount of functional liver mass. Only few promising strategies to maximize liver regeneration are available. Apart from its erythropoiesis-stimulating effect, erythropoietin (EPO has meanwhile been recognized as mitogenic, tissue-protective, and anti-apoptotic pleiotropic cytokine. Thus, EPO may support regeneration of hepatic tissue. METHODOLOGY: Rats undergoing 68% hepatectomy received daily either high dose (5000 IU/kg bw i.v. or low dose (500 IU/kg bw i.v. recombinant human EPO or equal amounts of physiologic saline. Parameters of liver regeneration and hepatocellular apoptosis were assessed at 24 h, 48 h and 5 d after resection. In addition, red blood cell count, hematocrit and serum EPO levels as well as plasma concentrations of TNF-alpha and IL-6 were evaluated. Further, hepatic Bcl-x(L and Bax protein expression were analyzed by Western blot. PRINCIPAL FINDINGS: Administration of EPO significantly reduced the expression of PCNA at 24 h followed by a significant decrease in restitution of liver mass at day 5 after partial hepatectomy. EPO increased TNF-alpha levels and shifted the Bcl-x(L to Bax ratio towards the pro-apoptotic Bax resulting in significantly increased hepatocellular apoptosis. CONCLUSIONS: Multiple doses of EPO after partial hepatectomy increase hepatocellular apoptosis and impair liver regeneration in rats. Thus, careful consideration should be made in pre- and post-operative recombinant human EPO administration in the setting of liver resection and transplantation.

  16. Punicalagin attenuated cerebral ischemia-reperfusion insult via inhibition of proinflammatory cytokines, up-regulation of Bcl-2, down-regulation of Bax, and caspase-3.

    Yaidikar, Lavanya; Thakur, Santhrani

    2015-04-01

    Punicalagin (PG) is a hydrolysable tannin compound found in Punica granatum L. The purpose of the present work is to explore the neuroprotective mechanism of PG against ischemia-reperfusion (I/R) injury in rat model of middle cerebral artery occlusion (MCAO). Rats were randomly divided into sham, MCAO, and PG-treated groups. PG (15 and 30 mg/kg), the vehicle was administered orally for 7 days prior to MCAO. Rats were anesthetised with ketamine (100 mg/kg/im), xylazine (10 mg/kg/im) and subjected to 2 h occlusion and 22 h reperfusion. The effects of PG on behavioral deficit and infarct volume, the levels of glutamate and calcium as well as the levels of inflammatory cytokines tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6) were evaluated. Moreover, the expressions of caspase-3, Bcl-2, and Bax were detected by Western blotting. As compared with MCAO group, PG-treated rats showed dose-dependent reduction in infarct volume and substantial improvement in behavioral deficit. The levels of glutamate, calcium, TNF-α, IL-1β, and IL-6 were restored significantly. The Western blotting results revealed that the expression of Bcl-2 was up-regulated and that of caspase-3, Bax were down-regulated when exposed to PG. From our results, it can be concluded that PG showed an ameliorative effect against cerebral I/R injury in rats through its anti-inflammatory, antioxidant actions besides it inhibits excitotoxicity. It also suppresses apoptosis through regulating, Bcl-2, caspase-3, and Bax protein expressions, perhaps another mechanism by which PG employs its neuroprotective action. PMID:25555468

  17. Garlic ((Allium sativum)) Fresh Juice Induces Apoptosis in Human Oral Squamous Cell Carcinoma: The Involvement of Caspase-3, Bax and Bcl-2.

    Farhadi, Farrokh; Jahanpour, Salar; Hazem, Kameliya; Aghbali, Amirala; Baradran, Behzad; Vahid Pakdel, Seyyed Mahdi

    2015-01-01

    Background and aims. There is no report on the apoptotic impact of Allium sativum L.(Garlic) on the oral squamous cell carcinoma (KB); hence, this study was designed to survey the apoptotic effects of garlic fresh juice (GFJ) on the KB cells. Materials and methods. MTTassay (MicrocultureTetrazolium Assay) was carried out to evaluate the cytotoxicity of GFJ on KB cells. Furthermore, TUNEL(Terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling)and DNA fragmentation tests were performed to determine if GFJ is able to induce apoptosis in KB cells. Also a standard kit was used to assess caspase-3 activity in KB cells. Also western blotting was employed to evaluate the effect of GFJ on Bax:Bcl-2 ratio. Results. Significant cytotoxic effects were observed for the minimum used concentration (1μg/mL) as calculated to be 77.97±2.3% for 24 h and 818±3.1% for 36h of incubation (P < 0.001). Furthermore, TUNEL and DNA fragmentation tests corroborated the apoptosis inducing activity of GFJ. Consistently, after treating KB cells with GFJ(1μg/mL), caspase-3 activity and Bax:Bcl-2 ratio were raised by 7.3±0.6 and (P <0.001) folds, respectively. Conclusion. The results of this study advanced that GFJ induces apoptosis in the KB cells through increasing caspase-3 activity and Bax:Bcl2 ratio which could be attributed to its organo-sulfurcomponents. PMID:26889365

  18. Dexamethasone protected human glioblastoma U87MG cells from temozolomide induced apoptosis by maintaining Bax:Bcl-2 ratio and preventing proteolytic activities

    Patel Sunil J

    2004-12-01

    Full Text Available Abstract Background Glioblastoma is the deadliest and most prevalent brain tumor. Dexamethasone (DXM is a commonly used steroid for treating glioblastoma patients for alleviation of vasogenic edema and pain prior to treatment with chemotherapeutic drugs. Temozolomide (TMZ, an alkylating agent, has recently been introduced in clinical trials for treating glioblastoma. Here, we evaluated the modulatory effect of DXM on TMZ induced apoptosis in human glioblastoma U87MG cells. Results Freshly grown cells were treated with different doses of DXM or TMZ for 6 h followed by incubation in a drug-free medium for 48 h. Wright staining and ApopTag assay showed no apoptosis in cells treated with 40 μM DXM but considerable amounts of apoptosis in cells treated with 100 μM TMZ. Apoptosis in TMZ treated cells was associated with an increase in intracellular free [Ca2+], as determined by fura-2 assay. Western blot analyses showed alternations in the levels of Bax (pro-apoptotic and Bcl-2 (anti-apoptotic proteins resulting in increased Bax:Bcl-2 ratio in TMZ treated cells. Western blot analyses also detected overexpression of calpain and caspase-3, which cleaved 270 kD α-spectrin at specific sites for generation of 145 and 120 kD spectrin break down products (SBDPs, respectively. However, 1-h pretreatment of cells with 40 μM DXM dramatically decreased TMZ induced apoptosis, decreasing Bax:Bcl-2 ratio and SBDPs. Conclusion Our results revealed an antagonistic effect of DXM on TMZ induced apoptosis in human glioblastoma U87MG cells, implying that treatment of glioblastoma patients with DXM prior to chemotherapy with TMZ might result in an undesirable clinical outcome.

  19. Suppression of the Arboviruses Dengue and Chikungunya Using a Dual-Acting Group-I Intron Coupled with Conditional Expression of the Bax C-Terminal Domain.

    James R Carter

    Full Text Available In portions of South Asia, vectors and patients co-infected with dengue (DENV and chikungunya (CHIKV are on the rise, with the potential for this occurrence in other regions of the world, for example the United States. Therefore, we engineered an antiviral approach that suppresses the replication of both arboviruses in mosquito cells using a single antiviral group I intron. We devised unique configurations of internal, external, and guide sequences that permit homologous recognition and splicing with conserved target sequences in the genomes of both viruses using a single trans-splicing Group I intron, and examined their effectiveness to suppress infections of DENV and CHIKV in mosquito cells when coupled with a proapoptotic 3' exon, ΔN Bax. RT-PCR demonstrated the utility of these introns in trans-splicing the ΔN Bax sequence downstream of either the DENV or CHIKV target site in transformed Aedes albopictus C6/36 cells, independent of the order in which the virus specific targeting sequences were inserted into the construct. This trans-splicing reaction forms DENV or CHIKV ΔN Bax RNA fusions that led to apoptotic cell death as evidenced by annexin V staining, caspase, and DNA fragmentation assays. TCID50-IFA analyses demonstrate effective suppression of DENV and CHIKV infections by our anti-arbovirus group I intron approach. This represents the first report of a dual-acting Group I intron, and demonstrates that we can target DENV and CHIKV RNAs in a sequence specific manner with a single, uniquely configured CHIKV/DENV dual targeting group I intron, leading to replication suppression of both arboviruses, and thus providing a promising single antiviral for the transgenic suppression of multiple arboviruses.

  20. Oxidized low-density lipoprotein induces apoptotic insults to mouse cerebral endothelial cells via a Bax-mitochondria-caspase protease pathway

    Cerebral endothelial cells (CECs) are crucial components of the blood-brain barrier. Oxidized low-density lipoprotein (oxLDL) can induce cell injuries. In this study, we attempted to evaluate the effects of oxLDL on mouse CECs and its possible mechanisms. Mouse CECs were isolated from brain tissues and identified by immunocytochemical staining of vimentin and Factor VIII. oxLDL was prepared from LDL oxidation by copper sulfate. Exposure of mouse CECs to oxLDL decreased cell viability in concentration- and time-dependent manners. oxLDL time-dependently caused shrinkage of cell morphologies. Administration of oxLDL to CECs induced DNA fragmentation in concentration- and time-dependent manners. Analysis of the cell cycle revealed that oxLDL concentration- and time-dependently increased the proportion of CECs which underwent apoptosis. Analysis of confocal microscopy and immunoblot revealed that oxLDL significantly increased cellular and mitochondrial Bax levels as well as the translocation of this proapoptotic protein from the cytoplasm to mitochondria. In parallel with the increase in the levels and translocation of Bax, oxLDL time-dependently decreased the mitochondrial membrane potential. Exposure of mouse CECs to oxLDL decreased the amounts of mitochondrial cytochrome c, but enhanced cytosolic cytochrome c levels. The amounts of intracellular reactive oxygen species were significantly augmented after oxLDL administration. Sequentially, oxLDL increased activities of caspase-9, -3, and -6 in time-dependent manners. Pretreatment with Z-VEID-FMK, an inhibitor of caspase-6, significantly decreased caspase-6 activity and the oxLDL-induced DNA fragmentation and cell apoptosis. This study showed that oxLDL induces apoptotic insults to CECs via signal-transducing events, including enhancing Bax translocation, mitochondrial dysfunction, cytochrome c release, increases in intracellular reactive oxygen species, and cascade activation of caspase-9, -3, and -6. Therefore, ox

  1. Multiple broken symmetries in striped La2 -xBaxCuO4 detected by the field-symmetric Nernst effect

    Soumyanarayanan, Anjan; Tee, X. Y.; Ito, T.; Ushiyama, T.; Tomioka, Y.; Panagopoulos, C.

    2016-02-01

    We report a thermoelectric investigation of the stripe and superconducting phases of the cuprate La2 -xBaxCuO4 near x =1 /8 . We vary the doping and the magnetic field to identify features in the field-symmetric Nernst effect consistent with the reports of time-reversal symmetry breaking above the superconducting Tc. Crucially, we further detect a field-invariant peak at the stripe charge order temperature, TCO. Our observations suggest the onset of a nontrivial charge ordered phase at TCO, and the subsequent presence of spontaneously generated vortices before the emergence of bulk superconductivity.

  2. Apoptosis and the activity of ceramide, Bax and Bcl-2 in the lungs of neonatal rats exposed to limited and prolonged hyperoxia

    Bitar Fadi F; Nasser Michel; Panjarian Shoghag; Khayat Aline; Bitar Hala; Dbaibo Ghassan S; Husari Ahmad W; El-Sabban Marwan; Zaatari Ghazi; Mroueh Salman M

    2006-01-01

    Abstract Background The aim of the study is to examine the effect of limited and prolonged hyperoxia on neonatal rat lung. This is done by examining the morphologic changes of apoptosis, the expression of ceramide, an important mediator of apoptosis, the expression of inflammatory mediators represented by IL-1β and the expression of 2 proto-oncogenes that appear to modulate apoptosis (Bax and Bcl-2). Methods Newborn rats were placed in chambers containing room air or oxygen above 90% for 7 da...

  3. Phase diagrams and dielectric response of epitaxial (BaxSr1-x)TiO3 ultrathin films: A first-principles study

    A method based on first-principles calculations was used to construct temperature versus misfit strain phase diagrams for epitaxial (BaxSr1-x)TiO3 ultrathin films over the entire composition range from x=0 to x=1. The predicated phase diagrams show “topologies” that are similar to those calculated by Shirokov et al. (Phy. Rev. B. 79 (2009) 144118), but with quantitative differences that are examined and explained. The dependence of the dielectric permittivity on the misfit strain and the film composition at room temperature are also investigated and compared with available theoretical predications and experimental measurements.

  4. Structural and energetical studies of the adsorption of para and meta-isomers of xylene on pre-hydrated zeolite BaX. Characterization by neutron diffraction and temperature programmed desorption; Etude structurale et energetique de l'adsorption des isomeres para- et meta- du xylene dans la zeolithe BaX prehydratee. Caracterisation par diffraction des neutrons et thermodesorption programmee

    Pichon, Ch.

    1999-10-19

    The separation of p-xylene from C{sub 8} aromatics is performed industrially by selective adsorption on zeolitic materials. FAU-type zeolites are currently used for this separation and especially the partially hydrated BaX. The aim of this work is to characterize from a structural (by low temperature neutron powder diffraction) and an energetical (by temperature programmed desorption) point of view, the adsorption of para- and meta- isomers of xylene, for different fillings, as pure substances as well as mixtures, on pre-hydrated zeolite BaX. The influence of the water pre-adsorption on xylene adsorption selectivity is carefully discussed. The crystalline structure of the zeolite BaX (framework and compensation of charge cations) and of the adsorbed phase (water, p- and m-xylene molecules) are completely characterized by neutron diffraction. The location and the distribution of water and xylene molecules on their adsorption sites is especially followed as a function of the filling of the zeolite and of the composition of the adsorbed phase. Microscopic measurements were correlated to the energetical analysis (at a macroscopic level) in order to obtain a consistent description of adsorption phenomenon and to propose a possible origin for adsorption selectivity.

  5. Levofloxacin increases the effect of serum deprivation on anoikis of rat nucleus pulposus cells via Bax/Bcl-2/caspase-3 pathway.

    Yang, Si-Dong; Bai, Zhi-Long; Zhang, Feng; Ma, Lei; Yang, Da-Long; Ding, Wen-Yuan

    2014-12-01

    Levofloxacin, a fluoroquinolone, is a widely-used and effective antibiotic. However, various adverse side effects are associated with levofloxacin. The purpose of this study was to further explore the effects of levofloxacin on rat nucleus pulposus cells (NPCs). Inverted phase-contrast microscopy, flow cytometry and caspase-3 activity assays were used and revealed that serum deprivation induced apoptosis, which was markedly increased by levofloxacin in a dose-dependent manner. Simultaneously, levofloxacin decreased cell binding to type II collagen (COL2). Thus, levofloxacin-induced apoptosis exhibits characteristics of anoikis, the process by which cell death is triggered by separation from the extracellular matrix, which contains COL2. Furthermore, real-time quantitative RT-PCR was used to further confirm that levofloxacin downregulates COL2 expression in a dose-dependent manner. At last, western blot was used to find that levofloxacin increased the ratio of Bax/Bcl-2 and active caspase-3 in a dose-dependent manner. Levofloxacin therefore increases the effects of serum deprivation on anoikis by downregulating COL2 in rat NPCs in vitro via Bax/Bcl-2/caspase-3 pathway. This research provides a novel insight into the mechanisms of levofloxacin-induced toxicity and may potentially lead to a better understanding of the clinical effects of levofloxacin, especially in terms of intervertebral disc degeneration. PMID:25224805

  6. Inotodiol inhabits proliferation and induces apoptosis through modulating expression of cyclinE, p27, bcl-2, and bax in human cervical cancer HeLa cells.

    Zhao, Li-Wei; Zhong, Xiu-Hong; Yang, Shu-Yan; Zhang, Yi-Zhong; Yang, Ning-Jiang

    2014-01-01

    Inonotus obliquus is a medicinal mushroom that has been used as an effective agent to treat various diseases such as diabetes, tuberculosis and cancer. Inotodiol, an included triterpenoid shows significant anti-tumor effect. However, the mechanisms have not been well documented. In this study, we aimed to explore the effect of inotodiol on proliferation and apoptosis in human cervical cancer HeLa cells and investigated the underlying molecular mechanisms. HeLa cells were treated with different concentrations of inotodiol. The MTT assay was used to evaluate cell proliferating ability, flow cytometry (FCM) was employed for cell cycle analysis and cell apoptosis, while expression of cyclinE, p27, bcl-2 and bax was detected by immunocytochemistry. Proliferation of HeLa cells was inhibited by inotodiolin a dose-dependent manner at 24h (r=0.9999, pInonotus obliquus inhibited the proliferation of HeLa cells and induced apoptosis in vitro. The mechanisms may be related to promoting apoptosis through increasing the expression of bax and cutting bcl-2 and affecting the cell cycle by down-regulation the expression of cyclin E and up-regulation of p27. The results further indicate the potential value of inotodiol for treatment of human cervical cancer. PMID:24815470

  7. Hsp105 family proteins suppress staurosporine-induced apoptosis by inhibiting the translocation of Bax to mitochondria in HeLa cells

    Hsp105 (Hsp105α and Hsp105β), major heat shock proteins in mammalian cells, belong to a subgroup of the HSP70 family, HSP105/110. Previously, we have shown that Hsp105α has completely different effects on stress-induced apoptosis depending on cell type. However, the molecular mechanisms by which Hsp105α regulates stress-induced apoptosis are not fully understood. Here, we established HeLa cells that overexpress either Hsp105α or Hsp105β by removing doxycycline and examined how Hsp105 modifies staurosporine (STS)-induced apoptosis in HeLa cells. Apoptotic features such as the externalization of phosphatidylserine on the plasma membrane and nuclear morphological changes were induced by the treatment with STS, and the STS-induced apoptosis was suppressed by overexpression of Hsp105α or Hsp105β. In addition, we found that overexpression of Hsp105α or Hsp105β suppressed the activation of caspase-3 and caspase-9 by preventing the release of cytochrome c from mitochondria. Furthermore, the translocation of Bax to mitochondria, which results in the release of cytochrome c from the mitochondria, was also suppressed by the overexpression of Hsp105α or Hsp105β. Thus, it is suggested that Hsp105 suppresses the stress-induced apoptosis at its initial step, the translocation of Bax to mitochondria in HeLa cells

  8. Knockdown of miR-221 promotes the cisplatin-inducing apoptosis by targeting the BIM-Bax/Bak axis in breast cancer.

    Ye, Zhiqiang; Hao, Rutian; Cai, Yefeng; Wang, Xiaobo; Huang, Guanli

    2016-04-01

    Accumulating evidence shows that microRNAs (miRNAs) have a critical role in the initiation and progression of types of human cancer, including breast cancer. Recent research indicated that miRNAs are also related with the chemotherapy on cancers. In this study, the expression of miR-221 in breast cancer (BC) patients' serum and cancer tissues was found to be significantly up-regulated. The results of in vitro MTT assay indicated that although the anti-miR-221 oligonucleotide alone did not influence the viability of BC cell lines markedly, it significantly promoted the cytotoxicity of cisplatin (DDP) to BC cells. Mechanistic studies demonstrated that the gene of BIM (Bcl-2 interacting mediator of cell death), a pro-apoptotic Bcl-2 family protein, was up-regulated by the knockdown of miR-221. We found that the synergetic effect of anti-miR-221 on increasing the sensitivity of MDA-MB-231 was BIM dependant. Furthermore, results of immunoprecipitation showed the up-regulated BIM directly combined with the Bax and Bak, leading to mitochondrial dysfunction. Our results suggest the anti-miR-221 could promote the cisplatin-inducing apoptosis by targeting the Bim-Bax/Bak axis in breast cancer. PMID:26503209

  9. A- and B-site doping effect on physicochemical properties of Sr2‑xBaxMMoO6 (M = Mg, Mn, Fe) double perovskites — candidate anode materials for SOFCs

    Zheng, Kun; Świerczek, Konrad

    2016-06-01

    In this work, we evaluate the physicochemical properties of Sr2‑xBaxMMoO6 (M = Mg, Mn, Fe) double perovskites as alternative anode materials for solid oxide fuel cells, for which the effect of substitution of strontium by barium in a full range of compositions is studied. The crystal structure, microstructure, characterization of transport properties (electrical conductivity, Seebeck coefficient) and oxygen content as a function of temperature, as well as chemical stability in oxidizing and reducing conditions are discussed. Fe- and Mo-containing Sr2‑xBaxFeMoO6 oxides show very high total conductivities with values of 100-1000 Sṡcm‑1, while Sr2‑xBaxMgMoO6 present good redox stability.

  10. Expression of protein encoded by apoptosis-associated gene p53, bcl-2, and bax in adaptive response of thymocyte apoptosis in mice induced by low dose radiation with X-rays

    Objective: To explore the regulative mechanism of apoptosis-associated gene proteins on the adaptive response of thymocyte apoptosis in mice induced by low dose radiation with X-rays. Methods: Kunming male mice were irradiated with the inductive doses (D1: 25, 50, 75, 100 and 200 mGy; dose rate: 12.5 mGy ·min-1) and the challenging dose (D2: 1.5 Gy; dose rate: 287 mGy·min-1). The time interval between D1 and D2 was 6 h. The expressive levels of thymocyte apoptosis-associated gene proteins were measured with flow cytometry. Results: As compared with the sham-irradiation, the positive percentage of thymocyte Bcl-2 protein expression decreased significantly in D2 group (P<0.05), Bax increased significantly (P<0.05), and Bcl-2/Bax decreased significantly (P<0.001); p 53 increased significantly (P<0.001). As compared with D2 group, the positive percentage of thymocyte Bcl-2 protein expression increased in varying degree in D1+ D2 group of 25-75 mGy D1, Bax decreased in varying degree, and Bcl-2/Bax increased significantly (P<0.01); p53 decreased significantly (P<0.001 or P<0.05). Conclusion: The apoptotic thymocytes in the adaptive response of thymocyte apoptosis in mice induced by irradiation with 25-75 mGy decrease significantly due to the increase of apoptosis-associated gene Bcl-2 protein expression and Bcl-2/Bax, the decrease of Bax and p53 protein expressions. (authors)

  11. Apoptosis, mitosis, bcl-2, bax, and bcl-x as predictors of tumor response in stage I and II follicular lymphoma

    Purpose: Levels of spontaneous apoptosis predict for tumor responsiveness to radiation and chemotherapy in various animal systems. Oncogenes belonging to the bcl-2 family have been found to govern apoptosis, and bcl-2 has been shown to convey multi-drug resistance in lymphoma in vitro. To investigate the potential role of apoptosis and oncogenes involved in apoptosis as a predictors of response in human tumors, a retrospective review was undertaken of patients with Stage I and II follicular lymphoma in whom long term results were recently reported. Materials and Methods: The H and E slides of the initial specimens of 91 patients were obtained. All slides were reviewed by one pathologist (WCP) and representative sections were chosen. Twenty-four patients were treated with regional radiotherapy, 57 patients with multiagent chemotherapy and regional radiotherapy, and 10 patients with multiagent chemotherapy alone. Apoptotic index (AI) and mitotic index (MI) were determined for each tumor by counting the number of apoptotic bodies and mitotic cells present in 500 cells (100 cells in each of 5 follicles). Paraffin-embedded tissue was obtained for 52 of the initial specimens, and bcl-2, bax, and bcl-x status was determined by immunohistochemistry for these cases. The above parameters were correlated with overall survival (OS) and freedom from progression (FFP). Follow-up for living patients ranged from 51 to 212 months with a median value of 114 months. Results: The AI and MI values were low, ranging from 0 - 5.2% and 0 - 1.0%, respectively. The median AI value was 0.4%, and 63 patients had a MI of 0%. Patients who were treated with combined chemotherapy and radiotherapy had a higher FFP when their tumors had spontaneous levels of apoptosis of 0 had an improved FFP (83% vs 27% 10-year FFP; p=0.05). Bcl-2, bax and bcl-x staining was positive for 41, 48, and 47 initial specimens, respectively. No correlation of bcl-2, bax or bcl-x staining with clinical outcome was found

  12. Suppression of p53 and Bax accumulation after X-irradiation by small dose preirradiation in radioadaptive survival response of C57BL/6 mice

    It has been reported by the authors that X-irradiation with 0.30-0.50 Gy two weeks before a second exposure to a midlethal dose significantly reduced the bone marrow death rate in ICR and C57BL/6 strains of mice. Though radioadaptive response within blood-forming tissues has been speculated on the induction the radioresistance, molecular mechanism has been unknown yet. The present study was carried out to elucidate molecular mechanisms for the radioadaptive survival response. Effect of preirradiation (0.45 Gy) on p53 as well as Bax accumulation after challenging irradiation was studied in C57BL/6N mice. Accumulation of p53 was examined by the Western blotting method in the brain, thymus, lung, liver, intestine, spleen and bone-marrow of mice collected 5 hrs after challenging irradiation with 3 Gy. The challenging irradiation increased p53. The priming irradiation itself did not show any change in p53 level 2 weeks later. p53 accumulation after challenging irradiation with 3 Gy was significantly suppressed by the preirradiation in the spleen, among the tissues examined. Suppression of p53 by preirradiation in the present study in mice is similar to our previous finding in cultured cell system. Histological study showed that the preirradiation suppressed both p53 and Bax accumulation in the spleen 7 days after challenging irradiation with 3 Gy. From these results, the preirradiation seems to decrease apoptosis after challenging high-dose irradiation, which would be induced by p53 and Bax signal transduction. Decreased apoptosis would result in an increase in cell survival of the spleen, a hematopoietic tissue. This might favor recovery from radiation-induced acute hematopoietic injury, and finally increased survival rate. This situation seems to be induced through apoptosis of highly radiation-sensitive cells after the priming irradiation that leaves resistant cells: the radiation-sensitive cells would be replaced during the interval time by radiation-resistant cells

  13. Effects of acupoint versus non-acupoint electroacupuncture on cerebral cortical neuronal Bcl-2,Bax and caspase-3 expression in a rat model of focal cerebral ischemia

    Jun Wang; Junming Fan; Yongshu Dong; Xia Huang; Hongxia Zhang

    2008-01-01

    BACKGROUND: Several studies have demonstrated that electroacupuncture by acupoint selection can inhibit cerebral cortical neuronal apoptosis following cerebral ischemia/reperfusion.OBJECTIVE: To validate the effects of electroacupuncture by acupoint selection on the expression level of cortical neuronal anti-apoptotic Bcl-2 protein and the apoptotic executive protein, caspase-3, in rat models of focal cerebral ischemia/reperfusion.DESIGN, TIME AND SETTING: This randomized grouping, neural cell and molecular biology animal experiment was performed at the Laboratory of Pharmacology of Traditional Chinese Medicine and the Laboratory Animal Center of Henan Institute of Traditional Chinese Medicine between November 2006 and May 2007.MATERIALS: Atotal of 40 healthy male adult Sprague-Dawley rats were randomly and evenly divided into four groups: sham-operated, model, electroacupuncture and non-acupoint control. G6895 electro-acupuncture instruments were purchased from Shanghai Huayi Instrument Factory, China. Caspase-3, Bcl-2 and Bax kits were provided by Wuhan Boster Bioengineering Co., Ltd., China.METHODS: Middle cerebral artery occlusion was induced in the model, electroacupuncture and non-acupoint groups. In the electroacupuncture group, the acupoints Jianyu (LI15), Waiguan (SJ5), Biguan (ST31), and Zusanli (ST36) were given electroacupuncture. In the non-acupoint control group, at each time point (immediately after ischemia and after reperfusion, or 2 hours after reperfusion), electroacupuncture was performed at the midpoints of Tianquan (PC2)-Quze (PC 3) line, Quze (PC 3)-Ximen (PC4) line, Zuwuli (LRlO)-Yinbao (LRg) line, and Xiguan (LR7)-Zhongdu (LR6) line. Electroacupuncture parameters were set with a continuous wave with a frequency of 10 Hz, wave width 0.6 ms, voltage 1.5-3.0 V, and a duration of 10 minutes. The sham-operated and model groups received only animal fixation without electroacupuncture procedure.MAIN OUTCOME MEASURES: Five rats were selected from

  14. Preparation of BaxSr1-xTiO3 Functional Ceramic Film by Liquid Self-Assembly Technology%液相自组装制备BaxSr1-xTiO3功能陶瓷薄膜

    谈国强; 程蕾; 苗鸿雁; 王艳

    2011-01-01

    以Sr(NO3)2,Ba(NO3)2,(NH4)2TiF6和H3BO3作为原料,采用自组装单分子膜(self-assembled monolayers,SAMs)技术以及利用紫外光修饰技术对十八烷基三氯硅烷(C18H37SiCl3,OTS)单分子膜进行官能团改性,在氧化铟锡(indium tin oxide,ITO)玻璃基板上成功制备了BaxSr1-xTiO3功能陶瓷薄膜.通过动态/静态接触角仪测量了SAMs功能化的ITO玻璃基板表面与水的接触角,探讨功能化ITO玻璃基板在紫外光照射前后的润湿情况.通过X射线衍射、能量色散光谱和扫描电子显微镜等测试方法分析了制备的BaxSr1-xTiO3功能陶瓷薄膜的物相组成、微区结构和形貌.结果表明:改性的SAMs功能化ITO玻璃基板在50℃的前驱溶液中沉积18h后,在600℃煅烧晶化2h,可以成功制备出纯相BaxSr1-xTiO3功能陶瓷薄膜,薄膜的颗粒均一,形貌均匀.%Using strontium nitrate, barium nitrate, ammonium hexafluorotitanate and boric acid as raw materials, the functional groups of octadecyltrichlorosilane (C18H37SiCl3, OTS) monolayer were modified via self-assembled monolayers (SAMs) technology and ultraviolet modification technique, and the BaxSr1-xTiO3 functional ceramic film on the indium tin oxide (ITO) glass substrate was prepared successfully. The contact angle between the surface of the SAM functionalized ITO glass substrate and water was measured by dynamic/static contact angle instrument in order to study the wetting conditions of the substrates before and after ultraviolet light irradiation. The phase composition, microstructure and morphology of the prepared BaxSr1-xTiO3 functional ceramic thin films were analyzed by X-ray diffraction, energy dispersive spectroscopy and scanning electron microscopy. The results indicate that pure phase BaxSr1-xTiO3 functional ceramic film can be prepared on the SAMs functionalized ITO glass substrate successfully after depositing on the surface of ITO glass substrate in the precursor at 50 ℃ for 18h, and then crystallized

  15. Both p53-PUMA/NOXA-Bax-mitochondrion and p53-p21cip1 pathways are involved in the CDglyTK-mediated tumor cell suppression

    Yu, Zhendong, E-mail: zdyu@hotmail.com [Department of Clinical laboratory, Peking University Shenzhen Hospital, Guangdong (China); Wang, Hao [Department of pathology, The Chinese University of Hong Kong, Hong Kong (China); Zhang, Libin; Tang, Aifa; Zhai, Qinna; Wen, Jianxiang; Yao, Li [Department of Clinical laboratory, Peking University Shenzhen Hospital, Guangdong (China); Li, Pengfei, E-mail: lipengfei@cuhk.edu.hk [Department of pathology, The Chinese University of Hong Kong, Hong Kong (China)

    2009-09-04

    CDglyTK fusion suicide gene has been well characterized to effectively kill tumor cells. However, the exact mechanism and downstream target genes are not fully understood. In our study, we found that CDglyTK/prodrug treatment works more efficiently in p53 wild-type (HONE1) cells than in p53 mutant (CNE1) cells. We then used adenovirus-mediated gene delivery system to either knockdown or overexpress p53 and its target genes in these cells. Consistent results showed that both p53-PUMA/NOXA/Bcl2-Bax and p53-p21 pathways contribute to the CDglyTK induced tumor cell suppression. Our work for the first time addressed the role of p53 related genes in the CDglyTK/prodrug system.

  16. Diffuse phase transition and high-temperature dielectric relaxation study on (Bi0.5Na0.5)1-xBaxTiO3 ceramics

    Chen, Feng; Liu, Qiu-Xiang; Tang, Xin-Gui; Jiang, Yan-Ping; Yue, Jing-Long; Li, Jin-Kai

    2016-09-01

    Lead free (Bi0.5Na0.5)1-xBaxTiO3 (x=0.6, 0.7, 0.8, 0.9) ferroelectric ceramics were synthesized by the traditional solid state reaction method. Sintering was carried out at 1200 °C for 2 h in air atmosphere. The structural, microstructure and Ferroelectric of ceramics were investigated. In dielectric studies, a diffuse phase transition was exhibited and a dielectric relaxation behavious was observed at high temperature range. Impedance analysis characterized grain and grain boundaries resistivities of the ceramics and calculated activation energy and the activation energy for conduction. Polaron theory indicates that the relaxation of the samples at high temperatures was associated with the hopping ions caused by oxygen vacancies.

  17. Inhibition of nuclear factor-kappaB or Bax prevents endoplasmic reticulum stress- but not nitric oxide-mediated apoptosis in INS-1E cells

    Tonnesen, Morten F; Grunnet, Lars G; Friberg, Josefine; Cardozo, Alessandra K; Billestrup, Nils; Eizirik, Décio L; Størling, Joachim; Mandrup-Poulsen, Thomas

    2009-01-01

    elicited by NO and ER Ca(2+) depletion differ, we here compare the direct effects of NO, in the form of the NO donor S-nitroso-N-acetyl-D,L-penicillamine (SNAP), with the effects of SERCA2 inhibitor thapsigargin (TG) on MAPK, nuclear factor kappaB (NFkappaB), Bcl-2 proteins, ER stress, and apoptosis...... and alternative splicing of the transcription factor Xbp-1 were exclusively activated by TG. TG exposure caused NFkappaB activation, as assessed by IkappaB degradation and NFkappaB DNA binding. Inhibition of NFkappaB or the Bcl-2 family member Bax pathways protected beta-cells against TG- but not SNAP...

  18. Both p53-PUMA/NOXA-Bax-mitochondrion and p53-p21cip1 pathways are involved in the CDglyTK-mediated tumor cell suppression

    CDglyTK fusion suicide gene has been well characterized to effectively kill tumor cells. However, the exact mechanism and downstream target genes are not fully understood. In our study, we found that CDglyTK/prodrug treatment works more efficiently in p53 wild-type (HONE1) cells than in p53 mutant (CNE1) cells. We then used adenovirus-mediated gene delivery system to either knockdown or overexpress p53 and its target genes in these cells. Consistent results showed that both p53-PUMA/NOXA/Bcl2-Bax and p53-p21 pathways contribute to the CDglyTK induced tumor cell suppression. Our work for the first time addressed the role of p53 related genes in the CDglyTK/prodrug system.

  19. Dielectric dispersion of BaxSr1-xTiO3 thin film with parallel-plate and coplanar interdigital electrodes

    Ferroelectric BaxSr1-xTiO3 (BST) thin films with x = 0.25 and 0.5 were grown by pulsed laser deposition on single crystal LaAlO3 and Pt/Ti/SiO2/Si substrates, respectively. Capacitors were then fabricated from the BST thin films based on coplanar interdigital electrodes (CIEs) and parallel-plate electrodes (PPEs). The dielectric properties of the BST film with CIE and PPE were investigated and compared over a wide frequency range from 100 Hz to 10 GHz. The dielectric dispersion in PPE configuration, caused by the interfacial polarization in film/electrode interfaces, exhibited a strong dependence on frequency. However, the permittivity εCIE in CIE configuration shows a gentle variation with the frequency indicating interfacial polarization substantially suppressed. The influence upon the dielectric properties of the columnar BST grains due to the use of different forms of electrodes was discussed.

  20. Characteristics of silicon-based BaxSr1-xTiO3 thin films prepared by a sol-gel method

    Silicon-based BaxSr1-xTiO3 (BST) thin films have been prepared by a sol-gel method with rapid thermal annealing (RTA) processes. Phase structure of the films has been investigated by x-ray diffraction. Atomic force microscopy studies reveal a dense and smooth surface of the sol-gel prepared films. Microstructure and electrical properties of the BST films can be affected by the substrate and the annealing process. RTA method is found to be very efficient to improve the electrical properties of the films. Dielectric constant and dielectric loss of the BST films at 100 kHz are 230 and 0.02, respectively. Leakage current density of the BST capacitors is 1.6x10-7A cm-2 at 3 V. (author)

  1. AC Conductivity and Dielectric Relaxation Behavior of Sol-gel BaxSr1-xTiO3 Thin Films

    Ala'eddin A. Saif; P. Poopalan

    2011-01-01

    BaxSr1-xTiO3 sol-gel thin films with x--0.5, 0.7 and 0.8 have been fabricated as AI/BST/Pt capacitor. The AC conductivity and dielectric properties over a frequency rang of 10 Hz and I MHz have been studied in order to explore the ion dynamics and relaxation mechanisms in the films. The frequency dependent conductivity plots show three regions of conduction processes. Dielectric results show that ε' at low frequencies increases as Sr content decreases, whereas at high frequencies, it shows opposite variation, which is attributed to the dipole dynamics. The electric modulus plots reveal the relaxation peaks which are not observed in the ε" plots and the contribution of the grains, grain boundaries and electrode to the relaxation mechanisms.

  2. Selective cytotoxicity of squamocin on T24 bladder cancer cells at the S-phase via a Bax-, Bad-, and caspase-3-related pathways.

    Yuan, Shyng-Shiou F; Chang, Hsueh-Ling; Chen, Hsiao-Wen; Kuo, Fu-Chen; Liaw, Chih-Chuang; Su, Jinu-Huang; Wu, Yang-Chang

    2006-01-18

    Annonaceous acetogenins are a group of potential anti-neoplastic agents isolated from Annonaceae plants. We purified squamocin, a cytotoxic bis-tetrahydrofuran acetogenin, from the seeds of Annona reticulata and analyzed its biologic effects on cancer cells. We showed that squamocin was cytotoxic to all the cancer lines tested. Furthermore, squamocin arrested T24 bladder cancer cells at the G1 phase and caused a selective cytotoxicity on S-phase-enriched T24 cells. It induced the expression of Bax and Bad pro-apoptotic genes, enhanced caspase-3 activity, cleaved the functional protein of PARP and caused cell apoptosis. These results suggest that squamocin is a potentially promising anticancer compound. PMID:16154156

  3. Annonacin, a mono-tetrahydrofuran acetogenin, arrests cancer cells at the G1 phase and causes cytotoxicity in a Bax- and caspase-3-related pathway.

    Yuan, Shyng-Shiou F; Chang, Hsueh-Ling; Chen, Hsiao-Wen; Yeh, Yao-Tsung; Kao, Ying-Hsien; Lin, Kuei-Hsiang; Wu, Yang-Chang; Su, Jinu-Huang

    2003-05-01

    Annonaceous acetogenins are a group of potential anti-neoplastic agents isolated from Annonaceae plants. In this study, we purified annonacin, a cytotoxic mono-tetrahydrofuran acetogenin, from the seeds of Annona reticulata and analyzed its biological effects. Herein, we have shown that annonacin caused significant cell death in various cancer cell lines. T24 bladder cancer cells at the S phase were more vulnerable to the cytotoxicity of annonacin. Furthermore, annonacin activated p21 in a p53-independent manner and arrested T24 cells at the G1 phase. It also induced Bax expression, enhanced caspase-3 activity, and caused apoptotic cell death in T24 cells. In summary, these results suggest that annonacin is potentially a promising anti-cancer compound. PMID:12697268

  4. cAMP signalling of Bordetella adenylate cyclase toxin through the SHP-1 phosphatase activates the BimEL-Bax pro-apoptotic cascade in phagocytes.

    Ahmad, Jawid Nazir; Cerny, Ondrej; Linhartova, Irena; Masin, Jiri; Osicka, Radim; Sebo, Peter

    2016-03-01

    The adenylate cyclase toxin-hemolysin (CyaA, ACT or AC-Hly) plays a key role in virulence of Bordetella pertussis. CyaA penetrates myeloid cells expressing the complement receptor 3 (αM β2 integrin CD11b/CD18) and subverts bactericidal capacities of neutrophils and macrophages by catalysing unregulated conversion of cytosolic ATP to the key signalling molecule adenosine 3',5'-cyclic monophosphate (cAMP). We show that the signalling of CyaA-produced cAMP hijacks, by an as yet unknown mechanism, the activity of the tyrosine phosphatase SHP-1 and activates the pro-apoptotic BimEL-Bax cascade. Mitochondrial hyperpolarization occurred in human THP-1 macrophages within 10 min of exposure to low CyaA concentrations (e.g. 20 ng ml(-1) ) and was accompanied by accumulation of BimEL and association of the pro-apoptotic factor Bax with mitochondria. BimEL accumulation required cAMP/protein kinase A signalling, depended on SHP-1 activity and was selectively inhibited upon small interfering RNA knockdown of SHP-1 but not of the SHP-2 phosphatase. Moreover, signalling of CyaA-produced cAMP inhibited the AKT/protein kinase B pro-survival cascade, enhancing activity of the FoxO3a transcription factor and inducing Bim transcription. Synergy of FoxO3a activation with SHP-1 hijacking thus enables the toxin to rapidly trigger a persistent accumulation of BimEL, thereby activating the pro-apoptotic programme of macrophages and subverting the innate immunity of the host. PMID:26334669

  5. Activation of A2b adenosine receptor regulates ovarian cancer cell growth: involvement of Bax/Bcl-2 and caspase-3.

    Hajiahmadi, Sima; Panjehpour, Mojtaba; Aghaei, Mahmoud; Shabani, Mahdi

    2015-08-01

    A2b adenosine receptor (A2bAR) acts as a potent regulator of cell growth in various cell lines. The present study was designed to understand the controlling mechanism of A2bAR agonist (NECA)-induced apoptosis in ovarian cancer cells. Real-time PCR and western blotting assays were used to evaluate the gene and protein expression profiles of A2bAR, respectively. MTT assay was used to study the cell proliferation effect of A2bAR agonist (NECA). Detection of apoptosis was conducted using annexin V-FITC/PI staining, caspase-3 activation assay, and the expression of Bax and Bcl-2 proteins analysis. The mitochondrial membrane potential (ΔΨM) was analyzed by employing JC-1 prob. The mRNA and protein expression levels of A2bAR in ovarian cancer cells were detected. NECA significantly reduced cell viability in a dose-dependent manner in OVCAR-3 and Caov-4 cell lines. The growth inhibition effect of NECA was related to the induction of cell apoptosis, which was manifested by annexin V-FITC staining, activation of caspase-3, and loss of mitochondrial membrane potentials (ΔΨm). In addition, downregulation of the regulatory protein Bcl-2 and upregulation of Bax protein by NECA were also observed. These findings demonstrated that NECA induces apoptosis via the mitochondrial signaling pathway. Thus, A2bAR agonists may be a potential agent for induction of apoptosis in ovarian cancer cells. PMID:25877700

  6. The ethanol extract of Scutellaria baicalensis and the active compounds induce cell cycle arrest and apoptosis including upregulation of p53 and Bax in human lung cancer cells

    Despite a lack of scientific authentication, Scutellaria baicalensis is clinically used in Chinese medicine as a traditional adjuvant to chemotherapy of lung cancer. In this study, cytotoxicity assays demonstrated that crude ethanolic extracts of S. baicalensis were selectively toxic to human lung cancer cell lines A549, SK-LU-1 and SK-MES-1 compared with normal human lung fibroblasts. The active compounds baicalin, baicalein and wogonin did not exhibit such selectivity. Following exposure to the crude extracts, cellular protein expression in the cancer cell lines was assessed using 2D gel electrophoresis coupled with MALDI-TOF-MS/Protein Fingerprinting. The altered protein expression indicated that cell growth arrest and apoptosis were potential mechanisms of cytotoxicity. These observations were supported by PI staining cell cycle analysis using flow cytometry and Annexin-V apoptotic analysis by fluorescence microscopy of cancer cells treated with the crude extract and pure active compounds. Moreover, specific immunoblotting identification showed the decreased expression of cyclin A results in the S phase arrest of A549 whereas the G0/G1 phase arrest in SK-MES-1 cells results from the decreased expression of cyclin D1. Following treatment, increased expression in the cancer cells of key proteins related to the enhancement of apoptosis was observed for p53 and Bax. These results provide further insight into the molecular mechanisms underlying the clinical use of this herb as an adjuvant to lung cancer therapy. - Research highlights: → Scutellaria baicalensis is a clinical adjuvant to lung cancer chemotherapy in China. → Scutellaria ethanol extracts selectively toxic to A549, SK-LU-1 and SK-MES-1. → Baicalin, baicalein and wogonin were toxic to all lung cancer cell lines. → Proteomics identified increased p53 and BAX in response to Scutellaria extracts.

  7. Effects of β-Estradiol on the level of Bcl2/BAX Expression in the Rat Hippocampus, after Induction of Post-Traumatic Stress Disorder (PTSD

    Shaghayegh Saffari

    2015-01-01

    Full Text Available Introduction: Post-traumatic stress disorder (PTSD is an anxiety disorder, that can occur in people who have experienced or witnessed a natural disaster, serious accident or other life-threatening events. It's most important characteristics are memory disorders and hippocampus is one of the essential structure in this relation. Traumatic events cause apoptosis and alter the expression of neurotrophic factors in hippocampus. Therefore, the aim of this study was the examination of the fear memory and evaluation the effects of multiple β-Estradiol injections on behavioral responses in the rat hippocampus, after induction of PTSD. Moreover, we evaluated, the biochemical and histological mechanisms of β-Estradiol on hippocampus. Methods: We used single prolonged stress (SPS, for PTSD induction. One day after SPS, rats received electrical foot shock within shock chamber. To test the conditioned fear responses one week later, rats were placed back in the chamber without any shock, and freezing behavior was defined. Another fear test was done 4 days after the first test. Animals received multiple subcutaneous injections of β-estradiol (45µg/kg or sesame oil, immediately after shock and in these 7 days, between shock and test. Bcl-2 and BAX were detected by immunohistochemistry. Results: Main findings of the present study are: Exaggerated fear response was seen in PTSD group as compared with control group. Moreover, Β-estradiol administration reduced behavioral responses, caused by PTSD induction. SPS decreased the Bcl2/BAX ratio and apoptosis induced by SPS in hippocampus. This effects reversed by β-estradiol injections. Conclusion: β-estradiol could be useful for treating the memory disorders in PTSD.

  8. Adsorption of xylene para- and meta- isomers in NaX and BaX zeolites. Study of properties-structure relations; Adsorption des isomeres para- et meta- du xylene dans les zeolithes NaX et BaX. Etude des relations proprietes-structure

    Descours, A.

    1997-02-14

    The separation of para-xylene from C8 aromatics is performed industrially bu adsorption process on zeolitic molecular sieves. The sorption properties of these zeolites are strongly linked to their structure, and their comprehension require an accurate knowledge of the interactions between sorbate molecules and zeolitic structure. The aim of this work is to characterise from a structural point of view the adsorption of para- and meta-xylenes in BaX and NaX zeolites. The former is selective for para-xylene, and the latter has not selective properties for para- and meta-isomers of xylene. For each zeolite, the adsorption of pure para-xylene and meta-xylene or a mixture of the two isomers, is investigated as a function of coverage. Powder neutron diffraction is used to determine the crystalline structure of these zeolites and the different crystallographic adsorption sites of the molecules. The influence of coverage on sorbate-sorbent and sorbate-sorbate interactions is investigated. Infrared spectroscopy allows to determine the chemical environment of the sorbate molecules at low coverage or when the coverage increases, and is particularly effective for the study of the binary mixture of xylenes. This study is performed by sorbing a mixture of xylene isomers, or by sorbing these isomers successively. Infrared studies and crystallographic analysis are compared in order to get a consistent description of adsorption mechanism of xylene isomers for both zeolites as a function of coverage. The role of coverage, of cation type, an the presence of the two xylene isomers is the super-cages is essential. For both zeolites, the increase of coverage actually leads to steric hindrances between sorbed molecules and molecular rearrangements. These reorganizations are connected to the cationic distribution of NaX and BaX zeolites. The sorbed molecules are connected to the cationic distribution of NaX and BaX zeolites. The sorbed molecules are particularly confined in BaX zeolite

  9. Ginkgo biloba extract mitigates liver fibrosis and apoptosis by regulating p38 MAPK, NF-κB/IκBα, and Bcl-2/Bax signaling

    Wang YY

    2015-12-01

    Full Text Available Yuanyuan Wang, Rong Wang, Yujie Wang, Ruqin Peng, Yan Wu, Yongfang Yuan Department of Pharmacy, Shanghai 9th People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, People’s Republic of China Background: Liver fibrosis is the consequence of diverse liver injuries and can eventually develop into liver cirrhosis. Ginkgo biloba extract (GBE is an extract from dried ginkgo leaves that has many pharmacological effects because of its various ingredients and has been shown to be hepatoprotective. Purpose and methods: Aimed to investigate the underlying protective mechanisms of GBE on carbon tetrachloride (CCl4-induced liver fibrosis in rats. Male Sprague Dawley rats were randomly divided into four groups: control group (C, model group (M, low-dose group (L, and high-dose group (H. Liver fibrosis was induced by CCl4 groups M, L, and H: group C was administered saline. In addition, GBE at different doses was used to treat groups L and H. Results: The results of hematoxylin and eosin staining, Masson’s trichrome staining, a liver function index, and a liver fibrosis index showed that GBE application noticeably mitigated fibrosis and improved the function of the liver. The western blotting and immunohistochemistry analyses indicated that GBE reduced liver fibrosis not only by inhibiting p38 MAPK and NF-κBp65 via inhibition of IκBα degradation but also by inhibiting hepatocyte apoptosis via downregulation of Bax, upregulation of Bcl-2, and subsequent inhibition of caspase-3 activation. Inflammation-associated factors and hepatic stellate cell (HSC-activation markers further demonstrated that GBE could effectively inhibit HSC activation and inflammation as a result of its regulation of p38 MAPK and nuclear factor-kappa B/IκBα signaling. Conclusion: Our findings indicated a novel role for GBE in the treatment of liver fibrosis. The potential mechanisms may be associated with the following signaling pathways: 1 the p38 MAPK

  10. Expression of P53, P21/WAF/CIP, BCL-2, BAX, BCL-X, and BAK in radiation-induced apoptosis in testicular germ cell tumor lines

    Purpose: Testicular germ cell tumors (TGCTs) represent one of the few tumor types that are curable by antineoplastic therapy, probably due to the high sensitivity of this neoplasm to induction of apoptosis by chemotherapeutic agents and/or ionizing radiation. Here, we tested cell susceptibility to radiation-induced apoptosis in a panel of TGCT cell lines and attempted to correlate this with the known potentially relevant molecular determinants (p53 gene status and Bcl-2 family proteins) of apoptosis. Methods and Materials: Induction of apoptosis by γ-radiation was morphologically recognized in NT2, NCCIT, S2, and 2102 EP using Hoechst/PI staining and additionally confirmed by Western blot analysis of PARP cleavage. The p53 gene status was estimated by sequence analysis. Expression of p21/WAF/CIP was determined by Northern blot analysis and immunoblotting was used to monitor p53, Bax, Bcl-2, Bcl-x, and Bak protein levels. In vitro colony formation was studied to establish clonogenic survival curves. Results: NT2 and NCCIT appeared to be susceptible for radiation-induced apoptosis, contrasting 2102 EP and S2 which were highly resistant. Sequence analysis showed that NT2, S2, and 2102 EP are homozygous for wild-type p53 (wtp53), whereas NCCIT contains mutant p53 (mtp53). NT2 and 2102 EP cells showed radiation-induced p53 upregulation, while NCCIT (mtp53) and S2 (no p53 protein) cells did not. Consistently, γ-radiation-induced DNA damage resulted in a p53-dependent transactivation of the p21/WAF/CIP gene in NT2 and 2102 EP, but not in mtp53-containing NCCIT cells and p53 nonexpressing S2 cells. Constitutive expression of Bax, Bcl-2, Bcl-x, and Bak was not affected by radiation and showed no correlation with cell susceptibility to radiation-induced apoptosis. A discrepancy was found between apoptosis and reproductive death. Conclusions: The present study revealed that: i) the presence of wtp53 may not be absolutely required for the hypersensitivity for radiation

  11. Gastroprotective activity of Annona muricata leaves against ethanol-induced gastric injury in rats via Hsp70/Bax involvement

    Moghadamtousi SZ

    2014-10-01

    downregulation of Bax and upregulation of Hsp70 proteins after pretreatment. Collectively, the present results suggest that EEAM has a promising antiulcer potential, which could be attributed to its suppressive effect against oxidative damage and preservative effect toward gastric wall mucus. Keywords: Annona muricata, annonaceae, gastric injury, antioxidants, Hsp70/Bax

  12. 血糖波动对2型糖尿病大鼠认知功能及海马Bax/Bcl-2的影响%The Effect of Glucose Fluctuate on the Cognitive Function and the Bax/Bcl-2 in Hippocampal of Rats with Type 2 Diabetes Mellitus

    徐志伟; 寿旗扬; 李守业; 蔡月琴; 刘琼; 徐文聃; 洪春兰; 王辉

    2013-01-01

    [目的]探讨血糖波动对糖尿病大鼠认知功能和海马Bax/Bcl-2的影响.[方法]取45只雄性大鼠禁食不禁水16h,用链脲佐菌素30 mg·kg-1加高脂饲料诱导2型糖尿病大鼠模型,挑选出糖尿病大鼠30只,分成糖尿病模型组(Diabetes Model,M)、持续高血糖糖尿病组(High Glucose Diabetes Sustain Model,MS)、血糖波动糖尿病组(Glucose Fluctuate Diabetes Model,MF),另取10只正常大鼠做正常对照组(Normal,N).血糖波动糖尿病组错时腹腔注射葡萄糖0.38 g·kg-1和皮下注射胰岛素1U制备糖尿病血糖波动模型.血糖波动6周后,进行水迷宫定位航行实验和空间探索实验,同时检测海马组织中Bax、Bcl-2的表达.[结果]M组、MS组、MF组比N组逃避潜伏期明显增长(P<0.05),空间探索实验结果显示60s内穿越平台所在位置的次数明显下降,在平台所在象限内的游泳距离明显减少(P<0.05).MF组和M组、MS组相比.逃避潜伏期进一步增长(P<0.05),空间探索实验结果显示60s内穿越平台所在位置次数也明显下降,在平台所在象限游泳距离也减少(P<0.05).MF组、M纽、MS组Bcl-2的表达较N组显著下降,Bax显著升高(P<0.05).MF组Bcl-2的表达较M组、MS组进一步下降,Bax进一步升高(P<0 05).[结论]血糖波动能加重损伤糖尿病大鼠的学习记忆功能,并促进海马组织的凋亡相关蛋白Bax/Bcl-2的表达,诱导海马神经元的凋亡.%[Objective] To explore the effection of glucose fluctuation on the cognitive function and the Bax/Bcl-2 in hippocampal of rats with type 2 diabetes mellitus. [Methods] The SD rats were randomly divided into four groups: normal group(N); diabetes model group(M);high glucose sustain diabetes model group(MS); glucose fluctuate diabetes model group(MF). The diabetic rats model was developed by injection with streptozotocin(STZ) 30 mg·kg-1, regularly staggered insulin 1U and glucose 0.38 mg·kg-1 administration was used to set up blood

  13. Enhancement of Photon Absorption on BaxSr1-xTiO3 Thin-Film Semiconductor Using Photonic Crystal

    Abd. Wahidin Nuayi

    2014-01-01

    Full Text Available Enhancement of photon absorption on barium strontium titanate (BaxSr1-xTiO3 thin-film semiconductor for mole fraction x=0.25, 0.35, 0.45, and 0.55 using one-dimensional photonic crystal with defect was investigated experimentally. The thin film was grown on transparent conductive oxide (TCO substrate using chemical solution deposition method and annealed at 500°C for 15 hours with increasing rate of 1.6°C/min. From optical characterization in visible spectrum it was found that the average absorption percentages are 92.04%, 83.55%, 91.16%, and 80.12%, respectively. The BST thin film with embedded photonic crystal exhibited a relatively significant enhancement on photon absorption, with increasing value of 3.96%, 7.07%, 3.04%, and 13.33% for the respective mole fraction and demonstrating absorbance characteristic with flat feature. In addition, we also discuss the thin-film properties of attenuation constant and electrical conductivity.

  14. Bax Inhibitor-1, a Conserved Cell Death Suppressor, Is a Key Molecular Switch Downstream from a Variety of Biotic and Abiotic Stress Signals in Plants

    Naohide Watanabe

    2009-07-01

    Full Text Available In Nature plants are constantly challenged by a variety of environmental stresses that could lead to disruptions in cellular homeostasis. Programmed cell death (PCD is a fundamental cellular process that is often associated with defense responses to pathogens, during development and in response to abiotic stresses in fungi, animals and plants. Although there are many characteristics shared between different types of PCD events, it remains unknown whether a common mechanism drives various types of PCD in eukaryotes. One candidate regulator for such a mechanism is Bax Inhibitor-1 (BI-1, an evolutionary conserved, endoplasmic reticulum (ER-resident protein that represents an ancient cell death regulator that potentially regulates PCD in all eukaryotes. Recent findings strongly suggested that BI-1 plays an important role in the conserved ER stress response pathway to modulate cell death induction in response to multiple types of cell death signals. As ER stress signaling pathways has been suggested to play important roles not only in the control of ER homeostasis but also in other biological processes such as the response to pathogens and abiotic stress in plants, BI-1 might function to control the convergence point that modulates the level of the “pro-survival and pro-death” signals under multiple stress conditions.

  15. Layered Bi4BaxV2-xO11-(3x/2)-δ perovskite oxide as solid electrolyte for intermediate temperature solid oxide fuel cells

    A continuous series of Bi4BaxV2-xO11-(3x/2)-δ solid solutions are prepared by conventional solid state reactions. The polymorphism and electrical properties of these samples are studied by FT-IR spectroscopy, X-ray diffraction, differential thermal analysis (DTA) and AC impedance. The solid solutions with composition 0.07≤x≤0.13 are isostructural with the monoclinic phase α-Bi4V2O11. However, orthorhombic β-phase is observed for x=0.17 and tetragonal γ-phase is stabilized for x≤0.30. X-ray and DTA results reveal the occurrence of α↔γ transition for x≤0.13, β↔γ transition for x=0.17 and γ'→γ transition for x≥0.20. AC impedance plots at 280 deg. C for all compositions show greater contribution of grain to ionic conductivity than grain boundary. The highest ionic conductivity, σ300oC=4.456x10-5 S cm-1 is observed for x=0.17. The ionic conductivity of the substituted compounds is higher than the parent α-Bi4V2O11, due to the increased oxygen ion vacancies generated as a result of barium doping.

  16. Preparation and Dielectric Properties of (BaxSr1-xTiO3/Mg2TiO4 Composite Ceramics withA Functionally Graded Structure

    LI Jun, WANG Xu-Sheng, CHAI Xiao Na, LIU Peng

    2014-01-01

    Full Text Available (BaxSr1-xTiO3/Mg2TiO4 composite ceramics with a constituent graded structure were prepared by a solid-state reaction method. Their microstructure and dielectric properties were analyzed by X-Ray diffraction (XRD, Scanning Electron Microscope (SEM, Energy Dispersive Spectroscope (EDS and dielectric property measurements. The results show that the samples sintered at 1375°C for 3 h are a composite phase of perovskite and spinel structure with a Ba/Sr ratio composition gradient. Compared with the composite samples with a fixed Ba/Sr ratio, the graded samples possess higher tunability and better temperature stability of dielectric properties. At room temperature (20°C, the tunability of a typical sample is 21.9% under an external DC field of 2 kV/mm, and it maintains a high value of 9.3% at high temperature of 60°C. The improvement of temperature stability is due to the difference in Curie temperature for different layers in gradient (Ba,SrTiO3. Meanwhile, the introduction of the composition gradient in this material helps us to obtain a wide temperature application range for the related device.

  17. Electrical properties and water incorporation in A-site deficient perovskite La1-xBaxNb3O9-0.5x

    Animitsa, I.; Iakovleva, A.; Belova, K.

    2016-06-01

    Barium doped A-site deficient perovskites La1-xBaxNb3O9-0.5x (x=0-0.05) were synthesized by the solid state method, their structure, electrical properties and state of oxygen-hydrogen groups have been investigated. These phases were found to be able to incorporate water from the gas phase and to exhibit proton transport. Hydration is accompanied by the formation of different forms of oxygen-hydrogen groups: OH- - groups and H3O+ - ions. The total conductivities of doped samples increased in a wet atmosphere due to the appearance of proton current carriers (at the temperatures below 700 °C), but the conductivity increased insignificantly (~0.25 order of magnitude) because of a low doping level and, consequently, small concentration of protons. TG-measurements confirmed relatively low water content (below 0.2%). The total conductivity depends substantially on x and exhibits a minimum on σ-f(x) dependencies. It has been suggested that such behavior is a manifestation of a mixed cation effect.

  18. Bcl-2、Bax及M30在Bowen病中的表达%Expression of Bcl-2、 Bax and M30 in Bowen's disease

    张敏; 张谊之; 王琳; 李俸媛

    2002-01-01

    为了研究细胞凋亡与Bowen病发病的关系,我们采用免疫组化方法检测了28例Bowen病皮损中Bcl-2、Bax及M30的表达.结果发现M30及Bax染色阳性分别见于4例和8例表皮角质形成细胞,Bcl-2在10例表皮角质形成细胞和12例真皮淋巴细胞呈阳性表达;Bcl-2表达阳性率高于Bax,但无统计学差异(χ2=2.1053,P>0.05);M30与Bax表达显著正相关(r=0.6455,P0.05).提示Bowen病发病可能与表皮角质形成细胞凋亡降低有关.

  19. Moessbauer study of the relationship between magnetic properties and Jahn-Teller distortion in La1-xBax(Mn,Fe)O3 perovskites

    The local structural distortion and its effect on the magnetic and magnetotransport properties of the Fe-doped La1-xBaxMnO3 (0.00≤x≤0.35) compounds were investigated by means of X-ray diffraction, Moessbauer spectroscopy, and magnetization measurements. The Moessbauer spectra show clear evidence of the local structural distortion of the Mn(Fe)O6 octahedron on the basis of non-zero nuclear quadrupole interactions for high-spin Fe3+ ions. It was found that the local structural distortion decreases significantly with the introduction of a few per cent of Ba, reaches a minimum around x=0.13, and then increases slightly on further increasing the Ba concentration. The Ba-concentration dependence of the Jahn-Teller coupling strength estimated from the Moessbauer results was found to be consistent with the magnetic properties. Consequently, the connection between this local structural distortion and the colossal magnetoresistive behavior of this family of materials can be well explained. (copyright 2005 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim) (orig.)

  20. Phenomenological theory of phase transitions in epitaxial BaxSr1-xTiO3 thin films on (111)-oriented cubic substrates

    Shirokov, V. B.; Shakhovoy, R. A.; Razumnaya, A. G.; Yuzyuk, Yu. I.

    2015-07-01

    A phenomenological thermodynamic theory of BaxSr1-xTiO3 (BST-x) thin films epitaxially grown on (111)-oriented cubic substrates is developed using the Landau-Devonshire approach. The group-theoretical analysis of the low-symmetry phases was performed taking into account two order parameters: the polarization related to ionic shifts in polar zone-center F1u mode and the out-of-phase rotation of TiO6 octahedra corresponding to the R25 zone-boundary mode in the parent cubic phase P m 3 ¯ m . The eight-order thermodynamic potential for BST-x solid solutions was developed and analyzed. We constructed the "concentration-misfit strain" phase diagram for BST-x thin films at room temperature and found that polar rhombohedral R3m phase with the polarization normal to the substrate is stable for x > 0.72 and negative misfit strains, while ferroelectric monoclinic C2 and Cm phases with in-plane polarization are stable for much smaller x and positive or slightly negative misfit strains. We constructed the "temperature-misfit strain" phase diagrams for several concentrations (x = 1, 0.8, 0.6, 0.4, and 0.2). Systematic changes of the phase transition lines between the paraelectric and ferroelectric phases are discussed. The phase diagrams are useful for practical applications in thin-film engineering.

  1. Protective Effect of Aliskiren in Experimental Ischemic Stroke: Up-Regulated p-PI3K, p-AKT, Bcl-2 Expression, Attenuated Bax Expression.

    Miao, Jiangyong; Wang, Lina; Zhang, Xiangjian; Zhu, Chunhua; Cui, Lili; Ji, Hui; Liu, Ying; Wang, Xiaolu

    2016-09-01

    Aliskiren (ALK), a pharmacological renin inhibitor, is an effective antihypertensive drug and has potent anti-apoptotic activity, but it is currently unknown whether ALK is able to attenuate brain damage caused by acute cerebral ischemia independent of its blood pressure-lowering effects. This study aimed to investigate the role of ALK and its potential mechanism in cerebral ischemia. C57/BL6 mice were subjected to transient middle cerebral artery occlusion (tMCAO) and treated for 5 days with Vehicle or ALK (10 or 25 mg/kg per day via intragastric administration), whereas Sham-operated animals served as controls. Treatment with ALK significantly improved neurological deficits, infarct volume, brain water content and Nissl bodies after stroke (P < 0.05), which did not affect systemic blood pressure. Furthermore, the protection of ALK was also related to decreased levels of apoptosis in mice by enhanced activation of phosphatidylinositol 3-kinase (PI3K)/AKT pathway, increased level of Bcl-2 and reduced Bax expression (P < 0.05). In addition, ALK's effects were reversed by PI3K inhibitors LY294002 (P < 0.05). Our data indicated that ALK protected the brain from reperfusion injuries without affecting blood pressure, and this effect may be through PI3K/AKT signaling pathway. PMID:27180190

  2. Strain induced enhancement of magnetization in Ba2FeMoO6 based heterostructure with (BaxSr1-x)TiO3

    Kim, Kyeong-Won; Ghosh, Siddhartha; Buvaev, Sanal; Hebard, Arthur F.; Norton, David P.

    2016-05-01

    High quality epitaxial Ba2FeMoO6 thin films and Ba2FeMoO6-(BaxSr1-x)TiO3 bi-layer (BL) and superlattice (SL) structures were grown via pulsed laser deposition under low oxygen pressure, and their structural, magnetic, and magneto-transport properties were examined. Superlattice and bi-layer structures were confirmed by X-ray diffraction patterns. Low temperature magnetic measurement shows that the saturation magnetization (MS) is significantly higher for SLs and almost similar or lower for BLs, when compared to phase pure Ba2FeMoO6 thin films. The variation of the coercive field (HC) follows exact opposite trend, where BL samples have higher HC and SL samples have lower HC than pure Ba2FeMoO6 thin films. Also, a significant decrease of the Curie temperature is found in both BL and SL structures compared to pure Ba2FeMoO6 thin films. Negative magneto-resistance is seen in all the BL and SL structures as well as in pure Ba2FeMoO6 thin films. In contrast to the magnetic properties, the magneto-transport properties do not show much variation with induced strain.

  3. Lycopene modulates cholinergic dysfunction, Bcl-2/Bax balance, and antioxidant enzymes gene transcripts in monosodium glutamate (E621) induced neurotoxicity in a rat model.

    Sadek, Kadry; Abouzed, Tarek; Nasr, Sherif

    2016-04-01

    The effect of monosodium glutamate (MSG) on brain tissue and the relative ability of lycopene to avert these neurotoxic effects were investigated. Thirty-two male Wistar rats were distributed into 4 groups: group I, untreated (placebo); group II, injected with MSG (5 mg·kg(-1)) s.c.; group III, gastrogavaged with lycopene (10 mg·kg(-1)) p.o.; and group IV received MSG with lycopene with the same mentioned doses for 30 days. The results showed that MSG induced elevation in lipid peroxidation marker and perturbation in the antioxidant homeostasis and increased the levels of brain and serum cholinesterase (ChE), total creatine phosphokinase (CPK), creatine phosphokinase isoenzymes BB (CPK-BB), and lactate dehydrogenase (LDH). Glutathione S-transferase (GST), superoxide dismutase (SOD), and catalase (CAT) activities and gene expression were increased and glutathione content was reduced in the MSG-challenged rats, and these effects were ameliorated by lycopene. Furthermore, MSG induced apoptosis in brain tissues reflected in upregulation of pro-apoptotic Bax while lycopene upregulated the anti-apoptotic Bcl-2. Our results indicate that lycopene appears to be highly effective in relieving the toxic effects of MSG by inhibiting lipid peroxidation and inducing modifications in the activity of cholinesterase and antioxidant pathways. Interestingly, lycopene protects brain tissue by inhibiting apoptosis signaling induced by MSG. PMID:26900785

  4. DRP1-dependent apoptotic mitochondrial fission occurs independently of BAX, BAK and APAF1 to amplify cell death by BID and oxidative stress.

    Oettinghaus, Björn; D'Alonzo, Donato; Barbieri, Elisa; Restelli, Lisa Michelle; Savoia, Claudia; Licci, Maria; Tolnay, Markus; Frank, Stephan; Scorrano, Luca

    2016-08-01

    During apoptosis mitochondria undergo cristae remodeling and fragmentation, but how the latter relates to outer membrane permeabilization and downstream caspase activation is unclear. Here we show that the mitochondrial fission protein Dynamin Related Protein (Drp) 1 participates in cytochrome c release by selected intrinsic death stimuli. While Bax, Bak double deficient (DKO) and Apaf1(-/-) mouse embryonic fibroblasts (MEFs) were less susceptible to apoptosis by Bcl-2 family member BID, H2O2, staurosporine and thapsigargin, Drp1(-/-) MEFs were protected only from BID and H2O2. Resistance to cell death of Drp1(-/-) and DKO MEFs correlated with blunted cytochrome c release, whereas mitochondrial fragmentation occurred in all cell lines in response to all tested stimuli, indicating that other mechanisms accounted for the reduced cytochrome c release. Indeed, cristae remodeling was reduced in Drp1(-/-) cells, potentially explaining their resistance to apoptosis. Our results indicate that caspase-independent mitochondrial fission and Drp1-dependent cristae remodeling amplify apoptosis. This article is part of a Special Issue entitled 'EBEC 2016: 19th European Bioenergetics Conference, Riva del Garda, Italy, July 2-6, 2016', edited by Prof. Paolo Bernardi. PMID:26997499

  5. Study on the in-plane electrical resistivity and thermoelectric power in single crystals of La2-xBaxCuO4

    李鹏程; 杨宏顺; 李志权; 柴一晟; 曹烈兆

    2002-01-01

    The in-plane electrical resistivity and thermoelectric power have been measured on single crystals ofLa2-xBaxCuO4 at around x=0.125. The room temperature resistivity and thermopower have their maximum val-ues at x=0.125, indicating that the carrier concentration is the minimum and the carriers are most strongly localized atx=0.125. The observed semiconductor-like behaviour can be well described by the weak-localized quasi-two-dimensionalstate. The steep rise in electric resistivity of the sample at x=0.125 below 70K is attributed to the formation of staticstripe-order of holes and spins, which are pinned by the low-temperature tetragonal (LTT) structure, as discovered inLa1.4sNd0.4Sr0.12CuO4.The temperature dependence of electric resistivity below 70K is still well described by theformula p ∝ lnT. A definite change in the slope of thermopower is observed at the low-temperature orthorhombic-LTTstructural phase transition temperature. The origin of the 1/8 anomaly is discussed in the text.

  6. Composition and phase analysis of nanocrystalline BaxSr1-xFe12O19 (x = 1.0; 0.6; and 0.4) by using general structure analysis system

    Gunanto, Y. E.; Jobiliong, E.; Adi, Wisnu Ari

    2016-03-01

    Single phase of nanocrystalline BaxSr1-xFe12O19 (x = 1.0; 0.6; and 0.4) was successfully synthesized by mechanical milling method and thermal process. Stoichiometric quantities of analytical-grade SrCO3, BaCO3, and Fe2O3, were mixed and milled using a high-energy milling. The mixture of all precursors was sintered at a temperature of 1000 °C for 10 hours. The refinement of x-ray diffraction trace for all samples confirmed a single phase material with a hexagonal structure. The increase of the amount of strontium content in the barium atoms in the BaxSr1-xFe12O19 system can decrease the lattice parameter which have been successfully substituted into the barium atoms. The calculation result of cationic distribution showed that the BaxSr1-xFe12O19 (x = 0.6) and (x = 0.4) samples have nominal composition of Ba0,61Sr0,39Fe12O19 and Ba0,37Sr0,63Fe12O19, respectively. Results of the mean of crystallite size evaluation for respective powder materials showed that the BaxSr1-xFe12O19 (x = 1.0; 0.6; and 0.4) samples have the crystallite size of 22 nm, 25 nm and 34 nm, respectively. We concluded that the cationic distribution of barium atoms was successfully substituted by strontium atoms approaching the nominal stoichiometric composition.

  7. Effect of Buspirone, Fluoxetine and 8-OH-DPAT on Striatal Expression of Bax, Caspase-3 and Bcl-2 Proteins in 6-Hydroxydopamine-Induced Hemi-Parkinsonian Rats

    Hamdollah Sharifi; Alireza Nayebi; Safar Farajnia; Rasool Haddadi

    2015-01-01

    Purpose: The exact pathogenesis of sporadic parkinson’s disease (PD) is still unclear. Numerous evidences suggest involvement of apoptosis in the death of dopaminergic neurons. In this study we investigated the effect of sub-chronic administration of buspirone, fluoxetine and 8-hydroxy-2-[di-n-propylamino]tetralin (8-OH-DPAT) in 6-hydroxydopamine (6-OHDA)-lesioned rats and assayed striatal concentrations of apoptotic (Bax, Caspase3) and anti-apoptotic (Bcl-2) proteins. M...

  8. Der Einfluss der Anästhetika Sevofluran und Propofol auf die Regulation der apoptoseassoziierten Proteine Bax, Bcl-2, Mdm-2 und p53 nach inkompletter zerebraler Hemisphärenischämie bei der Ratte

    Bachl, Monika Maria

    2005-01-01

    Der Einfluss der Anästhetika Sevofluran und Propofol auf apoptoseassoziierte Proteine während zerebraler Ischämie ist bisher nicht erforscht. In der vorliegenden Studie wurden die Effekte dieser Narkotika auf die Regulation der Apoptosefaktoren Bax, Bcl-2, Mdm-2 und p53 bei 36 narkotisierten Sprague-Dawley-Ratten untersucht, bei denen eine inkomplette zerebrale Hemisphärenischämie mit anschließender Reperfusion induziert wurde. Die Apoptosefaktoren wurden mittels Immunfluoreszenz- und Western...

  9. THE EXPRESSIONS OF HIF-1α AND Bax AND THEIR CORRELATION IN SUDDEN CARDIAC DEATH%心脏性猝死心肌组织HIF-1α和Bax表达及其相关性

    刘德衍; 袁磊; 杨彦华; 纪萍; 于建宪; 张七一; 林梅

    2011-01-01

    目的 探讨缺氧诱导因子1 alpha(HIF-1α)和Bax蛋白在心脏性猝死(SCD)者心肌组织中的表达与相关性,并为法医学鉴定SCD提供客观依据.方法 应用免疫组织化学方法,检测25例SCD者心肌组织中HIF-1α和Bax蛋白的表达,并以25例非SCD者心肌组织作为对照.结果 SCD者心肌组织HIF-1α与Bax的表达明显高于对照组(uc=6.083、5.573,P<0.01).在SCD心肌组织中,HIF-1α表达与Bax表达呈正相关(r=0.736,P<0.01).结论 HIF-1α与Bax表达的增高与SCD的发生有一定的相关性,HIF-1α与Bax可望作为诊断SCD较为客观的病理形态学指标.%Objective To investigate the expressions of hypoxia-inducible factor-1 alpha (HIF-1α) and Bcl-2 associated X protein (Bax) and their correlation in myocardium of sudden cardiac death (SCD) and provide an objective evidence for forensic identification.Methods The expressions of HIF-la and Bax in myocardium of 25 SCD were detected using immunohistochemical technique, and 25 of non-sudden cardiac death served as controls.Results The expressions of HIF-1α and Bax in myocardium of the SCD were significantly higher than that of the control (Uc = 6.083,5.573;P<0.01), and the expressions were positively correlated (r=0.736,P<0.01) Conclusion The elevation of the expressions of HIF-1α and Bax is associated with SCD to a certain extent.The HIF-1α and Bax will hopefully become a relatively objective pathomorphologic index in the diagnosis of SCD.

  10. Deregulated expression of A1, Bcl-2, Bcl-xL, and Mcl-1 antiapoptotic proteins and Bid, Bad, and Bax proapoptotic genes in polycythemia vera patients

    Elainy Patricia Lino Gasparotto

    2011-12-01

    Full Text Available Apoptosis deregulation might have a role in the pathophysiology of polycythemia vera (PV. This study evaluated Bcl-2 molecule expression in CD34+ cells and leukocytes in 12 PV patients. Gene expression was investigated by real time PCR using SybrGreen Quantitect kit and protein expression was evaluated by western-blotting. JAK2 V617F mutation was detected according to Baxter et al (2005. CD34+ cells from PV patients presented higher levels of A1 and Mcl-1 expression (median: 22.6 and 5.2, respectively in comparison with controls (0.9 and 0.5, p=0.004 and p=0.020; while Bcl-2 and Bcl-xL expression decreased in PV patients (0.18 and 1.19 compared with controls (1.39 and 2.01, p=0.006 and p=0.020. CD34+ cells in PV patients showed an elevated Bid expression (14.4 in comparison with healthy subjects (1.0; p=0.002. Patients' leukocytes showed an A1 augmentation (7.41, p=0.001 and a reduced expression of Bax (0.19; p=0.040 and Bad (0.2; p=0.030. There was no correlation between JAK2 V617F allele burden and molecular expression. PV patients showed alterations in Bcl-2 members' expression, which may interfere with control of apoptotic machinery and contribute to disease pathogenesis.A desregulação da apoptose parece participar da fisiopatologia da policitemia vera (PV. Este estudo avaliou a expressão das moléculas da família Bcl-2 em células hematopoéticas CD34 + e leucócitos de 12 pacientes com PV. Foram realizados: a quantificação da expressão gênica por PCR em tempo real utilizando kit Sybrgreen Quantitect, avaliação da expressão de proteínas por western-blot e detecção da mutação JAK2 V617F segundo Baxter et al. (2005. Células CD34 + dos pacientes com PV apresentaram maior expressão de A1 e Mcl-1 (mediana: 22,6 e 5,2, respectivamente em comparação com controles (0,9 e 0,5, p = 0,004 e p = 0,020 e expressão de Bcl-2 e Bcl-xL diminuída nestes pacientes (0,18 e 1,19 em relação aos controles (1,39 e 2,01, p = 0,006 e p = 0

  11. Yeast Bax inhibitor, Bxi1p, is an ER-localized protein that links the unfolded protein response and programmed cell death in Saccharomyces cerevisiae.

    James Cebulski

    Full Text Available Bax inhibitor-1 (BI-1 is an anti-apoptotic gene whose expression is upregulated in a wide range of human cancers. Studies in both mammalian and plant cells suggest that the BI-1 protein resides in the endoplasmic reticulum and is involved in the unfolded protein response (UPR that is triggered by ER stress. It is thought to act via a mechanism involving altered calcium dynamics. In this paper, we provide evidence that the Saccharomyces cerevisiae protein encoded by the open reading frame, YNL305C, is a bona fide homolog for BI-1. First, we confirm that yeast cells from two different strain backgrounds lacking YNL305C, which we have renamed BXI1, are more sensitive to heat-shock induced cell death than wildtype controls even though they have indistinguishable growth rates at 30°C. They are also more susceptible both to ethanol-induced and to glucose-induced programmed cell death. Significantly, we show that Bxi1p-GFP colocalizes with the ER localized protein Sec63p-RFP. We have also discovered that Δbxi1 cells are not only more sensitive to drugs that induce ER stress, but also have a decreased unfolded protein response as measured with a UPRE-lacZ reporter. Finally, we have discovered that deleting BXI1 diminishes the calcium signaling response in response to the accumulation of unfolded proteins in the ER as measured by a calcineurin-dependent CDRE-lacZ reporter. In toto, our data suggests that the Bxi1p, like its metazoan homologs, is an ER-localized protein that links the unfolded protein response and programmed cell death.

  12. Silver Nanoparticles Biosynthesized Using Achillea biebersteinii Flower Extract: Apoptosis Induction in MCF-7 Cells via Caspase Activation and Regulation of Bax and Bcl-2 Gene Expression

    Javad Baharara

    2015-02-01

    Full Text Available Silver nanoparticles (Ag-NPs, the most popular nanoparticles, possess unique properties. Achillea biebersteinii is a plant of the Asteraceae family rich in active antitumor components. The aim of this research was the characterization and investigation of the cytotoxic properties of Ag-NPs synthesized using A. biebersteinii flower extract, on a human breast cancer cell line. The Ag-NPs were synthesized after approximately 180 min of reaction at 40 °C, then they were characterized by UV-visible spectroscopy, Fourier transform infrared spectroscopy (FTIR, transmission electron microscopy (TEM and dynamic light scattering (DLS. The anti-apoptosis effect of Ag-NPs on the MCF-7 cell line was investigated by MTT assay, DAPI and acridine orange staining and caspase activity. The transcriptional expression of bax, bcl-2, caspase-3, -8 and -9 were also evaluated by RT-PCR. The TEM images revealed that the Ag-NPs morphology had a different shape. The DLS indicated that the average hydrodynamic diameter of the biosynthesized Ag-NPs was around 12 nm. By UV-visible spectroscopy the strongest absorbance peak was observed at 460 nm. The FTIR results also showed interaction between the plant extract and Ag-NPs due to the similarity in the peak patterns. The EDS results showed that Ag-NPs display an absorption peak at 3 keV, indicating the presence of the element silver. The Ag-NPs caused a dose-dependent decrease in cell viability, fragmentation in nucleic acid, inhibited the proliferation and induction of apoptosis on MCF-7 by suppressing specific cell cycle genes, and simulation programmed cell dead genes. Further investigation is required to establish the potential of this novel and promising approach in cancer therapy.

  13. The influence of sleep deprivation on expression of apoptosis regulatory proteins p53, bcl-2 and bax following rat tongue carcinogenesis induced by 4-nitroquinoline 1-oxide

    Juliana Noguti

    2013-01-01

    Full Text Available Background: The aim of this study was to evaluate whether paradoxical sleep deprivation could affects the mechanisms and pathways essentials for cancer cells in tongue cancer induced by 4-nitroquinole 1-oxide in Wistar rats. Materials and Methods: For this purpose, the animals were distributed into 4 groups of 5 animals each treated with 50 ppm 4 nitroquinoline 1 oxide (4 NQO solution through their drinking water for 4 and 12 weeks. The animals were submitted to paradoxical sleep deprivation (PSD for 72 h using the modified multiple platform method, which consisted of placing 5 mice in a cage (41 × 34 × 16 cm containing 10 circular platforms (3.5 cm in diameter with water 1 cm below the upper surface. The investigations were conducted using immunohistochemistry of p53, Bax and Bcl-2 proteins related to apoptosis and its pathways. Statistical analysis was performed by Kruskal-Wallis non-parametric test followed by the Dunn′s test using SPSS software pack (version 1.0. P value < 0.05 was considered for statistic significance. Results: Although no histopathological abnormalities were induced in the epithelium after 4 weeks of carcinogen exposure in all groups, in 12 weeks were observed pre-neoplasic lesions. Data analysis revealed statistically significant differences ( P < 0.05 in 4 weeks group for p53 and for bcl-2 and for all immunomarkers after 12 weeks of 4NQO administration. Conclusion: Our results reveal that sleep deprivation exerted alterations in proteins associated with proliferation and apoptosis in carcinogenesis.

  14. bax和p53基因共转染对人舌癌细胞增 殖和凋亡的影晌%Effect on Proliferation and Apoptosis of Human Lingual Carc inoma Cells Cotransfected by bax and p53 Genes

    马英红; 杨绍华; 陈俊杰; 高家让; 彭文珍; 王若菡

    2001-01-01

    Objective To investigate th e effects on prolife ration andapoptosis of human cancerous cells cotransfected by bax apoptosis-in ducing gene and p53 tumor suppressor gene. Methods The chi meric gene pSV-CIP-bax -CAT was constructed in which bax gene was flanked upstream by a 217bp fragment (+822~+1093) of the first intron and a 317bp promoter fragment of human a 1(Ⅰ) colla gen gene . Human lingual carcinoma cell line Tca 8113 (LCC) in culture was respe ctively transfected with pSV-CIP-bax-CAT, and cotransfected with pSV-p53-CA T and pSV-CIP-bax-CAT by using the transfection reagent DOSPER. Res ults Immuno- slot blot and ELISA demonstrated that the expression of bax and p53 genes incr eased remarkably in the transfected and cotransfected LCC, compared with the con trols(LCC transfected with pSV-CIP-CAT and LCC).MTT colorimetric assay,TUNE L fluorecence microscopy and flow cytometry showed that foreign bax or p53 gene inhibited the LCC growth and induced apoptosis(23.9% and 26.1% of inhibitory ra te by bax gene and p53 plus bax genes respectively). Conclusion The ectopic expression of the bax gene in the LCC promoted by the cis-act ing elements of human a1(Ⅰ)collagen gene has obviously synergistic effect on th e proliferation and apoptosis of the LCC cotransfected with p53 gene plus bax gene for 48 hours.%目的 探讨诱导凋亡的bax基因和p53抑癌基因共转染对人癌细胞增殖和凋亡的作用。方法 构建pSV-CIP-bax-CAT嵌合基因。在其bax基因上游含人a1(Ⅰ)型胶原基因的第一内含子217bp片段(+822~+1093)和317bp启动子片段。经阳离子转染试剂DOSPER介导,分别用pSV-CIP-bax-CAT转染、pSV-CIP-bax-CAT和pSV-p53-CAT共转染培养的人舌癌Tca8113细胞系(LCC)。结果 免疫狭缝印迹和ELISA检测证实,转染组和共转染组比对照的bax和p53基因表达量明显增加。MTT显色分析、TUNEL荧光显微镜和流式细胞仪监测结果显示,bax基因或p53基因均具抑

  15. Ferroelectric phase transition and soft-mode behavior in BaxSr1−xTiO3: a refined treatment of a quasi-harmonic model

    The composition and temperature dependences of the soft-mode and ferroelectric phase transition in BaxSr1−xTiO3 (BST) are presented and analyzed within the framework of a refined treatment of a quasi-harmonic model (QHM) for quantum particles. The QHM involves generalized simple microscopic quantum particles with anharmonic on-site double-Morse-type potential and harmonic interactions. The theory uses the variational principle scheme at finite temperature in the mean-field approximation while taking into account the predominant mass effect, the cell volume effect as well as ferroelectric distortion. The soft-mode frequency and the Ba concentration (x) dependence of the ferroelectric phase transition temperature are analyzed and show good agreement with experimental results throughout the full x range (0 ⩽ x ⩽ 1). It is found that BST is paraelectric below the critical impurity concentration xc = 0.01 with a softening of the soft mode. It becomes ferroelectric above this critical concentration (x ⩾ xc), and as the impurity concentration increases the predominant displacive soft-mode behavior stiffens in the ferroelectric phase with much less softening in crystals indicating a possible phase change at high x. Around x′c = 0.5, Tc deviates gradually from linear behavior with a rather slight round stagnation which might characterize an equilibrium ‘region’ where several ferroelectric phases coexist. The proposed x dependence of the soft-mode behavior indicates that the ferroelectric phase transition is of first order in BST with x > x′c, and of second order for low Ba concentrations (xc ⩽ x < x′c), throughout the tricritical point at x = x′c. Finally, the quantum dynamics of the ferroelectric phase transition and the mode softening are also addressed and discussed, suggesting a net increase of quantum effects with a decrease of x for x ⩽ 0.1, and a dominant Curie–Weiss law for x > 0.1. (paper)

  16. Development of ferroelectric correlations in the quantum paraelectric and antiferrodistortive regimes in BaxSr1-xTiO3 (x ≤ 0.10)

    The dielectric response ε(T)  of BaxSr1-xTiO3 (x ≤ 0.1) for compositions at and below the critical composition for the ferroelectric transition has been studied. With progressive Ba substitution, the growth of ferroelectric correlations and the weakening of the Antiferrodistortive (AFD) and the quantum paraelectric (QPE) effects have been studied by monitoring the changes in both the in and out of phase parts of the dielectric response. For the compositions close to pure SrTiO3 (x = 0 and x = 0.02), the temperature dependence exhibits a continuous rise in the in-phase part and no ferroelectric peak, consistent with the QPE behavior. With increasing Ba substitution, the low temperature behavior of the in phase part ε′ progressively changes from a continuous rise to exhibit a weak maximum and finally to a well developed cusp. For higher Ba concentrations, the low temperature peak (T ∼ 50K), which corresponds to ferroelectric correlations, becomes increasingly sharper until at the critical composition, x = 0.10, the system shows a single well defined ferroelectric peak. However, the out of phase response of the x = 0.1 composition exhibited a succession of three BaTiO3 type ferroelectric transitions. For x≤0.04, the out of phase part shows evidence of an ordering around T ∼ 100K, which is the expected AFD ordering temperature. The deviations of the ε′(T) data from the Curie-Weiss law have been analyzed within the frame work of two different theoretical models. It was determined that the dielectric behavior for lower concentrations of Ba (up to x ≤ 0.08) was explainable in terms of a model of non-interacting regions which are themselves homogeneously polarized and undergo a second order phase transition. For the phase boundary composition, i.e., x = 0.1, on the other hand, the data are explainable in terms of the Sherrington and Kirkpatrick model which includes the effects of weak correlations between the polar

  17. Cisplatin-induced apoptosis in non-small-cell lung cancer cells is dependent on Bax- and Bak-induction pathway and synergistically activated by BH3-mimetic ABT-263 in p53 wild-type and mutant cells.

    Matsumoto, Masaru; Nakajima, Wataru; Seike, Masahiro; Gemma, Akihiko; Tanaka, Nobuyuki

    2016-04-29

    Cisplatin is a highly effective anticancer drug for treatment of various tumors including non-small-cell lung cancer (NSCLC), and is especially useful in cases nonresponsive to molecular-targeted drugs. Accumulating evidence has shown that cisplatin activates the p53-dependent apoptotic pathway, but it also induces apoptosis in p53-mutated cancer cells. Here we demonstrated that DNA-damage inducible proapoptotic BH3 (Bcl-2 homology region 3)-only Bcl-2 family members, Noxa, Puma, Bim and Bid, are not involved in cisplatin-induced apoptosis in human NSCLC cell lines. In contrast, the expression of proapoptotic multidomain Bcl-2-family members, Bak and Bax, was induced by cisplatin in p53-dependent and -independent manners, respectively. Moreover, in wild-type p53-expressing cells, cisplatin mainly used the Bak-dependent apoptotic pathway, but this apoptotic pathway shifted to the Bax-dependent pathway by loss-of-function of p53. Furthermore, both Bak- and Bax-induced apoptosis was enhanced by the antiapoptotic Bcl-2 family member, Bcl-XL knockdown, but not by Mcl-1 knockdown. From this result, we tested the effect of ABT-263 (Navitoclax), the specific inhibitor of Bcl-2 and Bcl-XL, but not Mcl-1, and found that ABT-263 synergistically enhanced cisplatin-induced apoptosis in NSCLC cells in the presence or absence of p53. These results indicate a novel regulatory system in cisplatin-induced NSCLC cell apoptosis, and a candidate efficient combination chemotherapy method against lung cancers. PMID:26996126

  18. Synergistic Effect of Subtoxic-dose Cisplatin and TRAIL to Mediate Apoptosis by Down-regulating Decoy Receptor 2 and Up-regulating Caspase-8, Caspase-9 and Bax Expression on NCI-H460 and A549 Cells

    Xiaoyan Zhang

    2013-05-01

    Full Text Available Objective(s: Although tumor necrosis factor-related apoptosis-inducing ligand (TRAIL can selectively induce apoptosis in tumor cells, more than half of tumors including non-small cell lung cancer (NSCLC exhibit TRAIL-resistance. The purpose of this study was to determine whether subtoxic-dose cisplatin and TRAIL could synergistically enhance apoptosis on NSCLC cells and investigate its underlying mechanisms. Materials and Methods:NCI-H460 and A549 cells were treated with TRAIL alone, cisplatin alone or combination treatment in this study. The cytotoxicity was evaluated according to Sulforhodamine B assay, and apoptosis was examined using Hoechst 33342 staining and flow cytometry. The mRNA and protein levels of TRAIL receptors and apoptotic proteins including caspase-8, caspase-9, Bcl-2 and Bax were determined by RT-PCR and Western blotting, respectively. Results:Our results showed that NCI-H460 cells were sensitive to TRAIL, whereas A549 cells were resistant. However, subtoxic-dose cisplatin could enhance the both cells to TRAIL-mediated cell proliferation inhibition and apoptosis. The underlying mechanisms might be associated with the down-regulation of DcR2 and up-regulation of Caspase-8, Caspase-9 and Bax. Conclusion:Subtoxic-dose cisplatin could enhance both TRAIL- sensitive and TRAIL- resistant NSCLC cells to TRAIL-mediated apoptosis. These findings motivated further studies to evaluate such a combinatory therapeutic strategy against NSCLC in the animal models.

  19. Protective Effects of Scutellarin on Type II Diabetes Mellitus-Induced Testicular Damages Related to Reactive Oxygen Species/Bcl-2/Bax and Reactive Oxygen Species/Microcirculation/Staving Pathway in Diabetic Rat

    Lingli Long

    2015-01-01

    Full Text Available The goal of our study is to evaluate the effect of Scutellarin on type II diabetes-induced testicular disorder and show the mechanism of Scutellarin’s action. We used streptozotocin and high-fat diet to establish type II diabetic rat model. TUNEL and haematoxylin and eosin staining were used to evaluate the testicular apoptotic cells and morphologic changes. Immunohistochemical staining was used to measure the expression level of vascular endothelial growth factor and blood vessel density in testes. Oxidative stress in testes and epididymis was tested by fluorescence spectrophotometer and ELISA. The expression of Bcl-2/Bax and blood flow rate in testicular vessels were measured by western blot and Doppler. Our results for the first time showed that hyperglycemia induced apoptotic cells and morphologic impairments in testes of rats, while administration of Scutellarin can significantly inhibit these damages. This effect of Scutellarin is controlled by two apoptotic triggers: ROS/Bcl-2/Bax and ROS/microcirculation/starving pathway.

  20. BAX/BCL-2 mRNA and protein expression in human breast MCF-7 cells exposed to drug vehicles-methanol and dimethyl sulfoxide (DMSO for 24 hrs

    Gbenga Anthony Adefolaju

    2015-01-01

    Full Text Available Background: Methanol and DMSO are commonly used as carrier solvents for lipophilic chemicals in in-vitro experiments. However, very little information is available regarding the effects of these solvents on the expression of pro and anti-apoptotic genes and proteins. Materials and Methods: In this study, we examined the cytotoxic effects of methanol and dimethylsulfoxide at 0.5% (final concentrations recommended for in-vitro toxicity assays on human breast cancer MCF-7 cells. We also investigated the effects of these solvents on the mRNA and immunocytochemical expression of apoptotic proteins BAX and BCL-2. Results: The results of neutral red cell viability assay showed that methanol and DMSO concentrations of 0.5% exhibited no cytotoxic effects on MCF-7 cells following a 24 hour exposure. Gene expression and Immunofluorescence results showed that methanol but not DMSO reduced the expression of the BAX pro-apoptotic protein, while both solvents did not alter the expression of the BCL-2 oncoprotein. Conclusion: Our results suggest that while methanol concentrations at 0.5% may be appropriate for in vitro toxicity studies in human breast cancer MCF-7 cells, it could alter the results of gene and protein expression experiments.

  1. Expression of APE1, Bcl-2 and Bax in retinoblastoma and their clinical significance%APE1、Bcl-2及 Bax在视网膜母细胞瘤中的表达及临床意义

    李静; 李德全

    2015-01-01

    目的:分析APE1、Bcl-2及Bax在视网膜母细胞瘤( Rb)中的表达及临床意义。方法选取2011年9月至2013年11月经病理学检查确诊为Rb患者32例及正常视网膜组织16例作为研究对象,免疫组织化学及Western blot分析APE1、Bcl-2及Bax在Rb中及正常视网膜中的表达,比较其在分化及未分化型Rb中的表达。结果 APE1、Bax及Bcl-2在Rb中呈现出高表达,阳性率分别为90.63%、65.63%及68.75%,与正常组比差异均具有统计学意义(χ2=30.13,χ2=12.31,χ2=16.91, P <0.01),与Western blot结果一致;分化组与未分化组中APE1、Bax存在差异显著(χ2=4.99,χ2=7.85, P <0.05),Bcl-2无统计学差异(χ2=0.73, P >0.01)。结论 Rb的发生发展涉及多个基因及生物学过程,分析APE1、bcl-2及bax在Rb中的表达,对Rb的诊断与治疗有重要的参考价值。%Objective To analyze the expression of APE1, Bcl-2 and Bax in retinoblastoma (Rb) and to evaluate their clinical significance.Methods A total of 32 retinoblastoma patients were enrolled for this study from September 2011 to November 2013.Sixteen normal retinal tissues were collected as control.Immunohistochemistry and Western blot were used to evaluate the expression of APE1, Bcl-2 and Bax in retinoblastoma tumor tissues and normal retina.Their expres-sions in differentiated and undifferentiated Rb were also compared.Results APE1, Bax and Bcl-2 were highly expressed in retinoblastoma with positive rates being 90.63%, 65.63% and 68.75%, respectively, and were significantly higher than in normal retina tissues (χ2 =30.13 for APE1,χ2 =12.31 for Bax, andχ2 =16.91 for Bcl-2;P 0.01).Conclusion The development of Rb involves multiple genes and biological processes.Analysis of the expression of APE1, Bcl-2 and Bax in Rb has important clinical value for the diagnosis and treat-ment of Rb.

  2. Expression and significance of Bcl-2 gene and Bax gene in lung cancer%凋亡相关基因bcl-2和bax在肺癌中的表达及意义

    郭雷; 王福昌; 杨五计

    2001-01-01

    To study the relationship between expression of bcl-2 and baxgene and lung cancer.The bcl-2 and bax gene expression in 51 lung cancer tissues and tissues near cancer were deteeted by immunohistochemical SP method.The positive degree of bcl-2 gene expression in lung cancer was higher than that in tissues near cancer.The positive degree of bax gene expression in lung cancer was lower than that in tissues near cancer.Both differences were very significant(P<0.01).There was no correlations between the immunoreactivity of bcl-2 and bax gene and histology,gender,smoking history,clinical subtypes,stage and lymphatic metastasis(P>0.05).It suggests that bcl-2 and bax gene may play an important role in tumorgenesis and tumor development.The detection and determination of bcl-2 and bax gene in preinvasive lesions may contribute to the early diagnosis of lung cancer.Some drugs can down-regulate bcl-2 expression and speed apoptosis so as to improve the sensitivity to chemical therapy and radiation.%为探讨B细胞淋巴瘤/白血病-2(bcl-2)基因及其同源类似物bax基因的表达与肺癌发生的关系,应用免疫组化SP法检测了51例肺癌患者的肺癌标本及其癌旁组织中bcl-2、bax的表达。结果显示,bcl-2在肺癌中染色比在癌旁组织中深,bax在肺癌中染色比在癌旁组织中浅,两者之间均有极显著差异(P<0.01);bcl-2、bax表达与肺癌的组织分型、患者的性别、吸烟史、临床分型、分期及淋巴结转移无关(P>0.05)。提示:①bcl-2、bax在肺癌发生中具有重要作用。②在癌前病变组织中检测并比较bcl-2和bax的表达,有利于肺癌的早期诊断。③可通过药物使bcl-2表达降低,促进凋亡,提高患者对放、化疗的敏感性。

  3. Caractérisation structurale de l'adsorption des isomères para- et meta- du xylène dans la zéolithe de type faujasite BaX Structural Characterization of Para- and Meta- Xylene in Bax Zeolite

    Mellot C.

    2006-11-01

    Full Text Available La séparation du para-xylène des isomères aromatiques en C8 est réalisée industriellement grâce à l'adsorption sur tamis moléculaire zéolithique. Une amélioration des propriétés de séparation des tamis, et en particulier de leur sélectivité, nécessite, entre autres, une bonne connaissance des interactions entre les molécules d'adsorbat et la structure zéolithique. Pour ce faire, nous avons fait appel à deux techniques de caractérisation physico-chimique : la diffraction des neutrons et la spectroscopie infrarouge. Nous avons étudié une zéolithe BaX de type faujasite sur laquelle nous avons adsorbé, à l'état de corps purs, les isomères para- et méta- du xylène. Cette zéolithe est connue industriellement pour ses propriétés sélectives performantes pour le paraxylène. Les analyses ont été réalisées en considérant tout d'abord un faible taux de remplissage, voisin d'une molécule par supercage de zéolithe, et ensuite à saturation où la zéolithe contient sensiblement trois molécules par supercage. La diffraction des neutrons permet, à basse température, de localiser les molécules dans la zéolithe et de préciser leur interaction avec le cation Ba²+. La spectroscopie infrarouge permet une étude des caractéristiques vibrationnelles de l'adsorbat en fonction du taux de recouvrement. Une synthèse des résultats obtenus à l'aide de ces deux méthodes d'investigation nous a permis de dégager un modèle de remplissage des supercages pour les deux isomères considérés. Ainsi, des différences significatives sont mises en évidence. En ce qui concerne le para-xylène, pour un taux de remplissage inférieur à deux molécules par supercage, les molécules de para-xylène viennent se positionner au voisinage des cations en site SII de la supercage. La troisième molécule de para-xylène introduite dans la zéolithe n'est pas en interaction avec un cation Ba²+ et sera localisée dans un nouveau site F

  4. 溴隐亭对大鼠催乳素瘤细胞表达bcl-2、Bax的影响%Effect of bromocriptine on expression of Bax and bcl-2 in rat prolactinoma

    杨雪梅; 徐春; 梁立武; 程海梅

    2012-01-01

    目的 研究溴隐亭对大鼠催乳素(prolactinoma,PRL)瘤表达bcl-2、Bax的影响.方法 (1)用皮下植入17β-雌二醇的方法制备大鼠催乳素瘤模型,同时设立10只大鼠为对照组;(2)将成功诱发出催乳素瘤的大鼠随机分2组,分别给予自来水(安慰剂组)、溴隐亭(溴隐亭组)灌胃,对照组也给予安慰剂灌胃,用药4周后处死动物,垂体称重,测定PRL、Bax、bcl-2的表达水平.结果 (1)17β-雌二醇组大鼠血清PRL水平[(4236.9±416.9) vs (121.2±12.8) ng/ml]和垂体重量[(62.0±5.1) vs (13.8±1.2) mg]均明显高于对照组(P<0.01),证实17β-雌二醇组成功诱发出大鼠泌乳素瘤.(2)溴隐亭组血清PRL水平和垂体重量低于安慰剂组(P<0.01),bcl-2表达水平较安慰剂组明显下降[(1.8±0.5) vs (4.0±0.6),P<0.01],Bax表达水平明显增高[(4.5±0.6) vs ( 1.0±0.3),P<0.01].结论 抑制bcl-2的表达,刺激Bax的表达,从而促进催乳素瘤细胞的凋亡可能是溴隐亭抗催乳素瘤的重要机制之一.%Objective To study the effect of bromocriptine on expression of Bax and bcl -2 in rat prolactinoma. Methods Firstly, to develop prolactinoma rats model. Adult Wistar rats were divided into two groups at random. The rats in control group were subscutaneously implanted with a blank implant . Rats in 17 p - estradiol group were implanted with 17 p - estradiol - containing implants. Secondly, Rats in 17p - estradiol group were divided into two groups at random, i. e. placebo group and bromocriptine group. Water was given to rats in placebo group. Bromocriptine was orally adminstered to rats in bromocriptine group. After 4 weeks of treatment, all the animals were sacrificed. Each pituitary gland was weighed. Serum prolactin(PRL) levels were measured by RIA. Expression level of Bax and bcl -2 in pituitary tissue were measured by Western blotting. Results (1) The weights of pituitary gland and PRL levels in 17p - estradiol group were significantly higher than those

  5. Downregulation of uPAR and cathepsin B induces apoptosis via regulation of Bcl-2 and Bax and inhibition of the PI3K/Akt pathway in gliomas.

    Ramarao Malla

    Full Text Available BACKGROUND: Glioma is the most commonly diagnosed primary brain tumor and is characterized by invasive and infiltrative behavior. uPAR and cathepsin B are known to be overexpressed in high-grade gliomas and are strongly correlated with invasive cancer phenotypes. METHODOLOGY/PRINCIPAL FINDINGS: In the present study, we observed that simultaneous downregulation of uPAR and cathepsin B induces upregulation of some pro-apoptotic genes and suppression of anti-apoptotic genes in human glioma cells. uPAR and cathepsin B (pCU-downregulated cells exhibited decreases in the Bcl-2/Bax ratio and initiated the collapse of mitochondrial membrane potential. We also observed that the broad caspase inhibitor, Z-Asp-2, 6-dichlorobenzoylmethylketone rescued pCU-induced apoptosis in U251 cells but not in 5310 cells. Immunoblot analysis of caspase-9 immunoprecipitates for Apaf-1 showed that uPAR and cathepsin B knockdown activated apoptosome complex formation in U251 cells. Downregulation of uPAR and cathepsin B also retarded nuclear translocation and interfered with DNA binding activity of CREB in both U251 and 5310 cells. Further western blotting analysis demonstrated that downregulation of uPAR and cathepsin B significantly decreased expression of the signaling molecules p-PDGFR-β, p-PI3K and p-Akt. An increase in the number of TUNEL-positive cells, increased Bax expression, and decreased Bcl-2 expression in nude mice brain tumor sections and brain tissue lysates confirm our in vitro results. CONCLUSIONS/SIGNIFICANCE: In conclusion, RNAi-mediated downregulation of uPAR and cathepsin B initiates caspase-dependent mitochondrial apoptosis in U251 cells and caspase-independent mitochondrial apoptosis in 5310 cells. Thus, targeting uPAR and cathepsin B-mediated signaling using siRNA may serve as a novel therapeutic strategy for the treatment of gliomas.

  6. Bax/Bak-dependent, Drp1-independent Targeting of X-linked Inhibitor of Apoptosis Protein (XIAP) into Inner Mitochondrial Compartments Counteracts Smac/DIABLO-dependent Effector Caspase Activation.

    Hamacher-Brady, Anne; Brady, Nathan Ryan

    2015-09-01

    Efficient apoptosis requires Bax/Bak-mediated mitochondrial outer membrane permeabilization (MOMP), which releases death-promoting proteins cytochrome c and Smac to the cytosol, which activate apoptosis and inhibit X-linked inhibitor of apoptosis protein (XIAP) suppression of executioner caspases, respectively. We recently identified that in response to Bcl-2 homology domain 3 (BH3)-only proteins and mitochondrial depolarization, XIAP can permeabilize and enter mitochondria. Consequently, XIAP E3 ligase activity recruits endolysosomes into mitochondria, resulting in Smac degradation. Here, we explored mitochondrial XIAP action within the intrinsic apoptosis signaling pathway. Mechanistically, we demonstrate that mitochondrial XIAP entry requires Bax or Bak and is antagonized by pro-survival Bcl-2 proteins. Moreover, intramitochondrial Smac degradation by XIAP occurs independently of Drp1-regulated cytochrome c release. Importantly, mitochondrial XIAP actions are activated cell-intrinsically by typical apoptosis inducers TNF and staurosporine, and XIAP overexpression reduces the lag time between the administration of an apoptotic stimuli and the onset of mitochondrial permeabilization. To elucidate the role of mitochondrial XIAP action during apoptosis, we integrated our findings within a mathematical model of intrinsic apoptosis signaling. Simulations suggest that moderate increases of XIAP, combined with mitochondrial XIAP preconditioning, would reduce MOMP signaling. To test this scenario, we pre-activated XIAP at mitochondria via mitochondrial depolarization or by artificially targeting XIAP to the intermembrane space. Both approaches resulted in suppression of TNF-mediated caspase activation. Taken together, we propose that XIAP enters mitochondria through a novel mode of mitochondrial permeabilization and through Smac degradation can compete with canonical MOMP to act as an anti-apoptotic tuning mechanism, reducing the mitochondrial contribution to the

  7. Effect of acrylonitrile on the expression of Bcl-2 and Bax in spermatogenic cell of mice%丙烯腈对小鼠生精细胞Bcl-2、Bax蛋白表达的影响

    裴凌云; 马国燕; 金娜; 陈亚; 冯玉娟; 党瑜慧; 薛红丽; 李芝兰

    2012-01-01

    Objective To explore the effect of acrylonitrile exposure on the expression of Bcl-2 and Bax in mice spermatogenic cells. Methods Based on body weight, 250 SPF Kunming adult male mice were randomly divided into five groups; negative control group (normal saline 0. 01ml/g), three AN exposure groups (intraperitoneal injection of 1.25, 2. 50 or 5. 00 mg/kg of AN daily for 5 days, respectively) and positive control group (intraperitoneal injection of cyclophosphamide 40mg/kg) . Mice were killed in the 7th, 14th, 21st, 28th and 35th day after the first exposure by cervical dislocation. Immunohistochemical method ( SABC ) was used to detect the expression of Bcl-2 and Bax protein in spermatogenic cells. Results The average optical density values of Bcl-2 at five time points of the AN 2. 50 mg/kg group and the 21th day point of the AN 1. 25 mg/kg group were significantly lower than the negative control group ( P < 0. 05 ) . Except the 21st day point of the AN 1. 25 mg/kg group, the mean optical density values of Bax in all time points of AN exposure groups were significantly higher than the negative control group (P < 0. 05 ). The decreased expression of Bcl-2 protein was most distinct in AN 2. 50mg/kg group and the positive control group at all time points. The expression of Bax protein was significantly increased in all groups at the 14th day point. Conclusion The expression of Bel-2 protein could be weakened in spermatogenic cells induced by AN, especially in the AN 2. 50 mg/kg8roup; while the expression of Bax was enhanced, and the amplitude of change in the 14th day point was more obvious.%目的 探讨丙烯腈(AN)暴露对小鼠睾丸生精细胞Bcl-2、Bax蛋白表达水平的影响.方法 将250只成年健康SPF级昆明种雄性小鼠,按体重随机分为阴性对照组、3个AN染毒组和阳性对照组,阴性对照组用生理盐水,各染毒组分别以1.25、2.50、5.00mg/kg AN腹腔注射(注射剂量为0.01 ml/g BW),每天1次,连续5天,阳性

  8. Vitis vinifera seeds extract for the modulation of cytosolic factors BAX-α and NF-kB involved in UVB-induced oxidative stress and apoptosis of human skin cells

    DECEAN, HANA; FISCHER-FODOR, EVA; TATOMIR, CORINA; PERDE-SCHREPLER, MARIA; SOMFELEAN, LIDIA; BURZ, CLAUDIA; HODOR, TUDOR; ORASAN, REMUS; VIRAG, PIROSKA

    2016-01-01

    Background and aims The depletion of the ozone layer allows overexposure of the skin to UV radiation, which is prolonged due to the increasing life expectancy, together with inappropriate life habits contribute to the increasing incidence of cutaneous malignancies. Plant extracts with antioxidant capacities are frequently employed as a means to protect skin against ultraviolet (UV) radiations, thus preventing skin cancers. In the present study we assessed a red grape seed extract (GSE) potential capacities to reduce ultraviolet B (UVB) radiation-induced reactive oxygen species (ROS) and subsequent apoptosis in a human keratinocytes cell line (HaCaT). We identified molecules and pathways modulated by the GSE through which this may exert its photoprotective effect. Methods The GSE was standardized according to its polyphenolic content and the most important biologically active compounds, such as epigallocatechin and epicatechin, catechin hydrate, procyanidin B and gallic acid were evidenced by high-performance liquid chromatography. According to the plant extract cytotoxicity on the HaCaT cell line, two concentrations were selected for testing from the non-toxic range: GSE1 (37.5 μgEqGA/ml) and GSE2 (75 μgEqGA/ml). The level of ROS was evaluated with CM-H2DCFDA assay, while apoptosis, Bax-α and NF-kβ p65 proteins with ELISA and confirmed by western-blot. Results Both concentrations of the extract decreased the level of ROS in UVB-irradiated keratinocytes (p<0.001), whereas apoptosis and Bax-α pro-apoptotic protein were only reduced by the higher concentration (GSE2). The NF-kB p65 protein level registered increasing values in time after UVB exposure of the cells, while the tested plant extract re-established its level when its smaller concentration was used (GSE1). Conclusion These results encourage further studies on this extract in order to identify other molecules and pathways through which this extract might exert its beneficial effects and also recommend

  9. Pulsed laser deposition of lead-free (Na0.5Bi0.5)1-xBaxTiO3 ferroelectric thin films with enhanced dielectric properties

    Andrei, A.; Scarisoreanu, N. D.; Birjega, R.; Dinescu, M.; Stanciu, G.; Craciun, F.; Galassi, C.

    2013-08-01

    Ferroelectric lead-free (Na0.5Bi0.5)1-xBaxTiO3 thin films obtained by pulsed laser deposition have been structurally and electrically investigated for compositions, x = 0 and x = 0.06, in and out of the morphotropic phase boundary (MPB). Sodium bismuth titanate Na0.5Bi0.5TiO3 (NBT), pure or in solid solution with other materials (like BaTiO3), is considered to be the best candidate material for lead-free ferroelectric and piezoelectric applications such as actuators and nonvolatile memory devices. Bulk solid solutions with BaTiO3 (BT), (1-x)NBT-xBT (NBT-x%BT) have been investigated widely, also due to a morphotropic phase boundary (MPB) with enhanced dielectric and ferroelectric properties between a rhombohedral and a tetragonal ferroelectric phase, at x = 0.06. Nonetheless, to transpose bulk properties to NBT-BT thin films is a major achievement. XRD technique has been used for structural characterizations of NBT-BT films. Dielectric spectroscopy measurements were performed at room temperature in the frequency range 100 Hz-1 MHz. The best films show pure perovskite phase and good crystalline structure, as a function of specific deposition conditions. Unusual characteristics, especially dielectric constant values higher than those for bulk, have been found for films with specific crystallographic orientations.

  10. Persea declinata (Bl. Kosterm Bark Crude Extract Induces Apoptosis in MCF-7 Cells via G0/G1 Cell Cycle Arrest, Bcl-2/Bax/Bcl-xl Signaling Pathways, and ROS Generation

    Putri Narrima

    2014-01-01

    Full Text Available Persea declinata (Bl. Kosterm is a member of the Lauraceae family, widely distributed in Southeast Asia. It is from the same genus with avocado (Persea americana Mill, which is widely consumed as food and for medicinal purposes. In the present study, we examined the anticancer properties of Persea declinata (Bl. Kosterm bark methanolic crude extract (PDM. PDM exhibited a potent antiproliferative effect in MCF-7 human breast cancer cells, with an IC50 value of 16.68 µg/mL after 48 h of treatment. We observed that PDM caused cell cycle arrest and subsequent apoptosis in MCF-7 cells, as exhibited by increased population at G0/G1 phase, higher lactate dehydrogenase (LDH release, and DNA fragmentation. Mechanistic studies showed that PDM caused significant elevation in ROS production, leading to perturbation of mitochondrial membrane potential, cell permeability, and activation of caspases-3/7. On the other hand, real-time PCR and Western blot analysis showed that PDM treatment increased the expression of the proapoptotic molecule, Bax, but decreased the expression of prosurvival proteins, Bcl-2 and Bcl-xL, in a dose-dependent manner. These findings imply that PDM could inhibit proliferation in MCF-7 cells via cell cycle arrest and apoptosis induction, indicating its potential as a therapeutic agent worthy of further development.

  11. Triphala Extract Suppresses Proliferation and Induces Apoptosis in Human Colon Cancer Stem Cells via Suppressing c-Myc/Cyclin D1 and Elevation of Bax/Bcl-2 Ratio

    Ramakrishna Vadde

    2015-01-01

    Full Text Available Colon cancer is the second leading cause of cancer related deaths in the USA. Cancer stem cells (CSCs have the ability to drive continued expansion of the population of malignant cells. Therefore, strategies that target CSCs could be effective against colon cancer and in reducing the risk of relapse and metastasis. In this study, we evaluated the antiproliferative and proapoptotic effects of triphala, a widely used formulation in Indian traditional medicine, on HCT116 colon cancer cells and human colon cancer stem cells (HCCSCs. The total phenolic content, antioxidant activity, and phytochemical composition (LC-MS-MS of methanol extract of triphala (MET were also measured. We observed that MET contains a variety of phenolics including naringin, quercetin, homoorientin, and isorhamnetin. MET suppressed proliferation independent of p53 status in HCT116 and in HCCSCs. MET also induced p53-independent apoptosis in HCCSCs as indicated by elevated levels of cleaved PARP. Western blotting data suggested that MET suppressed protein levels of c-Myc and cyclin D1, key proteins involved in proliferation, and induced apoptosis through elevation of Bax/Bcl-2 ratio. Furthermore, MET inhibited HCCSCs colony formation, a measure of CSCs self-renewal ability. Anticancer effects of triphala observed in our study warrant future studies to determine its efficacy in vivo.

  12. Persea declinata (Bl.) Kosterm Bark Crude Extract Induces Apoptosis in MCF-7 Cells via G0/G1 Cell Cycle Arrest, Bcl-2/Bax/Bcl-xl Signaling Pathways, and ROS Generation.

    Narrima, Putri; Paydar, Mohammadjavad; Looi, Chung Yeng; Wong, Yi Li; Taha, Hairin; Wong, Won Fen; Ali Mohd, Mustafa; Hadi, A Hamid A

    2014-01-01

    Persea declinata (Bl.) Kosterm is a member of the Lauraceae family, widely distributed in Southeast Asia. It is from the same genus with avocado (Persea americana Mill), which is widely consumed as food and for medicinal purposes. In the present study, we examined the anticancer properties of Persea declinata (Bl.) Kosterm bark methanolic crude extract (PDM). PDM exhibited a potent antiproliferative effect in MCF-7 human breast cancer cells, with an IC50 value of 16.68 µg/mL after 48 h of treatment. We observed that PDM caused cell cycle arrest and subsequent apoptosis in MCF-7 cells, as exhibited by increased population at G0/G1 phase, higher lactate dehydrogenase (LDH) release, and DNA fragmentation. Mechanistic studies showed that PDM caused significant elevation in ROS production, leading to perturbation of mitochondrial membrane potential, cell permeability, and activation of caspases-3/7. On the other hand, real-time PCR and Western blot analysis showed that PDM treatment increased the expression of the proapoptotic molecule, Bax, but decreased the expression of prosurvival proteins, Bcl-2 and Bcl-xL, in a dose-dependent manner. These findings imply that PDM could inhibit proliferation in MCF-7 cells via cell cycle arrest and apoptosis induction, indicating its potential as a therapeutic agent worthy of further development. PMID:24808916

  13. A-site compositional effects in Ga-doped hollandite materials of the form BaxCsyGa2x+yTi8‑2x‑yO16: implications for Cs immobilization in crystalline ceramic waste forms

    Xu, Yun; Wen, Yi; Grote, Rob; Amoroso, Jake; Shuller Nickles, Lindsay; Brinkman, Kyle S.

    2016-06-01

    The hollandite structure is a promising crystalline host for Cs immobilization. A series of Ga-doped hollandite BaxCsyGa2x+yTi8‑2x‑yO16 (x = 0, 0.667, 1.04, 1.33; y = 1.33, 0.667, 0.24, 0) was synthesized through a solid oxide reaction method resulting in a tetragonal hollandite structure (space group I4/m). The lattice parameter associated with the tunnel dimension was found to increases as Cs substitution in the tunnel increased. A direct investigation of cation mobility in tunnels using electrochemical impedance spectroscopy was conducted to evaluate the ability of the hollandite structure to immobilize cations over a wide compositional range. Hollandite with the largest tunnel size and highest aspect ratio grain morphology resulting in rod-like microstructural features exhibited the highest ionic conductivity. The results indicate that grain size and optimized Cs stoichiometry control cation motion and by extension, the propensity for Cs release from hollandite.

  14. A-site compositional effects in Ga-doped hollandite materials of the form BaxCsyGa2x+yTi8−2x−yO16: implications for Cs immobilization in crystalline ceramic waste forms

    Xu, Yun; Wen, Yi; Grote, Rob; Amoroso, Jake; Shuller Nickles, Lindsay; Brinkman, Kyle S.

    2016-01-01

    The hollandite structure is a promising crystalline host for Cs immobilization. A series of Ga-doped hollandite BaxCsyGa2x+yTi8−2x−yO16 (x = 0, 0.667, 1.04, 1.33; y = 1.33, 0.667, 0.24, 0) was synthesized through a solid oxide reaction method resulting in a tetragonal hollandite structure (space group I4/m). The lattice parameter associated with the tunnel dimension was found to increases as Cs substitution in the tunnel increased. A direct investigation of cation mobility in tunnels using electrochemical impedance spectroscopy was conducted to evaluate the ability of the hollandite structure to immobilize cations over a wide compositional range. Hollandite with the largest tunnel size and highest aspect ratio grain morphology resulting in rod-like microstructural features exhibited the highest ionic conductivity. The results indicate that grain size and optimized Cs stoichiometry control cation motion and by extension, the propensity for Cs release from hollandite. PMID:27273791

  15. The structural, magnetic and electrical properties of the hole-doped cobaltites La0.7(Ca1-xBax)0.3CoO3 (x=0.0, 0.5 and 1.0)

    We have studied the structural, magnetic and electrical properties of hole-doped cobaltites with compositions, La0.7(Ca1-xBax)0.3CoO3 (x=0.0, 0.5 and 1.0). Due to the large difference in size between the Ca and Ba ions (size mismatch), these compounds show interesting changes in the magnetic and transport behaviours with an increase in the concentration of Ba. The distortion in the perovskite structure decreases with the substitution of Ba for Ca. Magnetic studies indicate that the anisotropy in their ferromagnetic characters, which contribute to the observations of large thermomagnetic irreversibilities and large coercive fields in these compounds, change significantly with the substitution of Ba for Ca. The unusual magnetic behaviour of the La0.7Ba0.3CoO3 compound, as reflected in its M-H behaviour, has been explained on the basis of a possible coexistence of different magnetic phases. Observations of the low-temperature resistivity minimum, and negative magnetoresistance up to 26% in the low-temperature insulating regions of these polycrystalline compounds, have been ascribed to grain boundary effects. (author)

  16. 甘草黄酮对大强度耐力运动大鼠肾脏组织Ca2+、Mg2+-ATPase及Bax,Bc1-2蛋白表达的影响%Effect of High-Intensity Endurance Exercise on Ca2+,Mg2+-ATPase and Bax, Bcl-2 Protein Expression With Glycyrrhiza Flavonoids in rat Nephridial Tissue

    王东旭; 陈艳艳

    2013-01-01

    Objective To explore Glycyrrhiza Elavonoids on the rat nephridial tissue of Ca2+, Mg2+-ATPase and Bax, Bcl-2 protein expression with high-intensity endurance exercise. Methods The twenty-four healthy male rats were randomly divided into quiet groups, high-intensity exercise group and exercise plus Glycyrrhiza Elavonoids group, After 6 weeks of treadmill training, Using the box of reagent and immunity histochemistry examined the changing of Ca2+, Mg2+-ATPase and Bax, Bcl-2 protein expression on each groups . Results Compared with the quiet groups, the activity of Ca2+, Mg2+-ATPase both had significant droped (P<0.01), and the groups of plus drog had very difference increased than high-intendity exerxise groups (P<0.01); High-intensity endurance exercise group and exercise dosing rats AI apoptosis index increased in varying degrees;high-intensity exercise group (MOD) were very significant difference(P<0.01), exercise plus drug group Bac protein expression (MOD)were very significant difference (P<0.01); Exercise plus drug group Bcl-2 protein expression(MOD) with the high-intersity exercise group had significant difference(P<0.01), High-intensity exercise group and exercise plus drug group Bax/Bcl-2 ratio of distribution is significantly difference degrees of difference(P<0.05,P<0.01).%目的:探讨甘草黄酮对大强度耐力运动大鼠肾脏组织Ca2+、Mg2+-ATPase及Bax、Bcl-2表达的影响。方法:选取SD雄性健康大鼠24只,随机分为安静组、大强度运动组和运动加药组;采用跑台训练6周后取材,应用试剂盒和免疫组织化学法测检测各组大鼠肾脏组织Ca2+、Mg2+-TPase活性及Bax和Bcl-2表达的变化。结果:与安静对照组相比,大强度运动组和运动加药组肾脏组织Ca2+、Mg2+-TPase活性均呈非常显著性下降(P<0.01);其中运动加药组Ca2+、Mg2+-TPase活性均较大强度运动组具有非常显著差异性提高(P<0.01);大强度耐力运动组和运动加

  17. THE EFFECT OF EXERCISE PRECONDITION INDUCED MIR-21 AND BAX GENE EXPRESSIONS ON RATS' CARDIAC MYOCYTE AFTER EXHAUSTIVE EXERCISES%运动预适应对力竭运动后心肌mir-21和bax基因表达的研究

    黄雅雯

    2011-01-01

    [目的]探讨运动预适应(excerise preconditioning,EP)对大鼠一次性力竭运动后心肌mir-21和bax基因表达的影响.[方法]健康雄性SD大鼠36只,随机分为对照组(C组)、一次性力竭运动组(E组)、运动预适应+一次性力竭运动组(EP组).力竭运动后即刻和24 h取各组心肌采用real-time PCR检测各心肌内mir-21和bax基因表达情况.[结果]EP可以显著抑制一次性力竭运动后心肌内mir-21基因的下调表达(P<0.05)和bax基因的上调表达(P<0.05).[结论]EP可以通过抑制mir-21基因下调从而抑制bax基因的上调来发挥保护作用.%[ Objective] To explore the effect of exercise preconditioning induced mir-21 and bax gene expressions on rats' cardiac myocyte after exhaustive exercises. [Methods] 36 healthy male SD rats were randomly divided into control group (C group) , exhaustive exercises (E group) , exercise preconditioning+ exhaustive exercises (EP group) .Used real-time PCR to detect expressions of mir-21 and bax after exhaustive exercises and 24h. [Results] The results of real-time PCR suggested that in EP group, down-regulation of mir-21 and up-regulation of bax cardiac myocyte were obviously inhibited by exhaustive exercises. [Conclusion] EP could inhibit the down-regulation of bax gene through inhibiting up-regulation of mir-21 gene.

  18. Suppression of chondrosarcoma cells by 15-deoxy-Δ12,14-prostaglandin J2 is associated with altered expression of Bax/Bcl-xL and p21

    We previously reported that 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2), the most potent agonist for peroxisome proliferator-activated receptor γ (PPARγ), induces apoptosis of human chondrosarcoma cell line OUMS-27. The current study aimed to explore the mechanism of 15d-PGJ2-induced apoptosis and inhibition of cell proliferation in OUMS-27 cells. The preliminary results of cDNA microarray analysis showed the down-regulation of anti-apoptotic Bcl-xL and up-regulation of pro-apoptotic Bax in the process of 15d-PGJ2-induced apoptosis. These changes were further confirmed at mRNA and protein levels by RT-PCR and Western blot analysis, respectively. Among cyclin-dependent kinase inhibitors, p21 was induced and up-regulated by 15d-PGJ2, but p16 and p27 were not changed, suggesting that the involvement of p21 in inhibition of cell proliferation. Activation of caspase-3 by 15d-PGJ2 was partly, but not completely, blocked by PPARγ antagonist (GW9662) suggesting the 15d-PGJ2 exerted its effect by PPARγ-dependent and -independent pathways. Interestingly, immunohistochemical study on human chondrosarcoma samples revealed that Bcl-xL is frequently expressed by tumor cells. The results of the current study suggest that the potential ability of 15d-PGJ2 in regulation of cell cycle and inhibition of Bcl-xL expression might be beneficial in the development of novel pharmacological agents for chondrosarcoma

  19. All-trans retinoic acid inhibits KIT activity and induces apoptosis in gastrointestinal stromal tumor GIST-T1 cell line by affecting on the expression of survivin and Bax protein

    Taguchi Takahiro

    2010-12-01

    Full Text Available Abstract Background Imatinib, a selective tyrosine kinase inhibitor, has been used as a standard first-line therapy for irresectable and metastasized gastrointestinal stromal tumor (GIST patients. Unfortunately, most patients responding to imatinib will eventually exhibit imatinib-resistance, the cause of which is not fully understood. The serious clinical problem of imatinib-resistance demands alternative therapeutic strategy. This study was conducted to investigate the effect of all-trans retinoic acid (ATRA on GIST cell lines. Methods Cell proliferation was determined by trypan blue dye exclusion test. Western blot analysis was performed to test the expression of activated KIT, its downstream proteins, and apoptosis associated proteins. The cytotoxic interactions of imatinib with ATRA were evaluated using the isobologram of Steel and Peckham. Results and conclusion In this work, for the first time we have demonstrated that ATRA affected on cell proliferation of GIST-T1 and GIST-882 cell line through inhibition of cell growth in a dose dependent manner and induced apoptosis. High dose of ATRA induced morphologic change in GIST-T1 cells, rounded-up cells, and activated the caspase-3 protein. In further examination, we found that the ATRA-induced apoptosis in GIST-T1 cells was accompanied by the down-regulated expression of survivin and up-regulated expression of Bax protein. Moreover, ATRA suppressed the activity of KIT protein in GIST-T1 cells and its downstream signal, AKT activity, but not MAPK activity. We also have demonstrated that combination of ATRA with imatinib showed additive effect by isobologram, suggesting that the combination of ATRA and imatinib may be a novel potential therapeutic option for GIST treatment. Furthermore, the scracht assay result suggested that ATRA was a potential reagent to prevent the invasion or metastasis of GIST cells.

  20. Electric-field-controlled interface strain coupling and non-volatile resistance switching of La1-xBaxMnO3 thin films epitaxially grown on relaxor-based ferroelectric single crystals

    Zheng, Ming; Zhu, Qiu-Xiang; Li, Xue-Yan; Yang, Ming-Min; Wang, Yu; Li, Xiao-Min; Shi, Xun; Luo, Hao-Su; Zheng, Ren-Kui

    2014-09-01

    We have fabricated magnetoelectric heterostructures by growing ferromagnetic La1-xBaxMnO3 (x = 0.2, 0.4) thin films on (001)-, (110)-, and (111)-oriented 0.31Pb(In1/2Nb1/2)O3-0.35Pb(Mg1/3Nb1/2)O3-0.34PbTiO3 (PINT) ferroelectric single-crystal substrates. Upon poling along the [001], [110], or [111] crystal direction, the electric-field-induced non-180° domain switching gives rise to a decrease in the resistance and an enhancement of the metal-to-insulator transition temperature TC of the films. By taking advantage of the 180° ferroelectric domain switching, we identify that such changes in the resistance and TC are caused by domain switching-induced strain but not domain switching-induced accumulation or depletion of charge carriers at the interface. Further, we found that the domain switching-induced strain effects can be efficiently controlled by a magnetic field, mediated by the electronic phase separation. Moreover, we determined the evolution of the strength of the electronic phase separation against temperature and magnetic field by recording the strain-tunability of the resistance [(ΔR/R)strain] under magnetic fields. Additionally, opposing effects of domain switching-induced strain on ferromagnetism above and below 197 K for the La0.8Ba0.2MnO3 film and 150 K for the La0.6Ba0.4MnO3 film, respectively, were observed and explained by the magnetoelastic effect through adjusting the magnetic anisotropy. Finally, using the reversible ferroelastic domain switching of the PINT, we realized non-volatile resistance switching of the films at room temperature, implying potential applications of the magnetoelectric heterostructure in non-volatile memory devices.

  1. Identification of human ferritin, heavy polypeptide 1 (FTH1) and yeast RGI1 (YER067W) as pro-survival sequences that counteract the effects of Bax and copper in Saccharomyces cerevisiae.

    Eid, Rawan; Boucher, Eric; Gharib, Nada; Khoury, Chamel; Arab, Nagla T T; Murray, Alistair; Young, Paul G; Mandato, Craig A; Greenwood, Michael T

    2016-03-01

    Ferritin is a sub-family of iron binding proteins that form multi-subunit nanotype iron storage structures and prevent oxidative stress induced apoptosis. Here we describe the identification and characterization of human ferritin, heavy polypeptide 1 (FTH1) as a suppressor of the pro-apoptotic murine Bax sequence in yeast. In addition we demonstrate that FTH1 is a general pro-survival sequence since it also prevents the cell death inducing effects of copper when heterologously expressed in yeast. Although ferritins are phylogenetically widely distributed and are present in most species of Bacteria, Archaea and Eukarya, ferritin is conspicuously absent in most fungal species including Saccharomyces cerevisiae. An in silico analysis of the yeast proteome lead to the identification of the 161 residue RGI1 (YER067W) encoded protein as a candidate for being a yeast ferritin. In addition to sharing 20% sequence identity with the 183 residue FTH1, RGI1 also has similar pro-survival properties as ferritin when overexpressed in yeast. Analysis of recombinant protein by SDS-PAGE and by electron microscopy revealed the expected formation of higher-order structures for FTH1 that was not observed with Rgi1p. Further analysis revealed that cells overexpressing RGI1 do not show increased resistance to iron toxicity and do not have enhanced capacity to store iron. In contrast, cells lacking RGI1 were found to be hypersensitive to the toxic effects of iron. Overall, our results suggest that Rgi1p is a novel pro-survival protein whose function is not related to ferritin but nevertheless it may have a role in regulating yeast sensitivity to iron stress. PMID:26886577

  2. The effects of the Reinforcing Kidney and Activating Blood Recipe Affecting the Value of bcl-2, bax and caspase-3 in the Tissue of Corpus Cavernosum Smooth Muscle of DMED- Rats%补肾活血合剂对糖尿病阳痿大鼠阴茎平滑肌组织中Bcl-2 Bax和Caspase-3表达的影响

    张国豪; 方再军; 黄青松

    2011-01-01

    目的:观察补肾活血合剂对糖尿病阳痿(DMED)大鼠阴茎平滑肌组织中Bcl-2、Bax、Caspase-3表达的影响.方法:将实验小鼠用2%链脲佐菌素液(STZ)接60mg/kg,腹腔注射建立糖尿病模型,然后在每只糖尿病模型大鼠颈项处注射阿朴吗啡(APOsigma公司)80μg/kg,录相记录阴茎勃起次数.筛选DMED模型,将造成DMED模型随机分为正常组、模型组、中药高、低剂量组、达美康组、达美康+安雄组.除正常组外,其余各组连续给药12周,然后处死测定阴茎平滑肌组织中Bcl-2,Bax,Caspase-3的表达情况.结果:补肾活血合剂对Bc1-2,Bax,Caspase-3有改善作用.结论:补肾活血合剂对糖尿病阳痿大鼠阴茎平滑肌组织中Bcl-2,Bax,Caspase-3有调控效果.%Objective: Observing the effects of reinforcing kidney and activating blood recipe affecting the value of bcl - 2, bax and caspase - 3 in the tissue of corpus cavernosum Smooth muscle of DMED - rats. Methods: An animal model of diabetes was induced by a single intravenous dose of streptozotocin (STZ, 60mg/kg body weight) in Wistar rats. Recording the numbers of penile erection of each rat by camera after subcutaneous inject of apomorphine (80mg/kg).Screening the model of STZ - rats with erectile dysfunction and grouping into 6 groups: control group, model group, Chinese Medicine group, low- dosage group, Diamicron group, Diamicron combined Testosterone Undecanoate group. After administrating 12 weeks, observed the value of bcl -2, bax and caspase-3 in the tissue of corpus cavernosum smooth muscle of DMED - rats. Results: The reinforcing kidney and activating blood recipe can improve the vulue of bcl - 2,bax and caspase -3. Conclusion: The reinforcing kidney and activating blood recipe has regulatory function for bcl -2,bax and caspase - 3 in the tissue of corpus cavernosum smooth muscle of DMED - rats.

  3. Sundew plant, a potential source of anti-inflammatory agents, selectively induces G2/M arrest and apoptosis in MCF-7 cells through upregulation of p53 and Bax/Bcl-2 ratio

    Ghate, NB; Das, A; Chaudhuri, D; Panja, S; Mandal, N

    2016-01-01

    The worldwide cancer incidences are remarkable despite the advancement in cancer drug discovery field, highlighting the need for new therapies focusing on cancer cell and its microenvironment, including inflammation. Several species of Drosera (family: Droseraceae) are used in various traditional as well as homeopathic systems of medicine. Drosera burmannii Vahl. is also enlisted in French Pharmacopoeia in 1965 for the treatment of inflammatory diseases, including chronic bronchitis, asthma and whooping cough. The present study is designed to substantiate the potential of D. burmannii in in vitro anticancer activity and its relation with anti-inflammatory property. In vitro anticancer study revealed that DBME is inhibiting the proliferation of MCF-7 cells without affecting the viability of other malignant and non-malignant cells. DBME induced G2/M phase arrest and apoptosis in MCF-7 cells by suppressing the expression of cyclin A1, cyclin B1 and Cdk-1 and increasing the expression of p53, Bax/Bcl-2 ratio leading to activation of caspases and PARP degradation. Presence of caspase-8 (Z-IETD-fmk) and caspase-9 (Z-LEHD-fmk) inhibitors alone did prevent the apoptosis partially while apoptosis prevention was significantly observed when used in combination, suggesting vital role of caspases in DBME-induced apoptosis in MCF-7 cells. DBME also downregulated LPS-induced increased expression of iNOS, COX-2 and TNF-α along with suppression on intracellular ROS production that confirms the potential of DBME as anti-inflammatory extract. GCMS analysis revealed the presence of four major compounds hexadecanoic acid, tetradecanoic acid, hexadecen-1-ol, trans-9 and 1-tetradecanol along with some other fatty acid derivatives and carotenoids (Beta-doradecin) in DBME. These findings confirmed the anti-inflammatory activity of DBME, which is already listed in French Pharmacopeia in 1965. Here we have additionally reported the anti-breast cancer activity of DBME and its relation to the

  4. Sundew plant, a potential source of anti-inflammatory agents, selectively induces G2/M arrest and apoptosis in MCF-7 cells through upregulation of p53 and Bax/Bcl-2 ratio.

    Ghate, N B; Das, A; Chaudhuri, D; Panja, S; Mandal, N

    2016-01-01

    The worldwide cancer incidences are remarkable despite the advancement in cancer drug discovery field, highlighting the need for new therapies focusing on cancer cell and its microenvironment, including inflammation. Several species of Drosera (family: Droseraceae) are used in various traditional as well as homeopathic systems of medicine. Drosera burmannii Vahl. is also enlisted in French Pharmacopoeia in 1965 for the treatment of inflammatory diseases, including chronic bronchitis, asthma and whooping cough. The present study is designed to substantiate the potential of D. burmannii in in vitro anticancer activity and its relation with anti-inflammatory property. In vitro anticancer study revealed that DBME is inhibiting the proliferation of MCF-7 cells without affecting the viability of other malignant and non-malignant cells. DBME induced G2/M phase arrest and apoptosis in MCF-7 cells by suppressing the expression of cyclin A1, cyclin B1 and Cdk-1 and increasing the expression of p53, Bax/Bcl-2 ratio leading to activation of caspases and PARP degradation. Presence of caspase-8 (Z-IETD-fmk) and caspase-9 (Z-LEHD-fmk) inhibitors alone did prevent the apoptosis partially while apoptosis prevention was significantly observed when used in combination, suggesting vital role of caspases in DBME-induced apoptosis in MCF-7 cells. DBME also downregulated LPS-induced increased expression of iNOS, COX-2 and TNF-α along with suppression on intracellular ROS production that confirms the potential of DBME as anti-inflammatory extract. GCMS analysis revealed the presence of four major compounds hexadecanoic acid, tetradecanoic acid, hexadecen-1-ol, trans-9 and 1-tetradecanol along with some other fatty acid derivatives and carotenoids (Beta-doradecin) in DBME. These findings confirmed the anti-inflammatory activity of DBME, which is already listed in French Pharmacopeia in 1965. Here we have additionally reported the anti-breast cancer activity of DBME and its relation to the

  5. Bax、Bcl-2在局部晚期宫颈癌中的表达及意义%Studies on the Expression and Clinical Significance of Bax and Bcl-2 before neoadjuvant chemotherapy in Local y advanced cervical cancer

    夏红; 易建华; 杨丽; 褚桂芬

    2014-01-01

    Objective To investigate the expression and clinical significance of Bax and Bcl-2 before neo-adjuvant chemotherapy (NACT) in local y advanced cervical cancer (LACC) . Methods 47 patients with stage ⅠB2-ⅡB of LACC treated with neoadjuvant chemotherapy(NACT) between January 2010 and December 2013 in our hospital were retrospectively analyzed. The expressions of Bax and Bcl-2 were determined by means of immunohistochemistry before NACT. Results: The total effective rate of NACT was 72.3%, including CR 14.9%. The Bax and Bcl-2 expression was 55.3% and 36.2% before NACT. In patients who react and react poorly to NACT, the expression of Bax was 64.7% and 38.5% (p0.05). The staining intensity for Bax was related to histopathological grade and lymphatic metastasis (p0.05). The staining intensity for Bcl-2 was not related to the above clinical and pathological factor (P>0.05). Conclusions The NACT of paclitaxel and cisplatin was effective to LACC. The expression of Bax could be related with biological behavior of the tumor and considered as the index to predict efficacy in NACT.%目的:探讨Bax、Bcl-2蛋白在局部晚期宫颈癌(LACC)中的表达及其临床意义。方法应用免疫组化检测47例LACC 组织中化疗前Bax、Bcl-2蛋白的表达情况。结果①47例LACC中,NACT有效率为72.3%,CR 为14.9%。②化疗前,Bax及Bcl-2蛋白阳性表达率分别为:55.3%、36.2%。临床有效及无效病例中,B a x阳性表达率分别为64.7%、38.5%,差异有统计学意义(P0.05)。③肿瘤低分化患者中B a x阳性表达率明显低于高中分化患者,有淋巴结转移者明显低于无淋巴结转移者(P0.05)。Bcl-2表达在各临床病理参数方面均无明显差异(P>0.05)。结论紫杉醇+顺铂静脉化疗对LACC有效,Bax蛋白异常表达与宫颈癌的生物学行为有关,可作为预测和判定NACT疗效的指标。

  6. Investigation of benazepril protection on diabetic myocardium by downregulating the expression of TGF-β1 and Bax%贝那普利下调TGF-β1及Bax表达对糖尿病大鼠心肌的保护作用

    符丽娟; 杨育红; 包翠芬; 刘婉珠

    2011-01-01

    Objective To investigate the effect of benazepril (BZ) on the expression of TGF-β1 and Bax on diabetic myocardium and myocardial hypertrophy. Methods Diabetic animal models were induced by injecting streptozotocin. Model rats were divided into diabetes and BZ groups, control group was also established. BZ was given to treatment group at dose of 10 mg/kg each day by gavage. At 8 weeks, heart mass indexes (HMI) and left ventricular myocardium mass indexes (LVMI) were measured. The expressions of TGF-β1 and Bax in cardiomyocyte were detected and myocardium was studied by electronic microscope. Results Compared with control group, HMI and LVMI were significantly increased in diabetes group ( P < 0. 05 ). The expression of TGF-β1 and Bax were strengthened in diabetic myocardium. Electron microscopic morphometry of heart samples revealed typical diabetic alterations consisting in a focal loss, disorganization of myofihril, mitochondria swelling, collagen proliferation in extracellular matrix. After treatment with BZ for 8 weeks, HMI and LVMI were decreased ( P < 0. 01 ). The expression of TGF-β1 and Bax were weakened and myocardial ultrastructure was better than that of diabetes group. Conclusions TGF-β1 can induce myocardial apoptosis by stimulating the expression of Bax and promote diabetic cardiac hypertrophy, BZ can improve the cardiac function partly due to its effects of downregulating the expression of TGF-β1 and Bax.%目的 探讨贝那普利(BZ)对糖尿病(DM)大鼠心肌转化生长因子β1(TGF-β1)和Bax的表达及心肌肥厚指标的影响.方法 SD大鼠腹腔注射链脲佐菌素(STZ)制备DM模型.实验分为正常对照组(CON)、DM组和BZ治疗组(BZ).治疗组每日灌胃给予贝那普利(10 mg·kg-1·d-1).8 w后测定心脏质量指数(HMI)、左心室质量指数(LVMI),并取左心室心肌免疫组化法检测TGF-β1和 Bax在心肌组织中的表达,透射电镜观察心肌组织超微结构变化.结果 与CON组相比,DM组大

  7. Expression of Cell Apoptosis Protein-Bax in Locally Advanced Cervical Carcinoma after Neoadjuvant Chemotherapy as an Indicator of the Efficiency of the Therapy%局部晚期宫颈癌新辅助化疗后Bax蛋白作为治疗有效性的指标

    游艳琴; 宋磊

    2006-01-01

    探讨Bax蛋白与宫颈癌新辅助化疗疗效之间的关系,用免疫组化方法检测23例局部晚期宫颈癌组织新辅助化疗前后Bax蛋白的表达.结果化疗后2例为临床完全缓解,17例为临床部分缓解,4例患者对新辅助化疗无反应,总有效率为82.61%.NACT前有反应者和无反应者宫颈癌组织的Bax表达无统计学差异(P>0.05).对化疗有反应者NACT后Bax WS值有显著性差异(P<0.05);无反应者NACT后Bax WS值无统计学差异(P>0.05).不同病理类型和临床分期患者NACT前后Bax变化值无显著性差异(P>0.05);不同病理分级患者NACT前后Bax变化值有统计学差异(P<0.05).说明Bax与化疗疗效有显著相关性.NACT前后Bax变化值与临床分期和病理类型无关,而与病理分级有显著相关性.Bax有可能作为判断化疗疗效的指标.

  8. Effect of single prolonged stress on behavior and expression of Bcl-2 and Bax in the rat hippocampus%单一连续应激对大鼠行为学和海马Bcl-2、Bax表达的影响

    徐爱军; 耿菲; 王海涛

    2012-01-01

    Objective To observe the behavior change and the expression of Bcl-2 and Bax in the hippocampus of rats after treated with single-prolonged stress. Method Wistar rats were divided randomly into control group and stress group,and the stress rats were treated with single-prolonged stress. The behavior changes of rats were observed with open-field test, reject grip test and Morries water mace test. Hydrocortisone level in rats blood serum was determined with chemi-luminescence. Immunohistochemistry was used to detect the expressions of Bax and Bal-2 in the hippocampal neurons. . Results Morris water mace test showed that escape latency of control group was 5. 632s ± 1. 065s and stress group was 20. 762s ±3.236s(t = 8.934,P<0.01). Hydrocortisone level of control group was 1.25μg/dl ±0. 12μg/dl and model was 0. 58μg/dl ±0.09μg/dl(t =8.075 ,P <0.01). The ratio of Bcl-2/Bax in control group was 0.904 ±0.109, while the stress group was 1.440 ±0.091(t = 5.458,P <0. 01). Conclusion The neuronal apoptosis in hippocampus may play a role in posttraumatic stress disorder. The increasing of Bcl-2 and Bax protein and the change of Bcl-2/Bax ratio may be the mechanism during single-prolonged stress.%目的 检测单一连续应激(single prolonged stress,SPS)对大鼠行为学和海马神经细胞Bax、Bcl-2表达的影响.方法 将Wistar大鼠分为对照组和应激组.应激组给予SPS应激,采用旷场实验、拒俘反应实验、Morries水迷宫实验等观察大鼠情感行为改变,采用化学发光法测定血清皮质醇浓度,应用免疫组织化学方法检测海马神经元Bcl-2、Bax的表达.结果 对照组水迷宫实验逃避潜伏期为5.632s±1.065s,应激组为20.762s±3.236s(t=9.932,P<0.01);对照组血清皮质醇浓度为1.25 μg/dl ±0.12μg/dl,应激组为0.58μg/dl±0.09μg/dl(t=7.340,P<0.01).应激组海马Bcl-2、Bax表达升高,Bcl-2/Bax上调.结论 SPS应激能使大鼠表现出创伤后应激障碍行

  9. 黄芪多糖对力竭运动大鼠心肌细胞凋亡及BcL-2、Bax蛋白表达的影响%Effects of acute exercise to exhaustion on BcL-2,Bax gene of rat, sheart after supplyment of astragalus polysaccharide

    马兰军; 田振军

    2012-01-01

    为了探讨黄芪多糖(APS)对力竭运动大鼠机体的保护作用,通过建立大强度力竭运动大鼠模型,选取成年健康雄性SD大鼠24只,随机分为安静对照组,运动组和运动加药组,运动组和运动加药组按照训练模型进行为期6周的耐力训练,最后1次训练进行1次力竭运动,随后取心肌组织并进行样本处理.检测大鼠心肌MDA含量,用缺口末端标记法(TUNEL法)及免疫组织化学法检大鼠心肌细胞凋亡指数(AI)及BcL-2、Bax蛋白表达的变化.结果表明:运动组MDA含量、AI和BcL-2、Bax蛋白表达水平均明显高于安静对照组(P<0.05);与运动组比较,运动加药组MDA含量,AI与Bax蛋白表达水平均下降,而BcL-2蛋白表达水平显著增高(P<0.05).黄芪多糖可有效抑制力竭运动大鼠心肌细胞凋亡,此作用可能与其减轻氧化应激、上调BcL-2和下调Bax蛋白表达水平有关.%To study the supplementary astragalus polysaccharide (APS) to exhaustion movement rats myocardial cell apoptosis, observe the influence of myocardial cell apop-tosis BcL-2, the regulation of gene Bax changes, explore astragalus polysaccharide (APS) to exhaustion movement rats airframe of protection. By establishing high intensity exhaustion movement model of rats, the selection of adult health male SD rats 24 only randomly divided into quiet in the exercise group and control group, sports dosing group, the exercise group and exercise dosing groups according to training model six-week endurance training, last training once exhaustion movement, exhaustion after motion myocardial organize and sample processing. Detection rats myocardial MDA content, using gap end labeling (TUNEL law) and the method of immunohis to chemistry methods were used by rats myocardial cell apoptosis I,ndex (AI) and BcL-2, Bax protein expression changes. In the exercise group MDA content, AI and BcL-2, Bax protein expression levels were significantly higher in quiet in control group (P

  10. 灯盏花注射液抗新生鼠缺氧缺血性脑损伤的作用及对Bcl-2、Bax蛋白表达的影响%The anti-injury effect of Breviscapine injection on the hypoxic ischemic brain damage of neonatal rats and the expression of Bcl-2 and Bax

    张明艳; 范淑娟; 李利平; 武变瑛; 王宇

    2011-01-01

    目的:观察灯盏花注射液对新生大鼠缺氧缺血脑损伤(HIBD)的保护作用及对Bcl-2、Bax蛋白表达的影响.方法:采用新生7日龄SD大鼠缺氧缺血性脑损伤模型,设立假手术组、缺氧缺血脑损伤模型组、灯盏花注射液治疗大、中、小剂量组、无菌注射用水对照组.采用硫堇染色、免疫组织化学染色的方法测定各组大鼠海马CA1区神经元密度、组织学分级及凋亡相关基因Bcl-2、Bax蛋白表达情况,并计数各时间点Bcl-2、Bax免疫阳性细胞数目及测定积分光密度值.结果:假手术组,大鼠海马CA1区无锥体细胞缺失,未见明显免疫阳性细胞.与假手术组比较,缺氧缺血脑损伤模型组、无菌注射用水对照组Bcl-2、Bax蛋白表达均于3 d时达到高峰(与其余各时间点比较差别有显著意义P<0.05),神经元密度明显降低,组织学分级显著增高,积分光密度值增加.灯盏花治疗组,与无菌注射用水对照组比较,Bcl-2蛋白表达进一步增加,积分光密度值增加;而Bax蛋白表达则减少,积分光密度值降低;神经元密度显著高于对照组,组织学分级明显降低.结论:灯盏花注射液可能是通过上调Bcl-2表达,抑制Bax表达,减轻缺氧缺血引起的神经元凋亡及迟发性神经元死亡.%Objective: To observe the protective effect of Breviscapine injection on the hypoxic ischemic brain damage of neonatal rats, and the expression of Bcl-2 and Bax. Methods: In this experiment 7-day-old newborn rat with hypoxic-ischemic brain damage model was used and divided into four groups: sham group, model group, control with sterile water for injection group and Breviscapine injection group. Breviscapine injection group was divided into large, medium, and small doses. Used thionin staining and immunohistochemical staining to assay the neuronal density, histological grade, and the expresssion of Bcl-2 and Bax protein in the CA1 hippocampus of each group , the number of

  11. 新生鼠坏死性小肠结肠炎肠上皮细胞Bax表达与细胞凋亡的初步研究%Expression of Bax and apoptosis in enterocytes of neonatal rats with necrotizing enterocolitis

    张勇; 蒋迎佳; 廖志; 叶飘; 郝静梅; 余绍兰; 胡语航

    2014-01-01

    目的 探讨新生鼠坏死性小肠结肠炎(necrotizing enterocolitis,NEC)时肠上皮细胞Bax表达与凋亡相关性.方法 出生当日48只清洁级SD新生鼠采用奇、偶数方法随机分为对照组和NEC模型组,每组24只.对照组由母鼠喂养.模型组与母鼠分开,通过代乳品人工喂养+缺氧-复氧-冷刺激建立新生鼠NEC动物模型.分别于实验开始后24h、48 h、72 h留取大鼠回盲部近段回肠组织行免疫组织化学法检查肠上皮细胞Bax表达,原位末端标记法检测肠上皮细胞凋亡率.结果 与对照组[(666.55±15.77)IOD; (4.73±0.04)%]相比,NEC模型组新生鼠肠上皮细胞Bax表达和细胞凋亡率在实验开始后24 h开始升高[(1 005.06±11.96) IOD; (15.04 ±0.24)%](P<0.01),72 h后达到高峰[(3 340.66±68.72) IOD; (35.65±0.61)%](P<0.01).NEC模型组新生鼠肠上皮细胞凋亡率与肠上皮细胞Bax表达量呈正相关(r=0.94,P<0.01).结论 新生鼠NEC肠上皮细胞凋亡与Bax表达有明确相关性.新生鼠NEC肠上皮细胞可能通过高表达Bax导致肠上皮细胞凋亡.%Objective To investigate the correlation of Bax and apoptosis in enterocytes of neonatal rats with necrotizing enterocolitis (NEC).Methods Forty-eight neonatal rats (1 day) were randomly divided into control group (n =24) and NEC model group (n =24) by use of odd and even.The rats in control group were maternal breast-fed.The rats in NEC model group were separated from their mothers.To be given formula feeding,cold exposure after hypoxic-reoxygenation treatment.The intestinal tissue located at the boundary of ileum and caecum of two groups were gained on the 24 h,48 h and 72 h with which that all rats were sacrificed by cutting neck.Section of intestinal tissue were stained with immunohistochemistry to detect the expression of Bax and were stained with TdT mediated dUTP nick end labeling(TUNEL) to evaluate the apoptosis in each group.Results The integrate optical density (IOD) value of

  12. 中度低温体外循环后大鼠海马bcl-2和bax的表达与神经元凋亡%Hippocampal bcl-2 and bax expressions and neuronal apoptosis after moderate hypothermic cardiopulmonary extracorporeal circulation in rats

    张挺杰

    2005-01-01

    BACKGROUND: Hippocampus injury is wildly believed to involve in neurocognitive dysfunction; the establishment of a rat model of cardiopulmonary bypass(CPB) allows us to investigate the mechanism of CPB-related hippocampus injury.OBJECTIVE: To investigate the effects of moderate hypothermic CPB with a hemodilution on hippocampal bcl-2 and bax gene expression and neuronal apoptosis in rats.DESIGN: A randomized group division study based on the experimental animals.SETTING: Department of anesthesiology in a university hospital.MATERIALS: Thirty Sprague-Dawley (SD) rats were randomly divided into two groups, CPB group and sham-CPB group with 15 rats in each group.METHODS: Total 15 rats of CPB group were subjected to 60-minute moderate hypothermic nonpulsatile CPB using a peristaltic pump and a membrane oxygenator. The CPB circuit was primed with approximately 20 mL 1:1crystaloid-colloid liquid, while another 15 rats of sham-CPB group underwent identical anesthetic and surgical procedures(including cannulation) except CPB itself. At 1 hour post-CPB, six rats in each group were decapitated, and hippocampi were removed, homogenized, and processed for apoptotic gene ( bcl-2 and bax) mRNAs detection. Reverse transcriptase polymerase chain reaction(RT-PCR) is used to detect expression of mRNA by comparing the PCR product of bcl-2 or bax to those of β-actin housekeeping gene. Immunohistochemistry is used to detect bcl-2 and bax protein expressions and terminal deoxynucleiotidyl transferase-mediated dUTP-biotion nick end labeling(TUNEL) staining method was used to detect neuronal apoptosis at 6 hours post-CPB ( n = 6 in each group) . The protein expression was quantitated as percentage of the positively stained area in the total stained. In addition, hippocampal neuronal ultrastructures were studied by electron microscopy at 6 hours post-CPB( n = 3 in each group).ronal apoptosis and ultrastructure changes between the two groups.RESULTS: At 1 hour post-CPB, the expressions of

  13. 氯胺酮对兔膝骨性关节炎软骨细胞Bcl-2及Bax表达的影响%Influence of Ketamine (KET,Ketamine) of intra-articular injection of rabbit chondrocytes and Bax expression

    王雯; 王林; 陆巍

    2015-01-01

    Objective To explore the different concentrations of after the Bcl‐2 ,and the influence of Bax expres‐sion .Method The rest of rabbits with left knee immobilized in extension position to establish osteoarthritis models , then ,divided the models into model group ,ketamine60 group ,ketamine100 group and ketamine200 group in ran‐dom .They were received 0 .4 ml physiologic saline ,KET of 60μmol/L ,KET of 100μmol/L ,KET of 200μmol/L intra‐articular respectively ,two times a week for 4 weeks ,and sacrificed one week after last injection .To return af‐ter knee joint cartilage specimens corresponding processing ,using immunohistochemical method to observe the Bcl‐2 in the articular cartilage and the expression of bax .Result Compared with model group ,the injection of ketamine and promote cartilage cells apoptosis gene Bcl‐2 and Bax genes are changed (P<0 .05) .ketamine200 group of antiapop‐totic effect is the most significant (P<0 .05) .Conclusion Ketamine can suppress the cartilage cell apoptosis induced by osteoarthritis .%目的:观察不同浓度氯胺酮(Ketamine ,KET )家兔关节腔内注射后软骨细胞Bcl‐2及Bax表达的影响。方法通过管型石膏伸直位制动法造模成功后,随机分为模型组、氯胺酮60组、氯胺酮100组、氯胺酮200组,其中模型组、氯胺酮60组、氯胺酮100组、氯胺酮200组分别给予0.4 mL的生理盐水、KET(60μmol/L)、KET(100μmol/L)、KET(200μmol/L)关节腔内注射,每周2次,共进行4周,最后一次注射后1周处死动物。留取膝关节软骨标本作相应处理后,利用免疫组化法观察关节软骨中Bcl‐2及Bax的表达情况。结果与模型组比较,注射氯胺酮的软骨细胞促凋亡基因Bcl‐2及Bax均改变(P<0.05),其中氯胺酮200组的抗凋亡效果最为显著(P<0.05)。结论氯胺酮可以抑制骨关节炎诱导的软骨细胞凋亡。

  14. 吲哚美辛对COPD大鼠TNF-α与Bcl-2和Bax表达的影响%The Influence of Indomethacin on the Expression of TNF-α and Bcl-2 and Bax in Chronic Obstructive Pulmonary Disease Rat Model

    周兰英; 孙圣华; 高健; 宋薇; 杨小仙; 唐文祥

    2012-01-01

    目的:探讨吲哚美辛对慢性阻塞性肺疾病(COPD)模型大鼠趾长伸肌TNF-α及Bcl-2和Bax表达的影响.方法:100只健康Wistar大鼠随机分为模型组和对照组(A组).模型组采用熏香烟和气管内滴注猪胰蛋白酶(PEE)法建立COPD模型大鼠,以低于A组大鼠平均体重的90%判断发生营养不良的标准,将模型组大鼠分为COPD营养正常组(B组)和COPD营养不良组,COPD营养不良组随机化原则分为COPD营养不良生理盐水组(C组)、吲哚美辛1组(D组)、吲哚美辛2组(E组)、吲哚美辛3组(F组).A、B、C三组灌注等量生理盐水,D、E、F组给予不同剂量吲哚美辛(IND)灌胃干预.TUNEL法测定趾长伸肌细胞凋亡率,免疫组织化学法测定Bcl-2、Bax蛋白表达,双抗体夹心法测定血清及趾长伸肌肿瘤坏死因子α(TNF-α)浓度.结果:吲哚美辛干预后,E组趾长伸肌细胞凋亡率、Bcl-2、Bax的蛋白表达、趾长伸肌匀浆和吲哚美辛干预后血清TNF-α浓度低于C、D、F组,但高于A、B组,P<0.05;E组趾长伸肌Bcl-2蛋白表达高于C、D、F组,低于A、B组,P<0.05,结论:Bcl-2、Bax及TNF-α参与COPD模型大鼠趾长伸肌细胞凋亡,适当剂量的吲哚美辛可改善COPD营养不良模型骨大鼠趾长伸肌的凋亡.%Objective: To investigate the effect of indomethacin on the expression of Bcl-2 , TNF-a and Bax in extensor digito-rum longus in the COPD rafs models. Methods: 100 healthy Wistar rats were randomly divided into model group and control group (group A). The model group rat was performed with intratracheal instillation of porcine pancreatic elastase (PEE) and exposed to cigarette smoke to establish COPD models. Malnutrition was defined if the body weight of the rats in the model group was lower than 90% of the mean body weight of the control group. All model rats were divided into non-malnutntion COPD group (group B) and malnutrition COPD group, and the malnutrition COPD group rats were randomly divided

  15. Influence of Zedoray Turmeric Oil on Expression of Bcl- 2/Bax in Human Breast Cancer MCF-7 Cells%莪术油对人乳腺癌MCF-7细胞凋亡及Bcl-2、Bax表达的影响

    杨丽华; 姜杰

    2010-01-01

    目的:观察中药提取物莪术油(zedoray turmeric oil)对人乳腺癌(human breast cancer)细胞系MCF-7的凋亡诱导作用.方法:用不同浓度的莪术油对体外培养的MCF-7细胞进行干预.分别采用MTT、细胞免疫组织化学染色、流式细胞仪检测等方法,观察其对MCF-7细胞增殖的抑制和凋亡诱导作用,并对凋亡相关Bcl-2与Bax蛋白的表达变化进行定性检测.结果:经不同浓度的莪术油处理后的MCF-7细胞,其生长受到明显的抑制,细胞凋亡指数随药物浓度增加而明显增加;莪术油作用后,细胞中Bax蛋白表达明显高于对照组.结论:一定浓度的莪术油能抑制MCF-7细胞增殖,促进其凋亡;其机制可能与调控Bcl-2、Bax 表达有关.

  16. Anisotropic electrical and thermal conductivity in Bi2AE2Co2O8+δ [AE = Ca, Sr1−xBax (x = 0.0, 0.25, 0.5, 0.75, 1.0)] single crystals

    Bi2AE2Co2O8+δ (AE represents alkaline earth), constructed by stacking of rock-salt Bi2AE2O4 and triangle CoO2 layers alternatively along c-axis, is one of promising thermoelectric oxides. The most impressive feature of Bi2AE2Co2O8+δ, as reported previously, is their electrical conductivity mainly lying along CoO2 plane, adjusting Bi2AE2O4 layer simultaneously manipulates both thermal conductivity and electrical conductivity. It in turn optimizes thermoelectric performance of these materials. In this work, we characterize the anisotropic thermal and electrical conductivity along both ab-plane and c-direction of Bi2AE2Co2O8+δ (AE = Ca, Sr, Ba, Sr1−xBax) single crystals. The results substantiate that isovalence replacement in Bi2AE2Co2O8+δ remarkably modifies their electrical property along ab-plane; while their thermal conductivity along ab-plane only has a slightly difference. At the same time, both the electrical conductivity and thermal conductivity along c-axis of these materials also have dramatic changes. Certainly, the electrical resistance along c-axis is too high to be used as thermoelectric applications. These results suggest that adjusting nano-block Bi2AE2O4 layer in Bi2AE2Co2O8+δ cannot modify the thermal conductivity along high electrical conductivity plane (ab-plane here). The evolution of electrical property is discussed by Anderson localization and electron-electron interaction U. And the modification of thermal conductivity along c-axis is attributed to the microstructure difference. This work sheds more light on the manipulation of the thermal and electrical conductivity in the layered thermoelectric materials

  17. Influence of oxidative stress on apoptosis and expression of bax and bcl-2 of enterocytes in burn rats with delayed resuscitation on the plateau%高原地区烧伤后延迟复苏氧化应激对大鼠肠上皮细胞凋亡及bax和bcl-2基因表达的影响

    周文军; 张诚; 刘毅; 刘萍; 马明; 张世范

    2009-01-01

    Objective To explore influence of oxidative stress reaction on apoptosis rate and expres-sion of apoptosis-related genes bax and bcl-2 of enterocytes in severely burned rats with delayed resuscitation on the plateau. Methods One hundred and twenty rats subjected to 30% TBSA full-thickness scald on the back were derided into plateau experimental group (PE, altitude 3840 m) and Lanzhou experimental group (LE, altitude 1517 m). Then LE and PE groups were subdivided into Lanzhou immediate fluid resus-citation group (LIFR, with immediate intraperitoneal injection of isotonic saline after scald, 40 mL/kg), Lanzhou delayed fluid resuscitation group [LDFR, with intraperitoneal injection of isotonic saline at 6 post scald hour (PSH), 40 mL/kg], and plateau immediate fluid resuscitation group (PIFR, with immediate in-traperitoneal injection of isotonic saline after scald, 40 mL/kg), plateau delayed fluid resuscitation group (PDFR, with intraperitoneal injection of isotonic saline at 6 PSH, 40 mL/kg). Another 12 rats were divided into Lanzhou sham scald group (LS) and plateau sham scald group (PS), with 6 rats in each group. Rats in LS and PS groups were sham scalded in a water bath for 15 s without fluid infusion. Rats were sacrificed at 6, 12, 24, 48, 72 PSH for collection of small intestine samples to determine the contents of malonaldehyde (MDA) and total hydrosulfide (TSH). The apoptosis of enterocytes was determined by TUNEL, and the ex-pression of bax and bcl-2 in epithelial cells were observed by immunohistochemical method. Intestinal sample of LS and PS groups were collected to determine the contents of MDA and TSH. Results After being scal-ded, content of MDA in intestinal tissue of rats in LDFR group and PDFR group was respectively greater than that in LIFR group and PIFR group (P<0.05 or P<0.01). Intestinal tissue content of MDA of rats in LDFR group (9.8±4.0 nmol/mg) and PDFR group (10.2±1.3 nmol/mg) was respectively greater than that in LIFR group (9.5±2

  18. ЕКСПРЕСІЯ BCL-2 ТА BAX У ГЛАДКОМ''ЯЗОВИХ ПУХЛИНАХ ТІЛА МАТКИ

    Гончарова, Г.

    2011-01-01

    Некоторые типы лейомиом тела матки сложно отличить от лейомиоcарком, если использовать только рутинные морфологические методы диагностики. Мы исследовали операционный материал 10 пациенток с лейомиомами и 6 пациенток с лейомиосаркомами тела матки и определяли в нем уровень экспрессии bcl-2 и bax в дополнение к стандартной окраске гематоксилин-эозином микропрепаратов опухолевой ткани. Мы определили в большинстве, 8 из 10 лейомиом (80%), позитивную реакцию с bcl-2, в 2 из 10 (20%) была получена...

  19. 8周中等强度低负荷量训练对老龄雌性大鼠骨骼肌Bax和Bcl-2蛋白及SIRT1/SIRT3信号轴基因表达的影响%Effects of 8-week medium intensity low load training on proteins Bax and Bcl-2 and the gene expression of signal axis SIRT1/SIRT3 of skeletal muscle of aged female rats

    李方晖; 肖琳; 覃飞; 刘承宜

    2014-01-01

    In order to observe the effects of 8-week medium intensity low load training on the levels of proteins Bax and Bcl-2 and the gene messenger RNA (mRNA) expression of axis sirtuin 1 (SIRT1)/sirtuin 3 (SIRT3) of gastrocne-mius of aged rats, the authors divided 16 18-month old female SD rats randomly into a control group and an exercise group, each of which contained 8 rats, let the rats in the exercise group do an aerobic exercise on a treadmill for con-secutive 8 weeks, at a speed of 15 km/h (with 60%~75%VO2max), 15 minutes a day, 5 days a week, let the rats in the control group live freely, in 24 hours after rat exercising at the end of week 8, killed the rats, measured gastrocnemius index, measured the levels of proteins Bax and Bcl-2 of gastrocnemius by means of Western blot analysis, measured the mRNA levels of SIRT3, SIRT1, manganese superoxide dismutase (MnSOD), Caspase 3, peroxisome prolifera-tor-activated receptor-γcoactivator-1 (PGC-1α), mitochondrial transcription factor A (TFAM) and nuclear respiratory factor 1 (NRF1) by means of RT-PCR, and revealed the following findings:as for the rats in the exercise group, their gastrocnemius mass and gastrocnemius index increased significantly (P<0.05 and P<0.01 respectively), their protein Bax level decreased significantly (P<0.05), their protein Bcl-2 level and Bcl-2/Bax ratio increased significantly (P<0.05);their mRNA levels of SIRT3, SIRT1, PGC-1α, NRF1, TFAM and MnSOD increased significantly (P<0.05), their mRNA level of Caspase-3 decreased significantly (P<0.05). The said findings indicated the followings:medium intensity low load training could delay the changing of muscle cell apoptosis signal of aged rats; the homeostatic mechanism mediated by axis SIRT1/SIRT3 played an important role in medium intensity low load training increasing the mitochondria refreshing rate and antioxidase level of skeletal muscle of aged rats.%观察8周中等强度低负荷量训练对老龄雌性大鼠腓肠肌Bax和Bcl-2

  20. DNA damage and nucleotide excision repair capacity in healthy individuals

    Slyšková, Jana; Naccarati, Alessio; Poláková, Veronika; Pardini, Barbara; Vodičková, Ludmila; Štětina, R.; Schmuczerová, Jana; Šmerhovský, Z.; Lipská, L.; Vodička, Pavel

    2011-01-01

    Roč. 25, č. 7 (2011), s. 511-517. ISSN 0893-6692 R&D Projects: GA ČR GAP304/10/1286; GA MŠk 7F10069 Grant ostatní: GA MŠk(CZ) GAUK124710 Institutional research plan: CEZ:AV0Z50390512 Keywords : BPDE-induced DNA repair capacity * comet assay * interindividual variability Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.709, year: 2011

  1. Enhancing Survival of Mouse Oocytes Following Chemotherapy or Aging by Targeting Bax and Rad51

    Kujjo, Loro L.; Tiina Laine; Pereira, Ricardo J. G.; Wataru Kagawa; Hitoshi Kurumizaka; Shigeyuki Yokoyama; Perez, Gloria I.

    2010-01-01

    BACKGROUND: Therapeutic approaches to preserve fertility in females undergoing cancer treatments are currently ineffective. This is partly due to limited knowledge of the molecular mechanisms that injured germ cells elicit to repair damage and survive or to abort repair and activate biochemical pathways leading to death. So far, we know that following spontaneously occurring or drug-induced DNA damage, the efficiency of DNA repair is a critical determinant of the cell's fate. The protein enco...

  2. Upregulation of Bax and Bcl-2 following prenatal cocaine exposure induces apoptosis in fetal rat brain

    Xiao, DaLiao; Zhang, Lubo

    2008-01-01

    Cocaine abuse during pregnancy has been associated with numerous adverse perinatal outcomes. Aims: The present study was to determine whether prenatal cocaine exposure induced apoptosis and the possible role of Bcl-2 family genes in the programming cell death in fetal rat brain. Main methods: Pregnant rats were treated with cocaine subcutaneously (30 & 60 mg/kg/day) from day 15 to 21 of gestation. Then the fetal and maternal brains were isolated. Key findings: Cocaine produced a dose-dependen...

  3. Upregulation of Bax and Bcl-2 following prenatal cocaine exposure induces apoptosis in fetal rat brain

    DaLiao Xiao, Lubo Zhang

    2008-01-01

    Cocaine abuse during pregnancy has been associated with numerous adverse perinatal outcomes. Aims: The present study was to determine whether prenatal cocaine exposure induced apoptosis and the possible role of Bcl-2 family genes in the programming cell death in fetal rat brain. Main methods: Pregnant rats were treated with cocaine subcutaneously (30 & 60 mg/kg/day) from day 15 to 21 of gestation. Then the fetal and maternal brains were isolated. Key findings: Cocaine produced a dose-depe...

  4. Farnesyltransferase inhibitor tipifarnib inhibits Rheb prenylation and stabilizes Bax in acute myelogenous leukemia cells

    Ding, Husheng; McDonald, Jennifer S.; Yun, Seongseok; Schneider, Paula A.; Peterson, Kevin L.; Flatten, Karen S.; Loegering, David A.; Ann L Oberg; Riska, Shaun M.; Huang, Shengbing; Sinicrope, Frank A.; Adjei, Alex A.; Judith E Karp; Meng, X. Wei; Kaufmann, Scott H.

    2014-01-01

    Although farnesyltransferase inhibitors have shown promising activity in relapsed lymphoma and sporadic activity in acute myelogenous leukemia, their mechanism of cytotoxicity is incompletely understood, making development of predictive biomarkers difficult. In the present study, we examined the action of tipifarnib in human acute myelogenous leukemia cell lines and clinical samples. In contrast to the Ras/MEK/ERK pathway-mediated Bim upregulation that is responsible for tipifarnib-induced ki...

  5. Involvement of VDAC, Bax and ceramides in the efflux of AIF from mitochondria during curcumin-induced apoptosis.

    Scharstuhl, A.; Mutsaers, H.A.M.; Pennings, S.W.C.; Russel, F.G.M.; Wagener, F.A.D.T.G.

    2009-01-01

    BACKGROUND: We previously identified curcumin as a potent inducer of fibroblast apoptosis, which could be used to treat hypertrophic scar formation. Here we investigated the underlying mechanism of this process. PRINCIPAL FINDINGS: Curcumin-induced apoptosis could not be blocked by caspase-inhibitor

  6. Defect chemistry and transport properties of BaxCe0.85M0.15O3-d

    Wu, J.; Li, L. P.; Espinosa, W. T. P.; Haile, S. M.

    2004-01-01

    The site-incorporation mechanism of M3+ dopants into A2+B4+O3 perovskites controls the overall defect chemistry and thus their transport properties. For charge-balance reasons, incorporation onto the A2+-site would require the creation of negatively charged point defects (such as cation vacancies), whereas incorporation onto the B4+-site is accompanied by the generation of positively charged defects, typically oxygen vacancies. Oxygen-vacancy content, in turn, is relevant to proton-conducting...

  7. Interaction of Insulin-like Growth Factor-binding Protein-3 and BAX in Mitochondria Promotes Male Germ Cell Apoptosis

    Jia, Yue; Lee, Kuk-Wha; Swerdloff, Ronald; Hwang, David; Cobb, Laura J.; Sinha Hikim, Amiya; Lue, Yan He; Cohen, Pinchas; Wang, Christina

    2009-01-01

    Germ cell apoptosis is crucial for spermatogenesis and can be triggered by various stimuli, including intratesticular hormone deprivation. This study proposes a role for insulin-like growth factor binding protein-3 (IGFBP-3) in male germ cell apoptosis. Groups of adult Sprague-Dawley male rats received one of the following treatments for 5 days: (i) daily intratesticular (IT) injections with saline (control); (ii) a single subcutaneous injection of the gonadotropin-releasing hormone antagonis...

  8. Survival factor-induced extracellular signal-regulated kinase phosphorylates BIM, inhibiting its association with BAX and proapoptotic activity

    Harada, Hisashi; Quearry, Bonnie; Ruiz-Vela, Antonio; Korsmeyer, Stanley J.

    2004-01-01

    The “BH3-only” proapoptotic BCL-2 family members initiate the intrinsic apoptotic pathway. A small interfering RNA knockdown of BIM confirms this BH3-only member is important for the cytokine-mediated homeostasis of hematopoietic cells. We show here that the phosphorylation status of BIM controls its proapoptotic activity. IL-3, a hematopoietic survival factor, induces extracellular signal-regulated kinase/mitogen-activated protein kinase-mediated phosphorylation of BIM on three serine sites ...

  9. XRF and SIMS/SNMS analyses of BaxSr1-xTiO3 dielectrics.

    Breuer, U; Krumpen, W; Fitsilis, F

    2003-04-01

    The development of analytical tools and procedures for process control is a prerequisite for the integration of high permittivity and/or ferroelectric materials in CMOS devices. The thickness and composition of perovskite oxide films were determined by wavelength dispersive X-ray fluorescence analysis (XRF) with special emphasis on the ratio of the group-II elements to the Ti content, and a precision of 0.5% was achieved for a typical film thickness of 20-30 nm. Secondary ion mass spectrometry (SIMS) and sputtered neutrals mass spectrometry (SNMS) was used for depth profiling to determine film homogeneity and elemental interdiffusion at hetero-interfaces. Examples are given for Ba(x)Sr(1-x)TiO(3) and SrTiO(x) thin films which were grown in a prototype MOCVD production tool. No interdiffusion was observed for films grown at 600 degrees C on Pt electrodes in contrast to films grown directly on Si. PMID:12707759

  10. Accelerated apoptosis of lymphocytes by augmented induction of Bax in SSI-1 (STAT-induced STAT inhibitor-1) deficient mice

    Naka, Tetsuji; Matsumoto, Tomoshige; Narazaki, Masashi; Fujimoto, Minoru; Morita, Yoshiaki; Ohsawa, Yoshiyuki; SAITO, HIROSHI; Nagasawa, Takashi; Uchiyama, Yasuo; Kishimoto, Tadamitsu

    1998-01-01

    Growth, differentiation, and programmed cell death (apoptosis) are mainly controlled by cytokines. The Janus kinase–signal transducers and activators of transcription (JAK-STAT) signal pathway is an important component of cytokine signaling. We have previously shown that STAT3 induces a molecule designated as SSI-1, which inhibits STAT3 functions. To clarify the physiological roles of SSI-1 in vivo, we generated, here, mice lacking SSI-1. These SSI-1−/− mice displayed growth retardation and d...

  11. Biphasic onset of splenic apoptosis following hemorrhagic shock : critical implications for Bax, Bcl-2, and Mcl-1 proteins

    Hostmann, Arwed; Jasse, Kerstin; Schulze-Tanzil, Gundula; Robinson, Yohan; Oberholzer, Andreas; Ertel, Wolfgang; Tschoeke, Sven K

    2008-01-01

    INTRODUCTION: The innate immune response to trauma hemorrhage involves inflammatory mediators, thus promoting cellular dysfunction as well as cell death in diverse tissues. These effects ultimately bear the risk of post-traumatic complications such as organ dysfunction, multiple organ failure, or adult respiratory distress syndrome. In this study, a murine model of resuscitated hemorrhagic shock (HS) was used to determine the apoptosis in spleen as a marker of cellular injury and reduced immu...

  12. Impact of TGIFLX Expression on the Regulation of BCL2 and BAX in Prostate Cancer Cell Line (LNCaP)

    Raoofian, Reza; Mojarrad, Majid; Heidari, Mansour; Sciences, Tehran, Iran.

    2013-01-01

    Aims: The homeoprotein TGIFLX (transforming growth factor-β-induced factor 2-like, Xlinked), which is essential in male reproduction and development and likely oncogenic when aberrantly expressed in prostate. We have previously shown an aberrant expression of TGIFLX in the majority of human prostate tumors. However, mechanism by which TGIFLX acts in prostate cancer is unknown. The aim of this study was to investigate the effects of overexpression of wild-type TGIFLX (wt-TGIFLX) on LNCaP,...

  13. Probucol Attenuates Cyclophosphamide-induced Oxidative Apoptosis, p53 and Bax Signal Expression in Rat Cardiac Tissues

    Asiri, Yousif A.

    2010-01-01

    Cyclophosphamide (CP) is a widely used drug in cancer chemotherapy and immunosuppression, which could cause toxicity of the normal cells due to its toxic metabolites. Probucol, a cholesterol-lowering drug, acts as potential inhibitor of DNA damage and shows to protect against doxorubicin-induced cardiomyopathy by enhancing the endogenous antioxidant system including glutathione peroxidase, catalase and superoxide dismutase. This study examined the possible protective effects of probucol, a li...

  14. High temperature thermoelectric properties of p-type skutterudites BaxYbyCo4-zFezSb12

    Dong, Y.

    2012-01-01

    Several polycrystalline p-type skutterudites with compositions Ba xYb yCo 4-zFe zSb 12, with varying filler concentrations x and y, and z = 1 to 2, were synthesized by reacting the constituents and subsequent solid state annealing, followed by densification by hot-pressing. Their thermoelectric properties were evaluated from 300 to 820 K. The Yb filling fraction increased with Fe content while the amount of Fe substitution had little influence on the Ba filling fraction. High purity specimens were obtained when the Fe content was low. Bipolar conduction contributed to the thermal conductivity at elevated temperatures. A maximum ZT value of 0.7 was obtained at 750 K for the specimen with the highest Fe content and filling fraction. The potential for thermoelectric applications is also discussed. © 2012 American Institute of Physics.

  15. Staphylococcus aureus Panton-Valentine leukocidin directly targets mitochondria and induces Bax-independent apoptosis of human neutrophils

    Genestier, Anne-Laure; Michallet, Marie-Cécile; Prévost, Gilles; Bellot, Gregory; Chalabreysse, Lara; Peyrol, Simone; Thivolet, Françoise; Etienne, Jerome; Lina, Gérard; Vallette, François M.; Vandenesch, François; Genestier, Laurent

    2005-01-01

    Panton-Valentine leukocidin (PVL) is a pore-forming toxin secreted by Staphylococcus aureus that has recently been associated with necrotizing pneumonia. In the present study, we report that in vitro, PVL induces polymorphonuclear cell death by necrosis or by apoptosis, depending on the PVL concentration. PVL-induced apoptosis was associated with a rapid disruption of mitochondrial homeostasis and activation of caspase-9 and caspase-3, suggesting that PVL-induced apoptosis is preferentially m...

  16. Hypoxia-mediated down-regulation of Bid and Bax in tumors occurs via hypoxia-inducible factor 1-dependent and -independent mechanisms and contributes to drug resistance

    Erler, Janine Terra; Cawthorne, Christopher J; Williams, Kaye J;

    2004-01-01

    Solid tumors with disorganized, insufficient blood supply contain hypoxic cells that are resistant to radiotherapy and chemotherapy. Drug resistance, an obstacle to curative treatment of solid tumors, can occur via suppression of apoptosis, a process controlled by pro- and antiapoptotic members o...

  17. Recombinant human erythropoietin suppresses endothelial cell apoptosis and reduces the ratio of Bax to Bcl-2 proteins in the aortas of apolipoprotein E-deficient mice

    Warren, Jeffrey S.; Zhao, Ying; Yung, Raymond; Desai, Anjali

    2011-01-01

    Recent clinical trials have raised concern that therapy with recombinant human erythropoietin (EPO) may increase cardiovascular disease risk, event rate, and mortality. Endothelial cell (EC) apoptosis has been implicated in both atherogenesis as well as in the destabilization and rupture of atheromatous plaques.

  18. Multiple Doses of Erythropoietin Impair Liver Regeneration by Increasing TNF-α, the Bax to Bcl-xL Ratio and Apoptotic Cell Death

    Katja Klemm; Christian Eipel; Daniel Cantré; Kerstin Abshagen; Menger, Michael D.; Brigitte Vollmar

    2008-01-01

    BACKGROUND: Liver resection and the use of small-for-size grafts are restricted by the necessity to provide a sufficient amount of functional liver mass. Only few promising strategies to maximize liver regeneration are available. Apart from its erythropoiesis-stimulating effect, erythropoietin (EPO) has meanwhile been recognized as mitogenic, tissue-protective, and anti-apoptotic pleiotropic cytokine. Thus, EPO may support regeneration of hepatic tissue. METHODOLOGY: Rats undergoing 68% hepat...

  19. Runx3 Expression Inhibits Proliferation and Distinctly Alters mRNA Expression of Bax in AGS and A549 Cancer Cells

    Torshabi, Maryam; Faramarzi, Mohammad Ali; Tabatabaei Yazdi, Mojtaba; Ostad, Seyyed Naser; Gharemani, Mohammad Hosein

    2011-01-01

    Runx3, a member of Runt-related transcription factor (Runx) proteins with tumor suppressor effect, is a tissue–restricted and cancer related transcription factor that regulate cell proliferation and growth, as well as differentiation. In the present study, exogenous Run3 was transiently expressed in AGS (human gastric adenocarcinoma), with undetectable Runx3 protein and in A549 (human lung carcinoma) with low levels of endogenous Runx3 protein. The GFP tagged Runx3 was transfected into AGS an...

  20. THE EXPRESSION OF BCL-2, BAX AND CASPASE-3 IN NEURON OF THE SPINAL CORD ANTERIOR HORN AFTER CAUDA EQUINA ACUTE COMPRESSION

    王栋; 王展; 李浩鹏; 贺西京

    2006-01-01

    Cauda equina syndrome(CES)is common inclinic,and acute CES are difficult to recover.So,it s very i mportant to i mprove the treat ment ofCES.The study of nerve was turned fromthe si m-ple nerve shift injury research to the neuron changeafter the axon injury.Peripheral nerve injury cancause their central neuron apoptosis was confir medby experi ment,but rare report was observed withthe motor neuron change after the cauda equina in-jury.The present studies want to set the acute CESani mal model and observe th...

  1. A levedura como modelo para o estudo da regulação de Bax por isoformas da proteína cinase C

    Silva, Rui Filipe Duarte da

    2011-01-01

    Tese de doutoramento em Ciências Programmed cell death (PCD), of which apoptosis is the most common morphological expression, is an orchestrated collapse of the cell resulting from the activation of a widely described cascade of molecular events. Due to the importance of this process in tissue homeostasis and development of multicellular organisms, its deregulation is associated with several disorders, including cancer and neurodegenerative diseases. A crucial event in mamma...

  2. Caspase-3和bax在视网膜母细胞瘤中的表达%Expression of caspase-3 and bax gene protein in retinoblastoma

    孙红; 惠延年; 王立勤; 马吉献

    2003-01-01

    目的: 观察凋亡及凋亡调控基因caspase-3/bax在视网膜母细胞瘤(retinoblastoma, RB)中的表达及与凋亡的相关性. 方法: 收集35例RB标本,对其分别进行caspase-3和bax免疫组织化学染色,观察表达情况及染色强度. 结果: Caspase-3及bax在未分化型(n=15)分别有较好的表达(11/12例),caspase-3及bax在分化型(n=20)中也有较好的表达(17/18例). 正常视网膜组织中无caspase-3及bax的表达. 结论: 凋亡在RB中是存在的,caspase-3及bax在RB的发生发展中起重要作用.

  3. Effect of PAMd on proteins expression of Bax and Bcl-2 of nerve cells in the brain tissue of ischemia-reperfusion mice

    GUOLianjun; LVQing; QULing

    2004-01-01

    AIM : To study the effects of PAMd ( Phenolic alkaloids from Menispermum dauricum on Brain ischemia and ischemic reperfusion injury in mice. METHODS : Bilateral carotid arteries of mice were ligated and 0. 3ml blood was letted from post-eyeball venous jungle, one hour later the carotid arteries were loosed. After cerebral ischemia-reperfusion, mice

  4. Spin-orbit Coupling Induced Magnetism in the d-density Wave Phase of La2-xBaxCuO4 Superconductors

    Wu, Congjun; /Stanford U., Phys. Dept. /Santa Barbara, KITP; Zaanen, Jan; Zhang, Shou-Cheng; /Stanford U., Phys. Dept.

    2010-01-15

    We study the effects of spin-orbit coupling in the d-density wave (DDW) phase. In the low-temperature orthorhombic phase of La{sub 2-x}Ba{sub x}CuO{sub 4}, we find that spin-orbit coupling induces ferromagnetic moments in the DDW phase, which are polarized along the [110] direction with a considerable magnitude. This effect does not exist in the superconducting phase. On the other hand, if the d-density wave order does not exist at zero field, a magnetic field along the [110] direction always induces such a staggered orbital current. We discuss experimental constraints on the DDW states in light of our theoretical predictions.

  5. Effects of human interleukin 10 gene transfer on the expression of Bcl-2 Bax and apoptosis of hepatocyte in rats with acute hemorrhagic necrotizing pancreatitis

    GU Jun-chao; WANG Yu; ZHANG Zhong-tao; XUE Jian-guo; LI Jian-she; ZHOU Yan-zhong

    2005-01-01

    @@ Acute necrotising pancreatitis is characterized by inflammatory and necrotic events, which follow the initial intra-acinar injury involving enzyme activation, and disruption of the acinar cytoskeleton.1 At present, apoptosis has become a hot topic in many kinds of disease.

  6. Effects of Exercise Pre-Conditioning on Hippocampus Expression of Bcl-2 and Bax Protein and Apoptosis Following Ischemia/Reperfusion Injury in Male Rats

    Nabi Shamsaei; Hamid Rajabi; Nahid Aboutaleb; Farnaz Nikbakht; Pezhman Motamedi; Mehdi Khaksari; Sohaila Erfani

    2015-01-01

    Introduction: Cerebral ischemia/reperfusion leads to loss of vulnerable neurons by apoptosis in specific brain regions specially in the hippocampus. There is some evidence indicating that the neuroprotective effects of physical activity on the brain. Therefore,the main purpose of this study was to investigate the effect of exercise pre-conditioning on apoptosis-related proteins expression in hippocampal CA1 neurons after induction of ischemia. Methods: 21 Male rats weighing 260-300g were ...

  7. Resveratrol Reverses Cadmium Chloride-induced Testicular Damage and Subfertility by Downregulating p53 and Bax and Upregulating Gonadotropins and Bcl-2 gene Expression

    Eleawa, Samy M.; ALKHATEEB, Mahmoud A; ALHASHEM, Fahaid H; BIN-JALIAH, Ismaeel; SAKR, Hussein F; ELREFAEY, Hesham M; ELKARIB, Abbas O; ALESSA, Riyad M; HAIDARA, Mohammad A; Shatoor, Abdullah S.; KHALIL, Mohammad A

    2014-01-01

    This study was performed to investigate the protective and therapeutic effects of resveratrol (RES) against CdCl2-induced toxicity in rat testes. Seven experimental groups of adult male rats were formulated as follows: A) controls+NS, B) control+vehicle (saline solution of hydroxypropyl cyclodextrin), C) RES treated, D) CdCl2+NS, E) CdCl2+vehicle, F) RES followed by CdCl2 and M) CdCl2 followed by RES. At the end of the protocol, serum levels of FSH, LH and testosterone were measured in all gr...

  8. Time Dependent Bladder Apoptosis Induced by Acute Bladder Outlet Obstruction and Subsequent Emptying is Associated with Decreased MnSOD Expression and Bcl-2/Bax Ratio

    Li, Wen Ji; Shin, Mi-Kyung; Oh, Seung-June

    2010-01-01

    Ischemia/reperfusion (I/R) injury-induced oxidative stress plays an important role in the functional impairment of the bladder following acute bladder outlet obstruction (BOO) via induction of apoptosis. The purpose of this study was to investigate the time course of the bladder apoptosis, and apoptosis related molecular changes in the early stage of acute BOO. Twelve-week-old male Sprague Dawley rats were divided into control, acute BOO only (I), and acute BOO plus subsequent emptying (I/R) ...

  9. BAX-mediated cell death affects early germ cell loss and incidence of testicular teratomas in Dnd1Ter/Ter mice

    Cook, Matthew S; Coveney, Douglas; Batchvarov, Iordan; Nadeau, Joseph H.; Capel, Blanche

    2009-01-01

    A homozygous nonsense mutation (Ter) in murine Dnd1 (Dnd1Ter/Ter) results in a significant early loss of primordial germ cells (PGCs) prior to colonization of the gonad in both sexes and all genetic backgrounds tested. The same mutation also leads to testicular teratomas only on the 129Sv/J background. Male mutants on other genetic backgrounds ultimately lose all PGCs with no incidence of teratoma formation. It is not clear how these PGCs are lost or what factors directly control the strain-s...

  10. Nimbolide Sensitizes Human Colon Cancer Cells to TRAIL through Reactive Oxygen Species- and ERK-dependent Up-regulation of Death Receptors, p53, and Bax*

    Gupta, Subash C.; Reuter, Simone; Phromnoi, Kanokkarn; Park, Byoungduck; Hema, Padmanabhan S.; Nair, Mangalam; Aggarwal, Bharat B.

    2010-01-01

    TNF-related apoptosis-inducing ligand (TRAIL) shows promise as a cancer treatment, but acquired tumor resistance to TRAIL is a roadblock. Here we investigated whether nimbolide, a limonoid, could sensitize human colon cancer cells to TRAIL. As indicated by assays that measure esterase activity, sub-G1 fractions, mitochondrial activity, and activation of caspases, nimbolide potentiated the effect of TRAIL. This limonoid also enhanced expression of death receptors (DRs) DR5 and DR4 in cancer ce...

  11. The role of hRev7, the accessory subunit of hPolζ, in translesion synthesis past DNA damage induced by benzo[a]pyrene diol epoxide (BPDE

    Maher Veronica M

    2010-12-01

    Full Text Available Abstract Background DNA polymerase zeta (Polζ is a specialized DNA polymerase that, unlike classical replicative polymerases, is capable of replicating past DNA lesions, i.e. of performing translesion synthesis (TLS. The catalytic subunit of hPolζ, hRev3, has been shown to play a critical role in DNA damage-induced mutagenesis in human cells, but less is known about the role of hRev7, the accessory subunit of hPolζ, in such mutagenesis. To address this question, we recently generated human fibroblasts with very significantly reduced levels of hRev7 protein and demonstrated that hRev7 is required to protect cells from ultraviolet(254 nm (UV radiation-induced cytotoxicity and mutagenesis (McNally et al., DNA Repair 7 (2008 597-604. The goal of the present study was to determine whether hRev7 is similarly involved in the tolerance of DNA damage induced by benzo[a]pyrene diol epoxide (BPDE, the reactive form of the widespread environmental carcinogen benzo[a]pyrene. Methods To determine whether hRev7 also plays a role in protecting human cells from the cytotoxicity and mutagenesis induced by benzo[a]pyrene diol epoxide (BPDE, cell strains with reduced hRev7 were compared to their parental strain and a vector control strain for the effect of BPDE on cell survival, induction of mutations, and the ability to progress through the cell cycle. Results The results show that cell strains with reduced hRev7 are more sensitive to the cytotoxic effect of BPDE than the control strains, and progress through S-phase at a slower rate than the control cells following BPDE treatment, indicating that hRev7, and likely hPolζ, is required for efficient bypass of BPDE-induced DNA lesions. However, neither the frequency nor kinds of mutations induced by BPDE in cells with reduced hRev7 differ significantly from those induced in the control strains, suggesting that hPolζ is not essential for inserting nucleotides opposite BPDE-induced DNA damage. Conclusions Taken

  12. Effect of Jiangru Power on the Expression of Bcl-2 and Bax in Rat Prolactinoma%中药降乳散对大鼠催乳素瘤细胞表达Bcl-2、Bax的影响

    徐春; 邱文娟; 付淑云; 徐立红

    2009-01-01

    体质量,血清催乳素水平均明显低于雌激素组,具有显著差别(P<0.01).Bax在降乳散组的表达量明显高于雌激素组(P<0.01).Bcl-2在降乳散组的表达量明显低于雌激素组(P<0.01).结论 中药降乳散具有促进催乳素瘤细胞凋亡的作用.本实验从分子水平探讨了中药抗催乳素瘤的机制,为中药治疗催乳素瘤提供了理论基础.

  13. Bcl-2、Bax在慢性高眼压大鼠视网膜的表达%The expression of Bcl-2 and Bax in chronic glaucom model rat retina

    强晓鑫; 侯力华; 马雅玲; 付金东; 何利东

    2011-01-01

    目的 探讨Bcl-2、Bax在慢性高眼压大鼠视网膜的表达,以及与慢性高眼压大鼠视网膜损伤的关系.方法 健康成年SD大鼠20只,分为正常组和实验组,每组各10只.实验组烙闭大鼠巩膜上静脉制作慢性高眼压模型,造模2个月后,通过HE染色观察视网膜神经节细胞丢失情况,用免疫组织化学法检测视网膜中Bcl-2、bax的表达.结果 光镜下观察实验组较正常组神经节细胞数量减少(P<0.05).Bcl-2蛋白在正常组及实验组大鼠视网膜上均有阳性表达,位于神经节细胞层及内核层,实验组较正常组Bcl-2表达程度有增高,差异有统计学意义(P<0.05).正常组大鼠视网膜中Bax蛋白在神经节细胞层有弱阳性表达,实验组Bax蛋白表达位于神经节细胞层和内核层,表达程度明显增强,与正常组比较差异有统计学意义(P<0.05).结论 Bcl-2及Bax的表达失衡可能是慢性高眼压视网膜神经节细胞凋亡的原因之一.

  14. Inclusion Complex of Zerumbone with Hydroxypropyl- β -Cyclodextrin Induces Apoptosis in Liver Hepatocellular HepG2 Cells via Caspase 8/BID Cleavage Switch and Modulating Bcl2/Bax Ratio

    Nabilah Muhammad Nadzri; Ahmad Bustamam Abdul; Mohd Aspollah Sukari; Siddig Ibrahim Abdelwahab; Eid, Eltayeb E. M.; Syam Mohan; Behnam Kamalidehghan; Theebaa Anasamy; Kuan Beng Ng; Suvitha Syam; Ismail Adam Arbab; Heshu Sulaiman Rahman; Hapipah Mohd Ali

    2013-01-01

    Zerumbone (ZER) isolated from Zingiber zerumbet was previously encapsulated with hydroxypropyl- β -cyclodextrin (HP β CD) to enhance ZER's solubility in water, thus making it highly tolerable in the human body. The anticancer effects of this new ZER-HP β CD inclusion complex via apoptosis cell death were assessed in this study for the first time in liver hepatocellular cells, HepG2. Apoptosis was ascertained by morphological study, nuclear stain, and sub-G1 cell population accumulation with G...

  15. Inclusion Complex of Zerumbone with Hydroxypropyl-β-Cyclodextrin Induces Apoptosis in Liver Hepatocellular HepG2 Cells via Caspase 8/BID Cleavage Switch and Modulating Bcl2/Bax Ratio

    Nabilah Muhammad Nadzri

    2013-01-01

    Full Text Available Zerumbone (ZER isolated from Zingiber zerumbet was previously encapsulated with hydroxypropyl-β-cyclodextrin (HPβCD to enhance ZER’s solubility in water, thus making it highly tolerable in the human body. The anticancer effects of this new ZER-HPβCD inclusion complex via apoptosis cell death were assessed in this study for the first time in liver hepatocellular cells, HepG2. Apoptosis was ascertained by morphological study, nuclear stain, and sub-G1 cell population accumulation with G2/M arrest. Further investigations showed the release of cytochrome c and loss of mitochondrial membrane potential, proving mitochondrial dysfunction upon the ZER-HPβCD treatment as well as modulating proapoptotic and anti-apototic Bcl-2 family members. A significant increase in caspase 3/7, caspase 9, and caspase 8 was detected with the depletion of BID cleaved by caspase 8. Collectively, these results prove that a highly soluble inclusion complex of ZER-HPβCD could be a promising anticancer agent for the treatment of hepatocellular carcinoma in humans.

  16. Evolution of the crystalline structure in (Bi{sub 0}.5Na{sub 0}.5){sub 1}-xBaxTiO{sub 3} thin films around the Morpho tropic Phase Boundary

    Perez-Mezcua, D.; Calzada, M. L.; Bretos, I.; Ricote, J.; Chateigner, D.; Escobar-Galindo, R.; Jimenez, R.; Sirera, R.

    2014-02-01

    (Bi{sub 0}.5Na{sub 0}.5){sub 1}-xBa{sub x}TiO{sub 3} (BNBT), which exhibits compositions for the morphotropic phase boundary (MPB) where exist an intimate coexistence of the rhombohedral and tetragonal structures, is being considered as promising lead-free alternative to the well known Pb(Zr{sub x},Ti{sub 1}-x)O{sub 3} (PZT). In this work, BNBT thin films were fabricated by chemical solution deposition (CSD) with a wide range of compositions (x{approx}{approx}0.050-0.150) onto Pt/TiO{sub 2}/SiO{sub 2}/(100)Si substrates. Structural studies by X-ray diffraction ({lambda}Cu{approx}1.5406 A) using a four-circle goniometer were carried out to determine the crystalline structure of the films. Rietveld analysis of the experimental X-ray patterns showed different volume fractions of the rhombohedral and tetragonal phases as a function of the Ba{sup 2}+ content and the coexistence of both phases, characteristic of a MPB region, for x{approx}­0.055-0.080. Finally, Rutherford backscattering experiments (RBS) were performed to determine the compositional profile of the films. This study revealed a homogenous composition of the BNBT films with abrupt film/substrate interfaces. (Author)

  17. Properties and performance of BaxSr1−xCo0.8Fe0.2O3−δ materials for oxygen transport membranes

    Vente, Jaap F.; McIntosh, Steven; Haije, Wim G.; Bouwmeester, Henny J.M.

    2006-01-01

    The present paper discusses the oxygen transport properties, oxygen stoichiometry, phase stability, and chemical and mechanical stability of the perovskites $${\\text{Ba}}_{{0.5}} {\\text{Sr}}_{{0.5}} {\\text{Co}}_{{0.8}} {\\text{Fe}}_{{0.2}} {\\text{O}}_{{3 - \\delta }} $$ (BSCF) and $${\\text{SrCo}}_{{0.

  18. 视网膜母细胞瘤Bcl-2和Bax基因蛋白质表达%Expression of Bcl-2 and Bax gene protein in retinoblastoma

    张小猛; 庞利民; 张晓光

    2000-01-01

    目的:研究凋亡及凋亡调控基因Bcl-2/Bax和视网膜母细胞瘤(Retinoblastoma,RB)的发生、发展及退化的关系.方法:收集36例RB标本,对其分别进行Bcl-2和Bax基因的蛋白质免疫组织化学染色.对其表达情况和染色强度进行观察.结果:①Bcl-2在分化型RB中表达比较好;②Bax在未分化型和分化型中表达都比较好.结论:①分化型和未分化型RB中都有Bcl-2/Bax基因蛋白表达;②随RB恶性度的增加,Bcl-2的表达逐渐减弱;Bax的表达无明显改变.③分化型RB受Bcl-2和Bax基因共同控制;未分化型RB受Bax基因调控,Bcl-2基因发挥很少的作用.

  19. Non-secreted clusterin isoforms are translated in rare amounts from distinct human mRNA variants and do not affect Bax-mediated apoptosis or the NF-κB signaling pathway.

    Hans Prochnow

    Full Text Available Clusterin, also known as apolipoprotein J, is expressed from a variety of tissues and implicated in pathological disorders such as neurodegenerative diseases, ischemia and cancer. In contrast to secretory clusterin (sCLU, which acts as an extracellular chaperone, the synthesis, subcellular localization and function(s of intracellular CLU isoforms is currently a matter of intense discussion. By investigating human CLU mRNAs we here unravel mechanisms leading to the synthesis of distinct CLU protein isoforms and analyze their subcellular localization and their impact on apoptosis and on NF-κB-activity. Quantitative PCR-analyses revealed the expression of four different stress-inducible CLU mRNA variants in non-cancer and cancer cell lines. In all cell lines variant 1 represents the most abundant mRNA, whereas all other variants collectively account for no more than 0.34% of total CLU mRNA, even under stressed conditions. Overexpression of CLU cDNAs combined with in vitro mutagenesis revealed distinct translational start sites including a so far uncharacterized non-canonical CUG start codon. We show that all exon 2-containing mRNAs encode sCLU and at least three non-glycosylated intracellular isoforms, CLU1‑449, CLU21‑449 and CLU34‑449, which all reside in the cytosol of unstressed and stressed HEK‑293 cells. The latter is the only form expressed from an alternatively spliced mRNA variant lacking exon 2. Functional analysis revealed that none of these cytosolic CLU forms modulate caspase-mediated intrinsic apoptosis or significantly affects TNF-α-induced NF-κB-activity. Therefore our data challenge some of the current ideas regarding the physiological functions of CLU isoforms in pathologies.

  20. Protective Effects of Scutellarin on Type II Diabetes Mellitus-Induced Testicular Damages Related to Reactive Oxygen Species/Bcl-2/Bax and Reactive Oxygen Species/Microcirculation/Staving Pathway in Diabetic Rat

    Long, Lingli; Wang, Jingnan; Lu, Xiaofang; Xu, Yuxia; Zheng, Shuhui; Luo, Canqiao; Li, Yubin

    2015-01-01

    The goal of our study is to evaluate the effect of Scutellarin on type II diabetes-induced testicular disorder and show the mechanism of Scutellarin's action. We used streptozotocin and high-fat diet to establish type II diabetic rat model. TUNEL and haematoxylin and eosin staining were used to evaluate the testicular apoptotic cells and morphologic changes. Immunohistochemical staining was used to measure the expression level of vascular endothelial growth factor and blood vessel density in ...

  1. Protective Effects of Scutellarin on Type II Diabetes Mellitus-Induced Testicular Damages Related to Reactive Oxygen Species/Bcl-2/Bax and Reactive Oxygen Species/Microcirculation/Staving Pathway in Diabetic Rat

    Lingli Long; Jingnan Wang; Xiaofang Lu; Yuxia Xu; Shuhui Zheng; Canqiao Luo; Yubin Li

    2015-01-01

    The goal of our study is to evaluate the effect of Scutellarin on type II diabetes-induced testicular disorder and show the mechanism of Scutellarin’s action. We used streptozotocin and high-fat diet to establish type II diabetic rat model. TUNEL and haematoxylin and eosin staining were used to evaluate the testicular apoptotic cells and morphologic changes. Immunohistochemical staining was used to measure the expression level of vascular endothelial growth factor and blood vessel density in ...

  2. Persea declinata (Bl.) Kosterm Bark Crude Extract Induces Apoptosis in MCF-7 Cells via G0/G1 Cell Cycle Arrest, Bcl-2/Bax/Bcl-xl Signaling Pathways, and ROS Generation

    2014-01-01

    Persea declinata (Bl.) Kosterm is a member of the Lauraceae family, widely distributed in Southeast Asia. It is from the same genus with avocado (Persea americana Mill), which is widely consumed as food and for medicinal purposes. In the present study, we examined the anticancer properties of Persea declinata (Bl.) Kosterm bark methanolic crude extract (PDM). PDM exhibited a potent antiproliferative effect in MCF-7 human breast cancer cells, with an IC50 value of 16.68 µg/mL after 48 h of tre...

  3. Effect of Cnidium Lactone on Serum Mutant P53 and BCL-2/BAX Expression in Human Prostate Cancer Cells PC-3 Tumor-Bearing BALB/C Nude Mouse Model

    Bi, Dongbin; Yang, Mingshan; Zhao, Xia; Huang, Shiming

    2015-01-01

    Background Cnidium lactone is a natural coumarin compound that can inhibit a variety of cancer cell proliferation and induce cancer cell apoptosis. This experiment investigated the effect of cnidium lactone on molecular marker expression in prostate cancer nude mice to study its effect in inducing apoptosis. Material/Methods We randomly and equally divided 30 male BALB/C nude mice inoculated with human prostate cancer cells PC-3 into a negative control group, a cyclophosphamide group (500 mg/...

  4. Expression and significance of β-catenin,Bcl-2 and Bax in basal cell carcinoma%β-catenin和Bcl-2、Bax蛋白在皮肤基底细胞癌中的表达和意义

    黄卡特; 吕明芬; 徐云升; 邹炳辉

    2010-01-01

    目的:探讨皮肤基底细胞癌(basal cell carcinoma,BCC)中β-catenin和Bcl-2、Bax蛋白的表达与BCC发生发展的关系.方法:采用免疫组化方法检测BCC组织切片中β-catenin和Bcl -2、Bax蛋白的表达.结果:48例BCC标本中β-catenin蛋白的异常表达率为91.67%,Bcl -2、Bax蛋白的表达率分别为89.58%、62.5%,且Bcl-2与Bax蛋白的表达呈负相关.结论:β-catenin和Bcl-2、Bax蛋白的异常表达在BCC的发生、发展中起重要作用.

  5. NCBI nr-aa BLAST: CBRC-TSYR-01-0283 [SEVENS

    Full Text Available CBRC-TSYR-01-0283 ref|NP_057140.2| transmembrane BAX inhibitor motif containing 4 [...Homo sapiens] gb|EAW97159.1| transmembrane BAX inhibitor motif containing 4, isoform CRA_b [Homo sapiens] gb|AAI48529.1| Transme...mbrane BAX inhibitor motif containing 4 [synthetic construct] gb|AAI56585.1| Transmembrane BAX inhibit...or motif containing 4 [synthetic construct] dbj|BAG73822.1| transmembrane BAX inhibit

  6. DNA repair in human bronchial epithelial cells

    The purpose of this investigation was to compare the response of human cell types (bronchial epithelial cells and fibroblasts and skin fibroblasts) to various DNA damaging agents. Repair of DNA single strand breaks (SSB) induced by 5 krads of X-ray was similar for all cell types; approximately 90% of the DNA SSB were rejoined within one hour. During excision repair of DNA damage from u.v.-radiation, the frequencies of DNA SSB as estimated by the alkaline elution technique, were similar in all cell types. Repair replication as measured by BND cellulose chromatography was also similar in epithelial and fibroblastic cells after u.v.-irradiation. Similar levels of SSB were also observed in epithelial and fibroblastic cells after exposure to chemical carcinogens: 7,12-dimethylbenz[a]anthracene; benzo[a]pyrene diol epoxide (BPDE); or N-methyl-N-nitro-N-nitrosoguanidine. Significant repair replication of BPDE-induced DNA damage was detected in both bronchial epithelial and fibroblastic cells, although the level in fibroblasts was approximately 40% of that in epithelial cells. The pulmonary carcinogen asbestos did not damage DNA. DNA-protein crosslinks induced by formaldehyde were rapidly removed in bronchial cells. Further, epithelial and fibroblastic cells, which were incubated with formaldehyde and the polymerase inhibitor combination of cytosine arabinoside and hydroxyurea, accumulated DNA SSB at approximately equal frequencies. These results should provide a useful background for further investigations of the response of human bronchial cells to various DNA damaging agents

  7. Abnormal expression of c-Myc in human bronchial epithelial cells malignantly transformed by anti-BPDE

    Juan FU; Yiguo JIANG; Xuemin CHEN

    2008-01-01

    Anti-benzo[a]pyrene-7,8-diol-9,10-epoxide (anti-BPDE) is a metabolite of benzo[a]pyrene (B[a] P) and acts as a potent mutagen in mammalian systems. However, molecular mechanisms related to anti-BPDE-induced carcinogenesis are poorly understood. Here, we investigated the expression of proto-oncogene c-myc in human bronchial epithelial cells (16H BE-T) transformed by exposure to anti-BPDE. The levels ofmRNA and pro-tein of c-M yc were examined in the 16HBE-T and vehicle-treated control cells (16HBE-N) by using different meth-ods respectively, including reverse transcriptase-polymer-ase chain reaction (RT-PCR), quantitative real-time PCR (Q-PCR), western blot and immunocytochemical meth-ods. The level of c-myc mRNA appeared to be signifi-cantly increased in 16HBE-T, as compared with those of the 16H BE-N. Likewise, the expression of c-Myc protein was significantly enhanced as compared with those of the control cells. Moreover, the localization of c-Myc protein shows mainly nuclear staining in 16HBE-T. In conclu-sion, the abnormal expression of c-Myc was present in anti-BPDE malignantly transformed 16HBE cells, which may be involved in the carcinogenesis molecular mech-anism of anti-BPDE.

  8. Expression of Apoptosis Related Protein in Skin Lesions of Lichen Planus and Its Implication

    Xu'e CHEN; Yan WU; Jiawen LI; Zhixiang LIU; Qing YUE; Houjun LIU

    2008-01-01

    In order to investigate the role of Caspase-3 and Bax in the pathogenesis of lichen planus, immunohistochemistry was used to detect the expression of Caspase-3 and Bax in skin lesions of the patients with lichen planus and skin tissues of normal subjects. The results showed that positive rate of Caspase-3 and Bax expression in lichen planus were significantly higher than that in normal skins (both P<0.05). Meanwhile, there was a obvious correlation between the increase of Caspase-3 and that of Bax in lichen planus. The expression of Caspase-3 and Bax might play an important role in the development of lichen planus.

  9. NCBI nr-aa BLAST: CBRC-AGAM-01-0043 [SEVENS

    Full Text Available CBRC-AGAM-01-0043 ref|NP_071435.2| transmembrane BAX inhibitor motif containing 1 [...Homo sapiens] sp|Q969X1|TMBI1_HUMAN Transmembrane BAX inhibitor motif-containing protein 1 (Protein RECS1 ho...molog) gb|AAH26348.1| Transmembrane BAX inhibitor motif containing 1 [Homo sapiens] gb|EAW70607.1| transmembrane BAX inhibit...or motif containing 1, isoform CRA_a [Homo sapiens] gb|EAW70609.1| transme...mbrane BAX inhibitor motif containing 1, isoform CRA_a [Homo sapiens] NP_071435.2 0.13 26% ...

  10. NCBI nr-aa BLAST: CBRC-AGAM-01-0043 [SEVENS

    Full Text Available CBRC-AGAM-01-0043 ref|NP_001007714.1| transmembrane BAX inhibitor motif containing ...1 [Rattus norvegicus] gb|AAH79087.1| Transmembrane BAX inhibitor motif containing 1 [Rattus norvegicus] gb|EDL75346.1| transme...mbrane BAX inhibitor motif containing 1, isoform CRA_a [Rattus norvegicus] gb|EDL75347.1| transme...mbrane BAX inhibitor motif containing 1, isoform CRA_a [Rattus norvegicus] gb|EDL75348.1| transme...mbrane BAX inhibitor motif containing 1, isoform CRA_a [Rattus norvegicus] NP_001007714.1 0.23 28% ...

  11. Effect of strand-specific excision repair on the spectra of mutations induced by benzo[a]pyrene-diol epoxide and ultraviolet radiation in diploid human cells

    To study the effect of excision repair on the spectra of mutations induced in diploid human cells by UV and ±-7β, 8α-dihydroxy-9α,10α-epoxy- 7,8,9,10-tetrahydrobenzo[a]pyrene (BPDE), the author synchronized repair-proficient cells, treated them at the beginning of S phase or in G1 phase several hours prior to the onset of S phase, selected for thioguanine resistant cells, and determined the spectra of mutations in the coding region of the hyproxanthine(guanine)phosphoribosyl-transferase (HPRT) gene in the mutants. As a control, the spectra of mutations similarly induced in repair-deficient xeroderma pigmentosum (XP) cells were compared. There was no difference in the kinds of mutations observed in mutants derived from either cell strain treated with a particular mutagen either in S or in G1. With BPDE, the majority were G.C→T.A transversions; with UV, they were mainly G.C.→A.T transitions. The strand distribution of premutagenic lesions in mutants from repair-proficient cells treated in S or G1 differed significantly. The results strongly support the hypothesis that human cells preferentially repair UV- and BPDE-induced lesions from the transcribed strand of the HPRT gene. To test this, the rate of repair of BPDE adducts from individual strands of the HPRT gene was measured, using the UvrABC exinuclease and Southern hybridizations with strand-specific probes to detect lesions remaining. BPDE lesions were removed from the transcribed strand at a significantly faster rate than from the nontranscribed strand, consistent with my hypothesis. It was found that BPDE adducts were removed faster from either strand of the HPRT gene than from a transcriptionally inactive locus, indicating preferential repair of active genes. The results of these studies provide biochemical and biological evidence of strand-specific DNA repair of BPDE adducts in human cells

  12. DNA-protective effects of sumach (Rhus coriaria L.), a common spice: Results of human and animal studies

    Chakraborty, Asima; Ferk, Franziska; Simic, Tatjana [Institute of Cancer Research, Department of Medicine I, Medical University of Vienna, Borschkegasse 8a, A-1090 Vienna (Austria); Brantner, Adelheid [Institute of Pharmacognosy, University of Graz, Universitaetsplatz 4/I, A-8010 Graz (Austria); Dusinska, Maria [Center for Ecological Economics, Norwegian Institute for Air Research, Instituttveien 18, NO-2027 Kjeller (Norway); Kundi, Michael [Institute of Environmental Health, Center for Public Health, Medical Unviversity of Vienna (Austria); Hoelzl, Christine; Nersesyan, Armen [Institute of Cancer Research, Department of Medicine I, Medical University of Vienna, Borschkegasse 8a, A-1090 Vienna (Austria); Knasmueller, Siegfried [Institute of Cancer Research, Department of Medicine I, Medical University of Vienna, Borschkegasse 8a, A-1090 Vienna (Austria)], E-mail: siegfried.knasmueller@meduniwien.ac.at

    2009-02-10

    Sumach (Rhus coriaria L.) is widely used as a spice. The aim of this study was the investigation of its DNA-protective effects in humans and animals. Prevention of the formation of strand breaks and oxidized DNA bases as well as the protection against H{sub 2}O{sub 2}- and ({+-})-anti-benzo[a]pyrene-7,8-dihydro-diol-9,10-epoxide (BPDE)-induced DNA-damage were monitored in human lymphocytes in a placebo controlled trial (N = 8/group) with ethanolic extract of sumach (3.0 g/day, 3 days) in single cell gel electrophoresis assays. Furthermore, DNA-protective effects of sumach were monitored in different inner organs of rats under identical conditions. No alteration of DNA-migration was detectable in human lymphocytes under standard conditions, but a decrease of the tail-lengths due to formation of oxidized purines and pyrimidines (52% and 36%) was found with lesion-specific enzymes. Also damage caused by H{sub 2}O{sub 2} and BPDE was significantly reduced by 30% and 69%, respectively. The later effect may be due to induction of glutathione S-transferase (GST). After the intervention, the overall GST (CDNB) activity in plasma was increased by 40%, GST-{alpha} by 52% and GST-{pi} by 26% (ELISA). The antioxidant effects of extract are probably due to scavenging which was observed in in vitro experiments, which also indicated that gallic acid is the active principle of sumach. The animal experiments showed that sumach also causes protection in inner organs. Supplementation of the drinking water (0.02 g/kg per animal) decreased the formation of oxidized DNA bases in colon, liver, lung and lymphocytes; also after {gamma}-irradiation pronounced effects were seen.

  13. Benzo[a]pyrene diol epoxide suppresses retinoic acid receptor-β2 expression by recruiting DNA (cytosine-5--methyltransferase 3A

    Xu Xiao-Chun

    2010-04-01

    Full Text Available Abstract Tobacco smoke is an important risk factor for various human cancers, including esophageal cancer. How benzo [a]pyrene diol epoxide (BPDE, a carcinogen present in tobacco smoke as well as in environmental pollution, induces esophageal carcinogenesis has yet to be defined. In this study, we investigated the molecular mechanism responsible for BPDE-suppressed expression of retinoic acid receptor-beta2 (RAR-β2 in esophageal cancer cells. We treated esophageal cancer cells with BPDE before performing methylation-specific polymerase chain reaction (MSP to find that BPDE induced methylation of the RAR-β2 gene promoter. We then performed chromatin immunoprecipitation (ChIP assays to find that BPDE recruited genes of the methylation machinery into the RAR-β2 gene promoter. We found that BPDE recruited DNA (cytosine-5--methyltransferase 3 alpha (DNMT3A, but not beta (DNMT3B, in a time-dependent manner to methylate the RAR-β2 gene promoter, which we confirmed by reverse transcription-polymerase chain reaction (RT-PCR analysis of the reduced RAR-β2 expression in these BPDE-treated esophageal cancer cell lines. However, BPDE did not significantly change DNMT3A expression, but it slightly reduced DNMT3B expression. DNA methylase inhibitor 5-aza-2'-deoxycytidine (5-Aza and DNMT3A small hairpin RNA (shRNA vector antagonized the effects of BPDE on RAR-β2 expressions. Transient transfection of the DNMT3A shRNA vector also antagonized BPDE's effects on expression of RAR-β2, c-Jun, phosphorylated extracellular signal-regulated protein kinases 1/2 (ERK1/2, and cyclooxygenase-2 (COX-2, suggesting a possible therapeutic effect. The results of this study form the link between the esophageal cancer risk factor BPDE and the reduced RAR-β2 expression.

  14. DNA-protective effects of sumach (Rhus coriaria L.), a common spice: Results of human and animal studies

    Sumach (Rhus coriaria L.) is widely used as a spice. The aim of this study was the investigation of its DNA-protective effects in humans and animals. Prevention of the formation of strand breaks and oxidized DNA bases as well as the protection against H2O2- and (±)-anti-benzo[a]pyrene-7,8-dihydro-diol-9,10-epoxide (BPDE)-induced DNA-damage were monitored in human lymphocytes in a placebo controlled trial (N = 8/group) with ethanolic extract of sumach (3.0 g/day, 3 days) in single cell gel electrophoresis assays. Furthermore, DNA-protective effects of sumach were monitored in different inner organs of rats under identical conditions. No alteration of DNA-migration was detectable in human lymphocytes under standard conditions, but a decrease of the tail-lengths due to formation of oxidized purines and pyrimidines (52% and 36%) was found with lesion-specific enzymes. Also damage caused by H2O2 and BPDE was significantly reduced by 30% and 69%, respectively. The later effect may be due to induction of glutathione S-transferase (GST). After the intervention, the overall GST (CDNB) activity in plasma was increased by 40%, GST-α by 52% and GST-π by 26% (ELISA). The antioxidant effects of extract are probably due to scavenging which was observed in in vitro experiments, which also indicated that gallic acid is the active principle of sumach. The animal experiments showed that sumach also causes protection in inner organs. Supplementation of the drinking water (0.02 g/kg per animal) decreased the formation of oxidized DNA bases in colon, liver, lung and lymphocytes; also after γ-irradiation pronounced effects were seen

  15. Láminas delgadas de (Bi0.5Na0.5)1-xBaxTiO3 en torno a la frontera de fase morfotrópica preparadas por métodos de depósito químico y fotoquímico de disoluciones

    Pérez-Mezcua, Dulce

    2016-01-01

    Tesis presentada para aspirar al grado de Doctor por la Universidad de Navarra. El presente trabajo ha sido realizado en los Departamentos de Química de la Facultad de Ciencias de la Universidad de Navarra y de Materiales para las Tecnologías de la Información del Instituto de Ciencia de Materiales de Madrid (ICMM-CSIC).

  16. Study on expression of p53、 bcl-2、 bax and PCNA in retinoblastoma%视网膜母细胞瘤p53、bcl-2、bax及增殖细胞核抗原的表达意义

    李琳; 雷霍; 杨慧

    2002-01-01

    目的了解视网膜母细胞瘤(retinoblastoma,Rb)p53、bcl-2、bax和增殖细胞核抗原表达与Rb及其分化程度的关系.方法用免疫组化检测21例Rb和6例正常视网膜常规石蜡标本进行p53、bcl-2、bax和PCNA表达.结果bc1-2、bax和PCNA在Rb中的表达较高,与正常视网膜相比有显著性差异,分化性Rb组织bcl-2阳性表达率较高,与未分化型Rb组织相比有显著性差异.p53、bcl-2、bax在Rb中的表达与PCNA的表达有显著相关性.结论 Rb的发生与多个细胞凋亡调控基因异常有关,测定和分析Rb的bcl-2、PCNA、bax和表达,对于Rb的诊断和分化程度有重要意义.

  17. 小熊猫小脑皮层的组织结构及RT-97、KGF和Bax蛋白在小脑皮层的表达%Histological Structure and Expression of RT-97, KGF and Bax Proteins in the Cerebellar Cortex of Ailurus fulgens

    高先军; 王昱; 俞诗源; 王悦; 郭婷婷

    2009-01-01

    为了解小熊猫(Ailurus fulgens)小脑皮层的结构特征,观察神经丝蛋白抗体RT-97、角质细胞生长因子(KGF)及Bax蛋白在小脑皮层中的表达,利用组织学方法和免疫组织化学方法观察了小熊猫小脑皮层的显微结构,检测了RT-97、KGF和Bax蛋白的表达.结果表明,小脑皮层从外向内依次可分为分子层、Purkinje细胞层、颗粒层3层.RT-97在小熊猫小脑皮层Purkinje细胞层、颗粒层中神经细胞的轴突、分子层中颗粒细胞的轴突及小脑髓质中有阳性表达;KGF在小脑皮层分子层、Purkinje细胞层和颗粒细胞层及髓质中均有阳性表达;Bax蛋白在小脑皮层分子层、Purkinje细胞层和颗粒细胞层中有阳性表达.RT-97、KGF和Bax蛋白在小脑皮层神经结构的构筑中可能发挥着不同的功能.

  18. Ki67和Bax在局部晚期宫颈癌新辅助化疗前后表达的意义%The significance of Ki67 and Bax expressions before and after neoadjuvant chemotherapy in locally advanced cervical carcinoma

    游艳琴; 宋磊

    2006-01-01

    目的:探讨Ki67、Bax与宫颈癌新辅助化疗疗效和预后之间的关系.方法:在新辅助化疗(neoadjuvant chemotherapy,NACT)前后用免疫组化方法检测23例局部晚期宫颈癌组织Ki67和Bax的表达.结果:临床总有效率为82.61%.化疗有反应者NACT前后Ki67、Bax表达有统计学差异(P<0.05);无反应者NACT前后Ki67、Bax表达无统计学差异(P>0.05).不同病理类型和临床分期患者NACT前后Ki67和Bax变化值无显著性差异;不同病理分级患者NACT前后Bax变化值有统计学差异(P<0.05).NACT前Ki67 LI>33%和Ki67 LI<33%患者的平均无瘤生存期分别为48.6个月和19个月(P<0.05).结论:Ki67、Bax与化疗疗效显著相关.Ki67、Bax有可能作为判断化疗疗效和预后的指标.

  19. 苯扎贝特对ox-LDL诱导内皮细胞凋亡基因Bcl-2/Bax的影响%Effects of bezafibrate on apoptosis gene Bcl-2/Bax in cultured endothelial cells induced by ox-LDL

    申晓彧; 薛丽霞; 屈巧芳; 曾秋棠

    2008-01-01

    目的 观察苯扎贝特对氧化型低密度脂蛋白(ox-LDL)诱导人脐静脉内皮细胞凋亡基因Bcl-2/Bax的影响. 方法 体外培养人脐静脉内皮细胞(HUVECs),根据实验要求分为正常对照组、ox-LDL组、低浓度苯扎贝特(50μmol/L)组、中浓度苯扎贝特(100 μmol/L)组、高浓度苯扎贝特(200μmol/L)组.RT-PCR观察各组凋亡基因Bcl-2、Bax及Bd-2/BaxmRNA的变化. 结果 与正常对照组比较,ox-LDL组凋亡基因Bcl-2表达下降(P<0.05),凋亡基因Bax表达增加(P<0.05),Bcl-2/Bax比值下降(P<0.05);不同浓度苯扎贝特组与ox-LDL组比较,Bcl-2表达增加(P<0.05),Bax表达降低(P<0.05),Bcl-2/Bax上调(P<0.05),且呈浓度效应依赖关系. 结论 ox-LDL可引起内皮细胞抗凋亡基因Bcl-2表达下调,凋亡基因Bax表达上调,Bcl-2和Bax比值下降,从而引起内皮细胞凋亡增加;苯扎贝特可通过上调Bcl-2与Bax的比值抑制ox-LDL引起的内皮细胞凋亡,起到抗动脉粥样硬化作用.

  20. Obtaining a mutant of Bacillus amyloliquefaciens xylanase A with improved catalytic activity by directed evolution.

    Xu, Xin; Liu, Ming-Qi; Huo, Wen-Kang; Dai, Xian-Jun

    2016-05-01

    This study aimed to obtain xylanase exhibiting improved enzyme properties to satisfy the requirements for industrial applications. The baxA gene encoding Bacillus amyloliquefaciens xylanase A was mutated by error-prone touchdown PCR. The mutant, pCbaxA50, was screened from the mutant library by using the 96-well plate high-throughput screening method. Sequence alignment revealed the identical mutation point S138T in xylanase (reBaxA50) produced by the pCbaxA50. The specific activity of the purified reBaxA50 was 9.38U/mg, which was 3.5 times higher than that of its parent expressed in Escherichia coli BL21 (DE3), named reBaxA. The optimum temperature of reBaxA and reBaxA50 were 55°C and 50°C, respectively. The optimum pH of reBaxA and reBaxA50 were pH 6 and pH 5, respectively. Moreover, reBaxA50 was more stable than reBaxA under thermal and extreme pH treatment. The half-life at 60°C and apparent melting temperature of reBaxA50 were 9.74min and 89.15°C, respectively. High-performance liquid chromatography showed that reBaxA50 released xylooligosaccharides from oat spelt, birchwood, and beechwood xylans, with xylotriose as the major product; beechwood xylan was also the most thoroughly hydrolyzed. This study demonstrated that the S138T mutation possibly improved the catalytic activity and thermostability of reBaxA50. PMID:26992794

  1. Quantitative expression analysis and prognostic significance of the BCL2-associated X gene in nasopharyngeal carcinoma: a retrospective cohort study

    Nasopharyngeal carcinoma (NPC) is a highly metastatic epithelial malignancy showing high prevalence in Southeast Asia and North Africa. The BCL2-associated X (BAX) gene encodes the most important pro-apoptotic member of the BCL2 family. We have recently shown that BCL2 and BCL2L12, two other members of the same apoptosis-related family, possess significant prognostic value in NPC. The objective of the current study was to analyze BAX mRNA expression in nasopharyngeal biopsies of NPC patients, and to assess its prognostic potential in this disease. Total RNA was isolated from 88 malignant and 9 hyperplastic nasopharyngeal biopsies, resected from Tunisian patients. After cDNA synthesis by reverse transcription of polyadenylated RNA, BAX mRNA expression was analyzed using a highly sensitive quantitative real-time polymerase chain reaction (qRT-PCR) method. Lower BAX mRNA levels were detected in NPC biopsies than in hyperplastic nasopharyngeal samples. BAX mRNA expression status was associated with low tumor extent, negative regional lymph node status, and absence of distant metastases. Kaplan-Meier survival analysis demonstrated that patients with BAX mRNA-positive NPC have significantly longer disease-free survival (DFS) and overall survival (OS). In accordance with these findings, Cox regression analysis revealed that BAX mRNA expression can be considered as a favorable prognostic indicator of DFS and OS in NPC, independent of their gender, age, tumor histology, tumor extent, and nodal status. Furthermore, NPC patients without distant metastases are less likely to relapse when their primary tumor is BAX mRNA-positive, compared to metastasis-free patients with a BAX-negative nasopharyngeal malignancy. This is the first study examining the potential clinical utility of BAX as a prognostic tumor biomarker in NPC. We provide evidence that BAX mRNA expression can be considered as an independent favorable prognostic indicator of DFS and OS in NPC

  2. Platelet apoptosis by cld-induced glycoportein Ibα clustering

    van der Wal, Dianne E; Du, V X; Lo, KS; Rasmussen, Jan Trige; Verhoef, S; Akkerman, J W N

    2010-01-01

    -dephosphorylation and activation. This initiates pro-apoptosis changes in Bax/Bcl-xL and Bax-translocation to the mitochondria, inducing cytochrome c release. The result is activation of caspase-9, which triggers phosphatidylserine exposure and platelet phagocytosis by macrophages. Responses are prevented by N-acetyl-d-glucosamine...

  3. Apoptosis of dermal fibroblasts in wounds combined with radiation injury and its mechanisms

    Objective: To study apoptosis of dermal fibroblasts in wounds combined with total body irradiation (TBI) injury and its mechanisms. Methods: The authors determined the amount of dermal fibroblasts in wounds of rats using histological method, observed apoptosis of dermal fibroblasts in wounds by transmission electron microscope and TUNEL assay, and detected the expression of Bcl-2 and Bax in wounds with immunohistochemistry method. Results: The ratio of fibroblastic apoptosis in wounds combined with TBI injury significantly increased as compared with that of simple incision injury. The contents of Bax in wounds combined with TBI injury were significantly higher than those of simple incision injury, and the contents of Bcl-2 and Bcl-2/Bax ratio in the former were significantly lower. Conclusion: Apoptosis of dermal fibroblasts in wounds combined with TBI injury increases significantly, which causes reduction of the amount of fibroblasts. Abnormalities of Bcl-2, Bax and Bcl-2/Bax ratio are important reasons of the increase of fibroblastic apoptosis

  4. Design and evaluation of new chemotherapeutics of aloe-emodin (AE) against the deadly cancer disease: an in silico study.

    Mulakayala, Chaitanya; Banaganapalli, Babajan; Mulakayala, Naveen; Pulaganti, Madhusudana; C M, Anuradha; Chitta, Suresh Kumar

    2013-05-23

    The Bcl-2 family proteins include pro- and antiapoptotic factors acting as critical arbiters of apoptotic cell death decisions in most circumstances. Evasion of apoptosis is one of the hallmarks of cancer, relevant to tumorigenesis as well as resistance to cytotoxic drugs, and deregulation of Bcl-2 proteins was observed in many cancers. Since Bax-mediated induction of apoptosis is a crucial mechanism in cancerous cells, we aimed at conducting in silico analysis on Bax in order to predict the possible interactions for anticancer agents. The present report depicts the binding mode of aloe-emodin and its structurally modified derivatives onto Bax. The structural information about the binding site of Bax for docked compounds obtained from this study could aid in screening and designing new anticancer agents or selective inhibitors for chemotherapy against Bax. PMID:24432130

  5. Genotoxicity of soluble and particulate cadmium compounds: impact on oxidative DNA damage and nucleotide excision repair.

    Schwerdtle, Tanja; Ebert, Franziska; Thuy, Christina; Richter, Constanze; Mullenders, Leon H F; Hartwig, Andrea

    2010-02-15

    Water-soluble and particulate cadmium compounds are carcinogenic to humans. While direct interactions with DNA are unlikely to account for carcinogenicity, induction of oxidative DNA damage and interference with DNA repair processes might be more relevant underlying modes of action (recently summarized, for example, in Joseph , P. (2009) Tox. Appl. Pharmacol. 238 , 271 - 279). The present study aimed to compare genotoxic effects of particulate CdO and soluble CdCl(2) in cultured human cells (A549, VH10hTert). Both cadmium compounds increased the baseline level of oxidative DNA damage. Even more pronounced, both cadmium compounds inhibited the nucleotide excision repair (NER) of BPDE-induced bulky DNA adducts and UVC-induced photolesions in a dose-dependent manner at noncytotoxic concentrations. Thereby, the uptake of cadmium in the nuclei strongly correlated with the repair inhibition of bulky DNA adducts, indicating that independent of the cadmium compound applied Cd(2+) is the common species responsible for the observed repair inhibition. Regarding the underlying molecular mechanisms in human cells, CdCl(2) (as shown before by Meplan, C., Mann, K. and Hainaut, P. (1999) J. Biol. Chem. 274 , 31663 - 31670 ) and CdO altered the conformation of the zinc binding domain of the tumor suppressor protein p53. In further studies applying only CdCl(2), cadmium decreased the total nuclear protein level of XPC, which is believed to be the principle initiator of global genome NER. This led to diminished association of XPC to sites of local UVC damage, resulting in decreased recruitment of further NER proteins. Additionally, CdCl(2) strongly disturbed the disassembly of XPC and XPA. In summary, our data indicate a general nucleotide excision repair inhibition by cadmium compounds, which is most likely caused by a diminished assembly and disassembly of the NER machinery. These data reveal new insights into the mechanisms involved in cadmium carcinogenesis and provide further

  6. Apoptosis induced by radionuclide 153Sm and expression of relevant genes in three different cancer cells

    To study apoptosis of PC-3, ER-75-30 and A549 cells induced by radionuclide 153Sm and the expression of bcl-2, bax in apoptosis cells, MTT assay was used to detect the anti-tumor effect, light microscope, transmission electron microscope, flow cytometer were used to detect apoptosis, while image analysis was used to detect the expression of bcl-2 and bax. 153Sm showed anti-tumor effect and could induce tumor cell apoptosis. Both bcl-2 and bax played an important role in apoptosis. Different kind of cells had different sensitivity to 153Sm

  7. Lattice instabilities and superconductivity in La2-x(Ba,Sr)xCuO4

    In this paper, the authors present the result of two experiments on the anomalous physical properties of La2-xBaxCuO4 with x = 0.125. The suppression of a low-temperature tetragonal phase by introduction of Sr in the La2-xBaxCuO4 correlates well with the recovery of 30-K superconductivity. Hydrogenation of La2-xBaxCuO4 revealed that the carrier density, rather than the amount of divalent ions, is an essential parameter which governs the anomaly

  8. Effect of propofol pretreatment on apoptosis in rat brain cortex after focal cerebral ischemia and reperfusion

    Haiyan Xu; Chengwei Zhang; Chunxiao Zhang

    2011-01-01

    The present study aimed to observe cortical expression of Bcl-2 and Bax, cysteine-dependent aspartate directed proteases-3 activity and apoptotic cell death in a rat model of middle cerebral artery occlusion pretreated with propofol. Results showed that, propofol pretreatment significantly reduced oxidative stress levels and attenuated neuronal apoptosis in the cortex of rats. Propofol pretreatment upregulated Bcl-2 expression, and downregulated Bax expression and cysteine-dependent aspartate directed proteases-3 activity. These findings indicate that propofol pretreatment inhibits cell apoptosis during focal cerebral ischemia/reperfusion injury. This neuroprotective effect is most likely achieved through the Bcl-2/Bax/cysteine-dependent aspartate directed proteases-3 pathway.

  9. SwissProt search result: AK070925 [KOME

    Full Text Available AK070925 J023070O07 (Q8BJZ3) Transmembrane BAX inhibitor motif containing protein 1 (RECS1 prote ... in) (Responsive to centrifugal force ... and shear stress gene 1 protein) TMBI1_MOUSE 6e-11 ...

  10. SwissProt search result: AK073781 [KOME

    Full Text Available AK073781 J033068H10 (Q8BJZ3) Transmembrane BAX inhibitor motif containing protein 1 (RECS1 prote ... in) (Responsive to centrifugal force ... and shear stress gene 1 protein) TMBI1_MOUSE 1e-15 ...

  11. SwissProt search result: AK102207 [KOME

    Full Text Available AK102207 J033087H03 (Q8BJZ3) Transmembrane BAX inhibitor motif containing protein 1 (RECS1 prote ... in) (Responsive to centrifugal force ... and shear stress gene 1 protein) TMBI1_MOUSE 1e-15 ...

  12. SwissProt search result: AK070043 [KOME

    Full Text Available AK070043 J023039E14 (Q8BJZ3) Transmembrane BAX inhibitor motif containing protein 1 (RECS1 prote ... in) (Responsive to centrifugal force ... and shear stress gene 1 protein) TMBI1_MOUSE 3e-11 ...

  13. SwissProt search result: AK242300 [KOME

    Full Text Available AK242300 J075192P11 (Q8BJZ3) Transmembrane BAX inhibitor motif containing protein 1 (RECS1 prote ... in) (Responsive to centrifugal force ... and shear stress gene 1 protein) TMBI1_MOUSE 9e-15 ...

  14. SwissProt search result: AK069449 [KOME

    Full Text Available AK069449 J023020E21 (Q8BJZ3) Transmembrane BAX inhibitor motif containing protein 1 (RECS1 prote ... in) (Responsive to centrifugal force ... and shear stress gene 1 protein) TMBI1_MOUSE 2e-15 ...

  15. EXPRESSION AND SIGNIFICANCE OF bcl-2 FAMILY IN AMELOBLASTONA

    WANG Jie; MA Jie; ZHONG Ming; LIU Jing-dong

    2006-01-01

    Objective: To study the expression of bcl-2 and bax in human ameloblastoma (AB), and investigate the role of apoptosis in genesis and development of AB and the relation of apoptosis with the clinic biological characteristics of AB. Methods:BCL-2 and BAX proteins were detected in 75 cases of AB (primary AB 31 cases, recurrent AB 37 cases, malignant AB 7cases) by S-P method. Oral normal mucosa (NOM) and Odontogenic kerotosyst (OKC) were used as controls. Bcl-2 and bax mRNA in 20 cases of AB, 12 cases of OKC were detected by in situ hybridization. Results: The positive ratio of BCL-2protein was 88.0% ( 66/75 ) in AB, 74.3% (26/35) in OKC and 44.4% (4/9) in NOM, respectively (P<0.001). BCL-2 protein was expressed in peripheral cells and a few scattered stellate-shape cells in AB. The positive ratio of BAX protein was 74.7%(56/75)in AB, 65.7%(23/35)in OKC and 77.8%(7/9) in NOM, respectively (P<0.001). BAX protein was expressed in peripheral cells and stellate-shape cells with similar intensity. BCL-2 expression increased in recurrent and AB canceration(P<0.01), while for BAX expression, the positive ratio was higher in recurrent AB, but lower than that of malignant AB. A moderate negative correlation between BCL-2 and BAX protein was found (rk=-0.331, P<0.001).Conclusion: AB has much more apoptosis-inhibiting protein than apoptosis- accelarating protein. Apoptosis plays an important role in genesis, development of AB. The fashion and intensity of bcl-2 and bax expression were different in various tissues and in benign or malignant AB.

  16. Apoptosis regulates ipRGC spacing necessary for rods and cones to drive circadian photoentrainment

    Chen, Shih-Kuo; Chew, Kylie S.; McNeill, David S.; Keeley, Patrick W.; Ecker, Jennifer L.; Mao, Buqing Q.; Pahlberg, Johan; Kim, Bright; Lee, Sammy C. S.; Fox, Michael; Guido, William; Wong, Kwoon Y.; Sampath, Alapakkam P.; Reese, Benjamin E.; Kuruvilla, Rejji

    2013-01-01

    The retina consists of ordered arrays of individual types of neurons for processing vision. Here we show that such order is necessary for intrinsically photosensitive retinal ganglion cells (ipRGCs) to function as irradiance detectors. We found that during development, ipRGCs undergo proximity-dependent Bax-mediated apoptosis. Bax mutant mice exhibit disrupted ipRGC spacing and dendritic stratification with an increase in abnormally localized synapses. ipRGCs are the sole conduit for light in...

  17. Production of translationally cold barium monohalide ions

    DePalatis, M. V.; Chapman, M.S.

    2013-01-01

    We have produced sympathetically cooled barium monohalide ions BaX$^+$ (X = F, Cl, Br) by reacting trapped, laser cooled Ba$^+$ ions with room temperature gas phase neutral halogen-containing molecules. Reaction rates for two of these (SF$_6$ and CH$_3$Cl) have been measured and are in agreement with classical models. BaX$^+$ ions are promising candidates for cooling to the rovibrational ground state, and our method presents a straightforward way to produce these polar molecular ions.

  18. Different Expressions of HIF-1α, Bcl-2 and Baxin DU145 Prostate Cancer Cells Transplanted in Nude Mouse between X-Ray and Neutron Irradiation

    To investigate the radiobiologic effects of neutron and X-ray irradiation on DU-145 prostate carcinoma cells by identifying the differences of HIF-1α expression and apoptosis. Nude mice were injected with the human prostate cancer cell line, DU-145, and then irradiated with 2 Gy and 10 Gy X-rays, or 0.6 Gy and 3.3 Gy neutrons, respectively. The mice were sacrificed at 24 hours and 120 hours after irradiation. The expression levels of HIF-1α, Bcl-2 and Bax were compared with immunohistochemical staining and western blotting. The apoptotic indexes were compared with the Terminal deoxynucleotidyl biotin-dUTP nick and labeling (TUNEL) assay. At day 1, HIF-1α and Bcl-2 expression decreased, while Bax expression and the number of TUNEL positive cells increased in neutron irradiated groups for the control and X-ray irradiated groups. The Bcl-2/Bax ratio was significantly lower in the neutron irradiated groups regardless of dose (p=0.001). The same pattern of the differences in the expressions of the HIF-1α, Bcl-2, Bax, Bcl-2/Bax ratio, and apoptotic indexes were indentified at day 5. HIF-1α expression was related with Bcl-2 (p=0.031), Bax (p=0.037) expressions and the apoptotic indexes (p=0.016) at day 5. Neutron irradiation showed a decrease in HIF-1α, Bcl-2 expression, and Bcl-2/Bax ratio, but increased Bax expression regardless of dose. This study suggests that the differences radiobiological responses between photon and neutron irradiation may be related to different HIF-1α expression and subsequent apoptotic protein expressions

  19. Efficacy of a commercial polymerase chain reaction-based assay for detection of Salmonella spp. in animal feeds.

    Maciorowski, K G; Pillai, S D; Ricke, S C

    2000-10-01

    Salmonellosis is a cyclic problem in the food industry, to which animal feed has been contributory. Current conventional methods of Salmonella spp. detection require 96 h for detection and confirmation. With modern and just-in-time production schedules, a 96-h hold represents a significant expense in storage and decontamination. The commercially available assay, 'BAX for Screening/Salmonella' (BAX), is based on the principle of the polymerase chain reaction and may represent a significant decrease in assay time. Seven fresh feed formulations, two fresh feed ingredients, seven stored feeds and two stored feed ingredients were artificially contaminated with a primary poultry isolate of Salmonella typhimurium and analysed by conventional and BAX methodology. The results of BAX agreed with conventional plating results for 16 of 18 samples spiked with 1200 cfu 10 g(-1) of feed and 13 of 18 samples spiked with 40 cfu 10 g(-1) of feed. Indigenous Salmonella spp. were detected in five of eight samples of poultry diets by conventional methods. With BAX, Salmonella spp. could not be detected in any of the samples after only 7 h of enrichment but could be detected in two dietary samples after 13 h of enrichment and four dietary samples after 24 h of enrichment. Specific sequences of salmonella DNA that were extracted from poultry diets could be detected with BAX. PMID:11054177

  20. Prognostic Significance of Apoptosis Related Gene Family bcl-2 in Human Breast Cancer

    2000-01-01

    To study the prognostic effect of bcl-2 oncogene and its gene family members bax, bcl-x expression in breast cancer patients. Methods: expression of bcl-2, bax proteins in 91 human breast cancer tissue sections were studied by immunohistochemical method. Bcl-x1 mRNA expression in frozen tissues from 16 breast cancer patients were detected using Northern blot method. Results: bcl-2 protein positivity was found in 60/91 (65.9%) patients, and bax positivity 59/91 (64.8%). Bcl-2 and bax expression levels were associated with apoptotic index(AI), histological grade, axillary lymph node metastasis, postoperative local recurrence and metastasis. Bcl-2 expression was related to ER positivity. In univariate analysis for disease free survival (DFS), bcl-2 and bax protein levels, and Al were all found to have prognostic value. The result of Cox's model multivariate analysis showed that bcl-2 protein level was an independent prognostic factor. In 16 frozen breast cancer tissues, 8/16(50%) had higher level of bcl-x1 mRNA, which showed correlation with bcl-2 protein expression and axillary lymph node metastasis. Conclusion: The findings indicate that dysregulated expressions of bcl-2, bax and bcl-x1 apoptosis-related genes, suggestive of serious deregulation of apoptotic process, may contribute to the biologic aggressiveness of breast cancer. Bcl-2 protein is an independent indicator of prognosis in breast cancer patients.

  1. Relationship of angiogenic and apoptotic activities in soft-tissue sarcoma

    Thin Thin Win

    2014-01-01

    Full Text Available Introduction: Angiogenesis and apoptosis play an essential role in tumor development and progression. Previous studies on apoptosis and angiogenesis of soft-tissue sarcoma (STS were done separately. This is the first study of the relationship between apoptotic and angiogenic activity. Correlation of expression of anti-apoptotic protein (Bcl-2 and pro-apoptotic protein (Bax in the tumor cells (TCs with their expression in endothelial cell (EC of the tumor blood vessels in STS were also carried out. Materials and Methods: 101 cases of STS; consisting liposarcoma, malignant fibrous histiocytoma, synovial sarcoma, fibrosarcoma, leiomyosarcoma, rhabdomyosarcoma and malignant peripheral nerve sheath tumor; were collected and immunohistochemical reaction of vascular endothelial growth factor (VEGF, Bcl-2 and Bax were examined. Results: Higher Bax expression in TCs (54.5% was seen compared to Bcl-2 expression (44.6%. There was a significant association between Bcl-2 and Bax in TCs with ECs. Significant association was also seen between histological types of STS with Bcl-2 expression; however not with Bax expression. There was an association between VEGF and Bax with high VEGF expression and weak Bax expression. However, VEGF expression was not associated with Bcl-2 expression and histological types. Conclusion: This study supports the role of ECs of tumor blood vessels and apoptosis of TCs in tumor management. Increased angiogenesis may inhibit apoptosis of TCs and lead to tumor growth. Therefore, inhibition of ECs survival or activation of ECs death is promising prospect for tumor therapy. Immunohistochemical antibodies in this study might be potential useful marker for the prognosis of STS.

  2. Hypergravity modifies the signal transduction of ionizing radiation through p53

    Okaichi, Kumio; Usui, Aya; Okumura, Yutaka [Nagasaki Univ. (Japan). Atomic Disease Inst.; Ohnishi, Takeo [Nara Medical Univ., Kashihara (Japan)

    2002-12-01

    To determine the possible effect of hypergravity to modify the signal transduction of ionizing radiation, we analyzed the accumulation of p53 and the expression of p53-dependent genes, Waf-1 and Bax, using the western blot analysis. Hypergravity (20 x g) induced the accumulation of p53 in the human glioblastoma cell line A172 after 3 h of incubation. Low-dose (0.5 Gy) irradiation to the cells accumulated p53 1.5 h after irradiation, and induced Waf-1 and Bax. Under the condition of hypergravity (20 x g), the peak of p53 accumulation was shifted from 1.5 h to 3 h after irradiation, and the inductions of Waf-1 and Bax were suppressed entirely. These results indicate that hypergravity modifies the signal transduction of ionizing radiation through p53 in the cells. (author)

  3. Studies on the mechanism of apoptasis by radon in lung tissue of mice

    Objective: To study the mechanism of apoptasis by radon in lung tissue of mice. Methods: Male BALB/c mice were exposed to radon with the cumulative dose of 0.02, 30 or 60 working level month (WLM) respectively, and then were raised for different time (24 h, 30 d or 90 d). Apoptosis was detected by terminal deoxynucleotidy transferase-mediated dUTP-biotin nick end labeling (TUNEL). The expression of p 53 and Bcl-2/Bax protein was observed by immunohistochemistry. Results: As compared with the control group, the apoptotic index in lung tissue increased along with the increasing of expose dose and the raise time. The protein expression of p 53 increased significantlyin the 30 d and 90 d groups. But Bcl-2/Bax expression decreased. Conclusions: The apoptosis by radon in lung tissue of mice had close relationship with p 53 and.Bcl-2/Bax pathway. (authors)

  4. Molecular Mechanism of Bovine Trabecular Meshwork Cells Apoptosis Induced by Dexamethasone and Protection by Pilocarpine

    Yajuan Gu; Shujun Zeng; Pengxin Qiu; Yuping Wu; Dawei Peng; Guangmei Yan

    2005-01-01

    Purpose: To study the molecular mechanism of trabecular meshwork cells apoptosis induced by dexamethasone and the protection of pilocarpine.Methods: Determining mRNA expression with reverse transcription-polymerase chain reaction (RT-PCR), protein expression with Western blots and the percentage of apoptotic cells with fluorescent microscopy.Results: Dexamethasone up-regulated Fas proteins and affected Bax, caspase-8 and caspase-9 proteins in an action of first decrease then increase. Pre-treatment with pilocarpine decreased the four proteins expression, which were increased by dexamethasone. Pilocarpine self could decrease pro-apoptotic factors Bax, caspase-8 and caspase-9 proteins expression.Conclusion: Fas/FasL pathway participated in apoptotic process induced by dexamethasone in trabecular meshwork cells and the process was probably related with both caspase-8 and caspase-9 pathways. Pilocarpine protected the cells against apoptosis through down-regulating Fas, Bax, caspase-8 and caspase-9 proteins expression.

  5. Effect of Yiqi Bushen prescription on hippocampal neuronal apoptosis in diabetic rats

    Deshan Liu; Weiwei Lin; Wei Gao; Ping Chang; Wei Li

    2011-01-01

    This study investigated the neuroprotective effect of Yiqi Bushen prescription (YQBS, supplementing qi and tonifying kidney) on neuronal cell apoptosis. Following YQBS treatment, the number of surviving hippocampal neurons increased, anti-apoptotic Bcl-2 expression increased and pro-apoptotic Bax expression decreased. In addition, diabetic rats exhibited improved learning and memory. YQBS treatment also increased Bcl-2 mRNA expression and the ratio of Bcl-2/Bax, but decreased levels of hypoxia-inducible factor-1α mRNA and Bax mRNA expression after high-glucose/hypoxia-induced injury. Results demonstrated that YQBS inhibited hippocampal neuronal apoptosis by decreasing hypoxia-inducible factor-1α expression and increasing Bcl-2 expression, thereby improving cognitive impairment in diabetic rats.

  6. Kanglaite combined Gemcitabine inhibits growth of nude mouse subcutaneous transplantation tumor of human PC-3 pancreatic cancer cell

    WANG Wei; JIN Jian-guang; QIN Zhao-yin

    2005-01-01

    Objective:To study the mechanisms of pancreatic cancer treatment with Kanglaite combined Gemcitabine by investigating the relationship between the apoptosis and the expression of bcl-2, Bax and VEGF in pancreatic cancer cells.Methods:Nude mouse subcutaneous transplantation tumor model of Human PC-3 pancreatic cancer was established; the expressions of bcl-2, Bax and VEGF of transplantation tumor cell were determined; the earlier apoptosis rate of pancreatic cancer cell and the gross tumor volume were determined. Results:Kanglaite combined Gemcitabine remarkably decreased the protein expression of bcl-2,raised the expression of Bax,increased the apoptosis rate of the pancreatic cancer and contract the gross tumor volume. Kanglaite greatly decreased the protein expression of VEGF of the tumor cell. Conclusion:Therapeutic efficacy of Kanglaite combined Gemcitabine is far better than separate use of the two medicines in the pancreatic cancer transplantation tumor treatment.

  7. Protective Roles of Sodium Selenite against Aflatoxin B1-Induced Apoptosis of Jejunum in Broilers

    Xi Peng

    2014-12-01

    Full Text Available The effects of aflatoxin B1 (AFB1 exposure and sodium selenite supplementation on cell apoptosis of jejunum in broilers were studied. A total of 240 one-day-old male AA broilers were randomly assigned four dietary treatments containing 0 mg/kg of AFB1 (control, 0.3 mg/kg AFB1 (AFB1, 0.4 mg/kg supplement Se (+ Se and 0.3 mg/kg AFB1 + 0.4 mg/kg supplement Se (AFB1 + Se, respectively. Compared with the control broilers, the number of apoptotic cells, the expression of Bax and Caspase-3 mRNA were significantly increased, while the expression of Bcl-2 mRNA and the Bcl-2/Bax ratio were significantly decreased in AFB1 broilers. The number of apoptotic cells and the expression of Caspase-3 mRNA in AFB1 + Se broilers were significantly higher than those in the control broilers, but significantly lower than those in AFB1 broilers. There were no significant changes in the expression of Bax mRNA between AFB1 + Se and control broilers; the expression of Bcl-2 mRNA and the Bcl-2/Bax ratio in AFB1 + Se broilers were significantly lower than those in the control broilers, but significantly higher than those in AFB1 broilers. In conclusion, 0.3 mg/kg AFB1 in the diet can increase cell apoptosis, decrease Bcl-2 mRNA expression, and increase of Bax and Caspase-3 mRNA expression in broiler’s jejunum. However, supplementation of dietary sodium selenite at the concentration of 0.4 mg/kg Se may ameliorate AFB1-induced apoptosis by increasing Bcl-2 mRNA expression, and decreasing Bax and Caspase-3 mRNA expression.

  8. On the structural properties and superconductivity of room-temperature chemically oxidized La2-xBaxCuO4+y (0<=x<=0.15)

    Rial, C.; Moran, E.; Alario-Franco, M.A.; Amador, U.; Andersen, N.H.

    The insertion of oxygen within the structure of La2-xBaxCuO4+y (x less than or equal to 0.15), by means of room-temperature chemical oxidation, modifies both the physical and the structural features of these materials, Concerning the superconducting properties, the extra oxygen gives rise to an...... differences concerning the modifications induced by this oxidation process in the present series of La2-xBaxCuO4+y compounds and in equivalent La2-xSrxCuO4+y materials are discussed....

  9. Apoptosis-related protein expression in rabbits with blast brain injury following early hyperbaric oxygen therapy

    Shaonian Xu; Jiachuan Liu; Yongming Zhang; Chunlin Wang; Jinbiao Wang; Yanyan Yang; Jian Huo; Wenjiang Sun

    2012-01-01

    We treated detonator-explosion-induced craniocerebral injury in rabbits with hyperbaric oxygen 1-24 hours post-injury. Expression of the apoptosis-regulating protein cytochrome c, the pro-apoptotic protein Bax and the apoptosis marker caspase-3 in the tissues surrounding the area of injury was significantly reduced, while that of the anti-apoptotic protein Bcl-2 was significantly increased. Our findings indicate that the curative effects of early hyperbaric oxygen on cortical cell apoptosis is associated with suppression of cytochrome c release from mitochondria. This mechanism underlies the observed reduction in Bax expression and upregulation of Bcl-2 expression.

  10. Paeonol Protects Memory after Ischemic Stroke via Inhibiting β-Secretase and Apoptosis

    Shan-Yu Su

    2012-01-01

    cells, and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL-positive cells decreased in the paeonol-administered group. Western blotting revealed decreased levels of Bax protein in mitochondria and apoptosis-inducing factor (AIF in cytosol following paeonol treatment. In conclusion, we speculate that paeonol protected memory after ischemic stroke via reducing APP, BACE, and apoptosis. Supression the level of Bax and blocking the release of AIF into cytosol might participate in the anti-apoptosis provided by paeonol.

  11. Ghrelin modulates testicular germ cells apoptosis and proliferation in adult normal rats

    Kheradmand, Arash, E-mail: arashkheradmand@yahoo.com [Department of Clinical Sciences, School of Veterinary Medicine, Lorestan University, P.O. Box: 465, Khorram Abad (Iran, Islamic Republic of); Dezfoulian, Omid [Department of Pathobiology, School of Veterinary Medicine, Lorestan University, Khorram Abad (Iran, Islamic Republic of); Alirezaei, Masoud [Division of Biochemistry, School of Veterinary Medicine, Lorestan University, P.O. Box: 465, Khorram Abad (Iran, Islamic Republic of); Rasoulian, Bahram [Razi Herbal Medicine Research Center, Lorestan University of Medical Sciences, Khorram Abad (Iran, Islamic Republic of)

    2012-03-09

    Highlights: Black-Right-Pointing-Pointer Spermatogenesis is closely associated with the balance between germ cells proliferation and apoptosis. Black-Right-Pointing-Pointer Numerous studies have documented the direct action of ghrelin in the modulation of apoptosis in different cell types. Black-Right-Pointing-Pointer Ghrelin may be considered as a modulator of spermatogenesis in normal adult rats. Black-Right-Pointing-Pointer Ghrelin may be potentially implicated for abnormal spermatogenesis in some testicular germ cell tumors. -- Abstract: Under normal condition in the most mammals, spermatogenesis is closely associated with the balance between germ cells proliferation and apoptosis. The present study was designed to determine the effects of ghrelin treatment on in vivo quality and quantity expression of apoptosis and proliferation specific indices in rat testicular germ cells. Twenty eight adult normal rats were subdivided into equal control and treatment groups. Treatment group received 3 nmol of ghrelin as subcutaneous injection for 30 consecutive days or vehicle to the control animals. The rats from each group (n = 7) were killed on days 10 and 30 and their testes were taken for immunocytochemical evaluation and caspase-3 assay. Immunohistochemical analysis indicated that the accumulations of Bax and PCNA peptides are generally more prominent in spermatocytes and spermatogonia of both groups. Likewise, the mean percentage of immunoreactive spermatocytes against Bax increased (P < 0.01) in the ghrelin-treated group on day 10, while despite of 30% increment in the Bax level of spermatocytes in the treated rats on day 30, however, it was not statistically significant. During the experimental period, only a few spermatogonia represented Bax expression and the changes of Bax immunolabling cells were negligible upon ghrelin treatment. Likewise, there were immunostaining cells against Bcl-2 in each germ cell neither in the control nor in the treated animals. In fact

  12. Caracterización funcional de la molécula proapoptótica Bak en el retículo endoplasmático

    Klee, Martina

    2008-01-01

    Bak y Bax son miembros proapoptóticos de la familia Bcl-2 que presentan una alta redundancia funcional. Ambos homólogos constituyen un punto de control crítico en la vía mitocondrial de apoptosis y están localizados tanto en la mitocondria como en el retículo endoplasmático (RE). Mientras que ya se ha establecido que Bak y Bax mitocondriales son suficientes para la inducción de apoptosis por proteínas BH3-only, todavía se desconoce en gran medida el papel de los homólogos reticulares, aunque ...

  13. Histone deacetylase inhibitors sensitize lung cancer cells to hyperthermia: involvement of Ku70/SirT-1 in thermo-protection.

    Hassan, Mohamed K; Watari, Hidemichi; Salah-Eldin, Alaa-Eldin; Sultan, Ahmed S; Mohamed, Zainab; Fujioka, Yoichiro; Ohba, Yusuke; Sakuragi, Noriaki

    2014-01-01

    This study describes the sensitization mechanism to thermal stress by histone deacetylase inhibitors (HDACIs) in lung cancer cells and shows that Ku70, based on its acetylation status, mediates the protection of lung cancer from hyperthermia (42.5°C, 1-6 hrs). Ku70 regulates apoptosis by sequestering pro-apoptotic Bax. However, its role in thermal stress is not fully understood. The findings showed that, pre-treating lung cancer cells with HDACIs, nicotinamide (NM) or Trichostatin A (TsA) or both significantly enhanced hyperthermia-induced Bax-dependent apoptosis in PC-10 cells. We found that hyperthermia induces SirT-1, Sirtuin, upregulation but not HDAC6 or SirT-3, therefore transfection with dominant negative SirT-1 (Y/H) also eliminated the protection and resulted in more cell death by hyperthermia, in H1299 cells through Bax activation. Hyperthermia alone primed lung cancer cells to apoptosis without prominent death. After hyperthermia Bax was upregulated, Bcl-2 was downregulated, the Bax/Bcl-2 ratio was inversed and Bax/Bcl-2 heterodimer was dissociated. Although hyperthermia did not affect total Ku70 expression level, it stimulated Ku70 deacetylation, which in turn could bind more Bax in the PC-10 cells. These findings suggest an escape mechanism from hyperthermia-induced Bax activation. To verify the role of Ku70 in this protection mechanism, Ku70 was silenced by siRNA. Ku70 silencing significantly sensitized the lung cancer cells to hyperthermia. The Ku70 KD cells underwent cytotoxic G1 arrest and caspase-dependant apoptosis when compared to scrambled transfectants which showed only G2/M cytostatic arrest in the cell lines investigated, suggesting an additional cell cycle-dependent, novel, role of Ku70 in protection from hyperthermia. Taken together, our data show a Ku70-dependent protection mechanism from hyperthermia. Targeting Ku70 and/or its acetylation during hyperthermia may represent a promising therapeutic approach for lung cancer. PMID:24728004

  14. Histone deacetylase inhibitors sensitize lung cancer cells to hyperthermia: involvement of Ku70/SirT-1 in thermo-protection.

    Mohamed K Hassan

    Full Text Available This study describes the sensitization mechanism to thermal stress by histone deacetylase inhibitors (HDACIs in lung cancer cells and shows that Ku70, based on its acetylation status, mediates the protection of lung cancer from hyperthermia (42.5°C, 1-6 hrs. Ku70 regulates apoptosis by sequestering pro-apoptotic Bax. However, its role in thermal stress is not fully understood. The findings showed that, pre-treating lung cancer cells with HDACIs, nicotinamide (NM or Trichostatin A (TsA or both significantly enhanced hyperthermia-induced Bax-dependent apoptosis in PC-10 cells. We found that hyperthermia induces SirT-1, Sirtuin, upregulation but not HDAC6 or SirT-3, therefore transfection with dominant negative SirT-1 (Y/H also eliminated the protection and resulted in more cell death by hyperthermia, in H1299 cells through Bax activation. Hyperthermia alone primed lung cancer cells to apoptosis without prominent death. After hyperthermia Bax was upregulated, Bcl-2 was downregulated, the Bax/Bcl-2 ratio was inversed and Bax/Bcl-2 heterodimer was dissociated. Although hyperthermia did not affect total Ku70 expression level, it stimulated Ku70 deacetylation, which in turn could bind more Bax in the PC-10 cells. These findings suggest an escape mechanism from hyperthermia-induced Bax activation. To verify the role of Ku70 in this protection mechanism, Ku70 was silenced by siRNA. Ku70 silencing significantly sensitized the lung cancer cells to hyperthermia. The Ku70 KD cells underwent cytotoxic G1 arrest and caspase-dependant apoptosis when compared to scrambled transfectants which showed only G2/M cytostatic arrest in the cell lines investigated, suggesting an additional cell cycle-dependent, novel, role of Ku70 in protection from hyperthermia. Taken together, our data show a Ku70-dependent protection mechanism from hyperthermia. Targeting Ku70 and/or its acetylation during hyperthermia may represent a promising therapeutic approach for lung cancer.

  15. The Bcl-2 family member BIM has multiple glaucoma-relevant functions in DBA/2J mice

    Harder, Jeffrey M; Fernandes, Kimberly A; Richard T. Libby

    2012-01-01

    Axonal insult induces retinal ganglion cell (RGC) death through a BAX-dependent process. The pro-apoptotic Bcl-2 family member BIM is known to induce BAX activation. BIM expression increased in RGCs after axonal injury and its induction was dependent on JUN. Partial and complete Bim deficiency delayed RGC death after mechanical optic nerve injury. However, in a mouse model of glaucoma, DBA/2J mice, Bim deficiency did not prevent RGC death in eyes with severe optic nerve degeneration. In a sub...

  16. ERK1/2-dependent phosphorylation of BimEL promotes its rapid dissociation from Mcl-1 and Bcl-xL

    Ewings, Katherine E; Hadfield-Moorhouse, Kathryn; Wiggins, Ceri M; Wickenden, Julie A; Balmanno, Kathryn; Gilley, Rebecca; Degenhardt, Kurt; White, Eileen; Cook, Simon J.

    2007-01-01

    The proapoptotic protein Bim is expressed de novo following withdrawal of serum survival factors. Here, we show that Bim−/− fibroblasts and epithelial cells exhibit reduced cell death following serum withdrawal in comparison with their wild-type counterparts. In viable cells, Bax associates with Bcl-2, Bcl-xL and Mcl-1. Upon serum withdrawal, newly expressed BimEL associates with Bcl-xL and Mcl-1, coinciding with the dissociation of Bax from these proteins. Survival factors can prevent associ...

  17. Ab initio investigation of the surface properties of dispenser B-type and scandate thermionic emission cathodes

    Vlahos, Vasilios; Lee, Yueh-Lin; Booske, John H.; Morgan, Dane; Turek, Ladislav; Kirshner, Mark; Kowalczyk, Richard; Wilsen, Craig

    2009-05-01

    Scandate cathodes (BaxScyOz on W) are important thermionic electron emission materials whose emission mechanism remains unclear. Ab initio modeling is used to investigate the surface properties of both scandate and traditional B-type (Ba-O on W) cathodes. We demonstrate that the Ba-O dipole surface structure believed to be present in active B-type cathodes is not thermodynamically stable, suggesting that a nonequilibrium steady state dominates the active cathode's surface structure. We identify a stable, low work function BaxScyOz surface structure, which may be responsible for some scandate cathode properties and demonstrate that multicomponent surface coatings can lower cathode work functions.

  18. ING1 induces apoptosis through direct effects at the mitochondria

    Bose, P; Thakur, S; Thalappilly, S;

    2013-01-01

    translocates to the mitochondria of primary fibroblasts and established epithelial cell lines in response to apoptosis inducing stimuli, independent of the cellular p53 status. The ability of ING1 to induce apoptosis in various breast cancer cell lines correlates well with its degree of translocation to the...... mitochondria after UV treatment. Endogenous ING1 protein specifically interacts with the pro-apoptotic BCL2 family member BAX, and colocalizes with BAX in a UV-inducible manner. Ectopic expression of a mitochondria-targeted ING1 construct is more proficient in inducing apoptosis than the wild type ING1 protein...

  19. Disease: H00328 [KEGG MEDICUS

    Full Text Available ial use of the bacteria for bioterrorism has focused interest on it. B. anthracis causes symptoms ranging from cutaneous anthrax... that resolves spontaneously to gastrointestinal and inhalational anthrax...ar bat bai bax] Bacillus cereus [GN:bal] anthrax toxin [KO:K11029 K11030 K08645] pore-forming toxin [KO:K110

  20. Upregulation of p‑Akt by glial cell line‑derived neurotrophic factor ameliorates cell apoptosis in the hippocampus of rats with streptozotocin‑induced diabetic encephalopathy.

    Cui, Weigang; Zhang, Yinghua; Lu, Derong; Ren, Mingxin; Yuan, Guoyan

    2016-01-01

    The loss of neurotrophic factor support has been shown to contribute to the development of the central nervous system. Glial cell line‑derived neurotrophic factor (GDNF), a potent neurotrophic factor, is closely associated with apoptosis and exerts neuroprotective effects on numerous populations of cells. However, the underlying mechanisms of these protective effects remain unknown. In the present study, a significant increase in Bax levels and DNA fragmentation was observed in the hippocampus obtained from the brains of diabetic rats 60 days after diabetes had been induced. The apoptotic changes were correlated with the loss of GDNF/Akt signaling. GDNF administration was found to reverse the diabetes‑induced Bax and DNA fragmentation changes. This was associated with an improvement in the level of p‑Akt/Akt. In addition, combination of GDNF with a specific inhibitor of the phosphoinositide 3‑kinase (PI3K)/Akt pathway, Wortmannin, significantly abrogated the effects of GDNF on the levels of p‑Akt/Akt, Bax and DNA fragmentation. However, a p38 mitogen‑activated proten kinase (MAPK) inhibitor, SB203580, had no effect on the expression of p‑Akt/Akt, Bax or DNA fragmentation. These results demonstrate the pivotal role of GDNF as well as the PI3K/Akt pathway, but not the MAPK pathway, in the prevention of diabetes‑induced neuronal apoptosis in the hippocampus. PMID:26549420

  1. Dicty_cDB: Contig-U15582-1 [Dicty_cDB

    Full Text Available pttftrkvaggvkttttgsslp tnntsttttttttaakklpngckisiknsnlltstgltdlddiigggipigsilmieedi nssyymfllkyflaegvlqqqgvffssligidpfeilnklpari... nssyymfllkyflaegvlqqqgvffssligidpfeilnklparitkeeeieadkndttnn intnintnnnnkqptdelk... ) 030630KAWC018590HT (KAWC) Actinidia chinensis - a... 48 1.0 1 ( AM180355 ) Clostridium diff...) Bax-responsive genes for drug target identificati... 40 1.5 6 ( AX537058 ) Sequence 659 from Patent WO0206... 3 Chromosome length 6358359 Start point 1166794 End point 1168269 Strand (PLUS/MINUS) PLUS Number of clones

  2. SwissProt search result: AK104199 [KOME

    Full Text Available AK104199 006-304-A08 (Q8BJZ3) Transmembrane BAX inhibitor motif containing protein 1 (RECS1 prot ... ein) (Responsive to centrifugal force ... and shear stress gene 1 protein) TMBI1_MOUSE 3e-11 ...

  3. SwissProt search result: AK104366 [KOME

    Full Text Available AK104366 001-035-D07 (Q8BJZ3) Transmembrane BAX inhibitor motif containing protein 1 (RECS1 prot ... ein) (Responsive to centrifugal force ... and shear stress gene 1 protein) TMBI1_MOUSE 3e-11 ...

  4. SwissProt search result: AK107547 [KOME

    Full Text Available AK107547 002-130-A05 (Q8BJZ3) Transmembrane BAX inhibitor motif containing protein 1 (RECS1 prot ... ein) (Responsive to centrifugal force ... and shear stress gene 1 protein) TMBI1_MOUSE 1e-14 ...

  5. Effect of digan oral liquid on expression of cell cycle progression and apoptosis-related genes in mice with radiation lesions

    Objective: To study the effect of Digan oral liquid (DGOL) on expression of cell cycle progression (cyclinD1)- and apoptosis(bcl-2 and bax)- related genes in bone marrow of mice with radiation lesions. Methods: After the radiation lesion model (given 6.0 Gy X-rays once by a linear accelerator) was established, the animals were fed with 0.2ml/10 g of either small dose (100%) or large dose (200%) DGOL twice a day for 8 days. On the ninth day cyclinD1, bcl-2 and bax mRNA expression levels in marrow MNC of model mice were measured by in situ hybridization. Results: In the DGOL group, the bax mRNA expression was significantly decreased (P1 and bcl-2 mRNA were significantly increased (P1 and bcl-2 mRNA expression, and inhibitory effect on bax mRNA expression. In this way, it prevents the blood cells from injury, promotes immunity, and protects the organism. This result provides a theoretical basis for clinical application of Digan oral liquid

  6. Potential mechanisms involved in resistant phenotype of MCF-7 breast carcinoma cells to ionizing radiation induced apoptosis

    Wang Yanling [Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730000 (China); Key Laboratory of Heavy Ion Radiation Medicine of Gansu Province, Lanzhou 730000 (China); Graduate School of Chinese Academy of Sciences, Beijing 100039 (China); Zhang Hong [Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730000 (China); Key Laboratory of Heavy Ion Radiation Medicine of Gansu Province, Lanzhou 730000 (China)], E-mail: zhangh@impcas.ac.cn; Li Ning [Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730000 (China); Key Laboratory of Heavy Ion Radiation Medicine of Gansu Province, Lanzhou 730000 (China); Graduate School of Chinese Academy of Sciences, Beijing 100039 (China); Wang Xiaohu [Department of Radiotherapy, Gansu Tumor Hospital, Lanzhou 730050 (China); Hao Jifang [Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730000 (China); Key Laboratory of Heavy Ion Radiation Medicine of Gansu Province, Lanzhou 730000 (China); Zhao Weiping [Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730000 (China); Key Laboratory of Heavy Ion Radiation Medicine of Gansu Province, Lanzhou 730000 (China); Graduate School of Chinese Academy of Sciences, Beijing 100039 (China)

    2009-03-15

    In the present study, we investigated the mechanisms of apoptosis resistance and the roles of the phosphorylation of BRCA1, p21, the Bax/Bcl-2 protein ratio and cell cycle arrest in IR-induced apoptosis in MCF-7 cells. X-irradiation, in particular at low dose (1 Gy), but not carbon ion irradiation, had a significant antiproliferative effect on the growth of MCF-7 cells. 1 Gy X-irradiation resulted in G1 and G2 phase arrest, but 4 Gy induced a significant G1 block. In contrast, carbon ion irradiation resulted in a significant accumulation in the G2 phase. Concomitant with the phosphorylation of H2AX induced by DNA damage, carbon ion irradiation resulted in an approximately 1.9-2.8-fold increase in the phosphorylation of BRCA1 on serine residue 1524, significantly greater than that detected for X-irradiation. Carbon ion irradiation caused a dramatic increase in p21 expression and drastic decrease in Bax expression compared with X-irradiation. The data implicated that phosphorylation of BRCA1 on serine residue 1524 might, at least partially, induce p21 expression but repress Bax expression. Together, our results suggested that the phosphorylation of BRCA1 at Ser-1524 might contribute to the G2 phase arrest and might be an upstream signal involved in preventing apoptosis signal via upregulation of p21 and downregulation of the Bax/Bcl-2 ratio.

  7. Potential mechanisms involved in resistant phenotype of MCF-7 breast carcinoma cells to ionizing radiation induced apoptosis

    Wang, Yan-ling; Zhang, Hong; Li, Ning; Wang, Xiao-hu; Hao, Ji-fang; Zhao, Wei-ping

    2009-03-01

    In the present study, we investigated the mechanisms of apoptosis resistance and the roles of the phosphorylation of BRCA1, p21, the Bax/Bcl-2 protein ratio and cell cycle arrest in IR-induced apoptosis in MCF-7 cells. X-irradiation, in particular at low dose (1 Gy), but not carbon ion irradiation, had a significant antiproliferative effect on the growth of MCF-7 cells. 1 Gy X-irradiation resulted in G1 and G2 phase arrest, but 4 Gy induced a significant G1 block. In contrast, carbon ion irradiation resulted in a significant accumulation in the G2 phase. Concomitant with the phosphorylation of H2AX induced by DNA damage, carbon ion irradiation resulted in an approximately 1.9-2.8-fold increase in the phosphorylation of BRCA1 on serine residue 1524, significantly greater than that detected for X-irradiation. Carbon ion irradiation caused a dramatic increase in p21 expression and drastic decrease in Bax expression compared with X-irradiation. The data implicated that phosphorylation of BRCA1 on serine residue 1524 might, at least partially, induce p21 expression but repress Bax expression. Together, our results suggested that the phosphorylation of BRCA1 at Ser-1524 might contribute to the G2 phase arrest and might be an upstream signal involved in preventing apoptosis signal via upregulation of p21 and downregulation of the Bax/Bcl-2 ratio.

  8. Effects of Oxymatrine on the Apoptosis of Human Esophageal Carcinoma Eca109 Cell Line and Its Mechanism

    Yi JIN; Jianli HU; Qiong WANG; Zhenyu LI; Yeshan CHEN

    2008-01-01

    The effects of Oxymatrine (Oxy) on the proliferation and apoptosis of human esophageal carcinoma Eca109 cell line and the mechanism were investigated. The human esophageal carcinoma Eca109 cells were cultured in vitro. The Oxy-induced apoptosis of Eca109 cells was assayed by using flow cytometry. The expressions of p-ERK1/2, Cyclin D1, p21waf/cip1, Bax and Bcl-2 were detected by Western blot. Flow cytometry revealed that Oxy could induce the apoptosis of Eca109 cells. Western blot showed that Oxy of different concentrations suppressed the expressions of p-ERK1/2, cyclin D1 and Bc1-2, but up-regulated the expression of p21waf/cip1 and Bax, and the ratio of Bax/Bcl-2 was increased It was suggested the Oxy could induce the apoptosis of Eca109-cells, which might be related to the upregulation of p21waf/cip1 and the downregulation of p-ERK1/2 Cyclin D1 and p21waf/cip1. The possible pathway may be related to Bcl-2/Bax.

  9. Dicty_cDB: Contig-U15988-1 [Dicty_cDB

    Full Text Available Sequence 862 from patent US 7135558. 44 4.1 1 ( DL173691 ) Methods for Identifying the Target of a Compound ...... 44 4.1 1 ( DL173298 ) Methods for Identifying the Target of a Compound ... 44 4.1 1 ( DL131093 ) Bax-res

  10. ICT in ELT: How Did We Get Here and Where Are We Going?

    Dudeney, Gavin; Hockly, Nicky

    2012-01-01

    This article looks at how specific developments in information and communication technologies (ICT) have impacted on ELT over the past three decades. Of particular interest is the effect on classroom practice, and on the types of materials available for teaching and learning. We take as our starting point Mark Warschauer's and Stephen Bax's…

  11. Deficient Fas expression by CD4+ CCR5+ T cells in multiple sclerosis

    Julià, Eva; Montalban, Xavier; Al-Zayat, Hammad; Issazadeh-Navikas, Shohreh; Goertsches, Robert; Martin, Roland; Comabella, Manuel

    OBJECTIVE: To evaluate whether T cells expressing CCR5 and CXCR3 from multiple sclerosis (MS) patients are more resistant to apoptosis. METHODS: Expression of CD69, TNF-R1, Fas, FasL, bcl-2, and bax was investigated in 41 MS patients and 12 healthy controls by flow cytometry in CD4+ and CD8+ T ce...

  12. Medium-intensity acute exhaustive exercise induces neural cell apoptosis in the rat hippocampus

    Shanni Li; Jin Liu; Hengmei Yan

    2013-01-01

    The present study assessed the influence of medium-intensity (treadmill at a speed of 19.3 m/min until exhaustion) and high-intensity (treadmill at a speed of 26.8 m/min until exhaustion) acute exhaustive exercise on rat hippocampal neural cell apoptosis. TUNEL staining showed significantly increased neural cell apoptosis in the hippocampal CA1 region of rats after medium- and high-intensity acute exhaustive exercise, particularly the medium-intensity acute exhaustive exercise, when compared with the control. Immunohistochemistry showed significantly increased expression of the antiapoptotic protein Bcl-2 and the proapoptotic protein Bax in the hippocampal CA1 region of rats after medium- and high-intensity acute exhaustive exercise. Additionally, the ratio of Bax to Bcl-2 increased in both exercise groups. In particular, the medium-intensity acute exhaustive exercise group had significantly higher Bax and Bcl-2 protein expression and a higher Bax/Bcl-2 ratio. These findings indicate that acute exhaustive exercise of different intensities can induce neural cell apoptosis in the hippocampus, and that medium-intensity acute exhaustive exercise results in greater damage when compared with high-intensity exercise.

  13. Medium-intensity acute exhaustive exercise induces neural cell apoptosis in the rat hippocampus.

    Li, Shanni; Liu, Jin; Yan, Hengmei

    2013-01-15

    The present study assessed the influence of medium-intensity (treadmill at a speed of 19.3 m/min until exhaustion) and high-intensity (treadmill at a speed of 26.8 m/min until exhaustion) acute exhaustive exercise on rat hippocampal neural cell apoptosis. TUNEL staining showed significantly increased neural cell apoptosis in the hippocampal CA1 region of rats after medium- and high-intensity acute exhaustive exercise, particularly the medium-intensity acute exhaustive exercise, when compared with the control. Immunohistochemistry showed significantly increased expression of the antiapoptotic protein Bcl-2 and the proapoptotic protein Bax in the hippocampal CA1 region of rats after medium- and high-intensity acute exhaustive exercise. Additionally, the ratio of Bax to Bcl-2 increased in both exercise groups. In particular, the medium-intensity acute exhaustive exercise group had significantly higher Bax and Bcl-2 protein expression and a higher Bax/Bcl-2 ratio. These findings indicate that acute exhaustive exercise of different intensities can induce neural cell apoptosis in the hippocampus, and that medium-intensity acute exhaustive exercise results in greater damage when compared with high-intensity exercise. PMID:25206482

  14. Metabolism and Endocrinology

    2011-01-01

    2011183 Effect of pioglitazone on the expressions of Bax and Bcl-2 protein in mouse osteoblasts. LI Jun-yan (李俊岩) ,et al. Dept Endocrinol,Heji Hosp, Changzhi Med Coll,Changzhi 046000. Chin J Endocrinol Metab 2011; 27(2):118-121.

  15. Paclitaxel induces apoptosis in human gastric carcinoma cells

    Hai-Bo Zhou; Ju-Ren Zhu

    2003-01-01

    AIM: To investigate the apoptosis in gastric cancer cells induced by paclitaxel, and the relation between this apoptosis and expression of Bcl-2 and Bax.METHODS: In in vitro experiments, MTT assay was used to determine the cell growth inhibitory rate. Transmission electron microscope and TUNEL staining method were used to quantitatively and qualitively detect the apoptosis status of gastric cancer cell line SGC-7901 before and after the paditaxel treatment. Immunohistochemical staining was used to detect the expression of apoptosis-regulated gene Bcl-2and Bax.RESULTS: Paclitaxel inhibited the growth of gastric cancer cell line SGC-7901 in a dose-and time-dependent manner.Paclitaxel induced SGC-7901 cells to undergo apoptosis with typically apoptotic characteristics, including morphological changes of chromatin condensation, chromatin crescent formation, nucleus fragmentation and apoptotic body formation. Paclitaxel could reduce the expression of apoptosis-regulated gene Bcl-2, and improve the expression of apoptosis-regulated gene Bax.CONCLUSION: Paclitaxel is able to induce the apoptosis in gastric cancer. This apoptosis may be mediated by downexpression of apoptosis-regulated gene Bcl-2 and upexpression of apoptosis-regulated gene Bax.

  16. Combinatorial chemopreventive effect of butyric acid, nicotinamide and calcium glucarate against the 7,12-dimethylbenz(a)anthracene induced mouse skin tumorigenesis attained by enhancing the induction of intrinsic apoptotic events.

    Tiwari, Prakash; Sahay, Satya; Pandey, Manuraj; Qadri, Syed S Y H; Gupta, Krishna P

    2015-01-25

    We explored the basis of the combinatorial chemopreventive effect of butyric acid (BA), nicotinamide (NA) and calcium glucarate (CAG) on mouse skin exposed to 7,12-dimethylbenz(a)anthracene (DMBA). We studied the effects of topical application of DMBA in the presence or absence of BA, NA and CAG on the regulators of apoptosis. DMBA treatment suppressed Bax, Bax/Bcl-2 ratio, release of cyt c, Apaf1, caspase-9, -3 mediated apoptosis. Downregulation of p21 and upregulation of Bcl-2, mut p53 were also observed in only DMBA treated mice. Simultaneous application of BA, NA and CAG induced a mitochondria-mediated apoptosis, characterized by a rise in the Bax, Bax/Bcl-2 ratio, release of cyt c, upregulation of Apaf1 with down-stream activation of caspase-9, -3. Furthermore treatment with BA, NA and CAG demonstrated an upregulation of p21 and downregulation of Bcl-2, mut p53. But this effect was enhanced in the presence of all the three compounds together in combination. Chemoprevention by a combination of BA, NA and CAG by inducing the apoptosis, the natural cell death, suggest the importance of the potential combinational strategies capable of preventing skin tumor development. PMID:25478867

  17. Communicative Language Teaching: Unity within Diversity

    Hiep, Pham Hoa

    2007-01-01

    Recent articles in the "ELT Journal" offer interesting debates on CLT. On one side, Bax (2003) proposes that CLT should be abandoned since the methodology fails to take into account the context of language teaching. On the other side, Liao (2004) suggests that CLT is best. However, within the broad theoretical position on which CLT is based,…

  18. Vitamin E-deficiency did not exacerbate partial skin reactions in mice locally irradiated with X-rays

    We previously showed that free radicals and oxidative stress are involved in radiation-induced skin reactions. Since vitamin E (VE) is a particularly important lipophilic antioxidant, VE-deficient mice were used to examine its effects on radiation-induced skin damage. The VE content of the skin was reduced to one fourth of levels of normal mice. Neither the time of onset nor the extent of the reactions quantified with a scoring system differed between normal and VE-deficient mice after local X-irradiation (50 Gy). Similarly, there was no difference in the levels of the ascorbyl radical between the groups, although they were higher in irradiated skin than non-irradiated skin. X-irradiation increased the amount of Bax protein in the skin of normal mice both in the latent and acute inflammatory stages, time- and dose-dependently. The increase was associated with an increase in cytochrome c in the cytosolic fraction, indicating that apoptosis was also promoted by the irradiation. The increase in Bax protein correlated well with the thickness of the skin. Although a deficiency in VE should lower resistance to free radicals in the mitochondrial membrane and thus enhance radiation-induced Bax expression and apoptosis, it actually attenuated the increase in Bax protein caused by irradiation. (author)

  19. Inhibition of NF-κB enhanced X-ray irradiation induced apoptosis in human non-Hodgkin lymphoma

    Objective: To investigate the role of NF-κB in radiation-induced apoptosis to human non-Hodgkin lymphoma (NHL) cells and the mechanism involved. Methods: Three human non-Hodgkin lymphoma cell lines, Namalwa, Ramos, and Raji cells were divided into control, IR and IR + QNZ (10 nmol/L) groups, respectively.Annexin-V kit was used to determine cell apoptosis. Protein expression levels of Survivin, Bax, Bcl-2 and cleaved Caspase-3 were evaluated by Western blot. Survivin mRNA was quantified by real-time PCR. Results: Inhibition of NF-κB by QNZ pretreatment significantly enhanced X-ray induced apoptosis in human NHL cells in a dose-dependent manner (t=2.93-12.52, P<0.05). At the same time, QNZ significantly reversed the expression levels of Survivin protein and mRNA that were up-regulated by radiation (t=3.29-16.72, P<0.05). QNZ also increased the levels of Caspase-3 and pro-apoptotic protein Bax, but reduced anti-apoptotic protein Bcl-2 expression level and hence the ratio of Bcl-2/Bax. Conclusions: Inhibition of NF-kB could enhance radiation-induced cell apoptosis in human NHL cells through down-regulating Survivin expression and decreasing Bcl-2/Bax ratio. (authors)

  20. Experimental Study on the Preventive Mechanism of Salviae Miltiorrhizae Against Atherosclerosis in Rabbits Models

    李树生; 万磊

    2004-01-01

    Summary: The preventive mechanism of salviae miltiorrhizae (SM) against experimental atherosclerosis (AS) in rabbits models was investigated. The experimental AS rabbit models were reproduced by feeding the high cholesterol diet. The changes of atherosclerotic plaques in normal group, model group and SM treated group were observed. The levels of serum TG, TC, HDL-C and LDL-C were determined. The immunohistochemistry was used to detect the expression of Bcl-2,Bax and IL-6 proteins in atherosclerotic plaques. The results showed that the level of serum TG in SM treated group was significantly lower than in model group (P<0.01). Immunohistochemistry revealed that the expression of Bcl-2, Bax ano IL-6 in model group was significantly higher than in normal group.In the SM group, the expression of Bcl-2 protein was up-regulated and that of Bax was down-regulated. It was suggested that SM could inhibit formation of AS in experimental rabbits. To decrease the expression of Bax and increase the expression of Bcl-2 protein may be one of the mechanisms of SM against atherosclerosis.

  1. Apoptosis of human pancreatic cancer cells induced by Triptolide

    Guo-Xiong Zhou; Xiao-Ling Ding; Jie-Fei Huang; Hong Zhang; Sheng-Bao Wu; Jian-Ping Cheng; Qun Wei

    2008-01-01

    AIM:To investigate apoptosis in human pancreatic cancer ceils induced by Triptolide (TL),and the relationship between this apoptosis and expression of caspase-3' bcl-2 and bax.METHODS:Human pancreatic cancer cell line SW1990 was cultured in DIEM media for this study.MTT assay was used to determine the cell growth inhibitory rate in vitro.Flow cytometry and TUNEL assay were used to detect the apoptosis of human pancreatic cancer cells before and after TL treatment.RT-PCR was used to detect the expression of apoptosis-associated gene caspase-3' bcl-2 and bax.RESULTS:TL inhibited the growth of human pancreatic cancer cells in a dose-and time-dependent manner.TL induced human pancreatic cancer cells to undergo apoptosis with typically apoptotic characteristics.TUNEL assay showed that after the treatment of human pancreatic cancer cells with 40 ng/mL TL for 12 h and 24 h,the apoptotic rates of human pancreatic cancer cells increased significantly.RT-PCR demonstrated that caspase-3 and bax were significantly up-regulated in SW1990 cells treated with TL while bcl-2 mRNA was not.CONCLUSION:TL is able to induce the apoptosis in human pancreatic cancer cells.This apoptosis may be mediated by up-regulating the expression of apoptosisassociated caspase-3 and bax gene.

  2. Cardiolipin or MTCH2 can serve as tBID receptors during apoptosis.

    Raemy, E; Montessuit, S; Pierredon, S; van Kampen, A H; Vaz, F M; Martinou, J-C

    2016-07-01

    During apoptosis, proapoptotic BAX and BAK trigger mitochondrial outer membrane (MOM) permeabilization by a mechanism that is not yet fully understood. BH3-only proteins such as tBID, together with lipids of the MOM, are thought to play a key role in BAX and BAK activation. In particular, cardiolipin (CL) has been shown to stimulate tBID-induced BAX activation in vitro. However, it is still unclear whether this process also relies on CL in the cell, or whether it is more dependent on MTCH2, a proposed receptor for tBID present in the MOM. To address this issue, we deleted both alleles of cardiolipin synthase in human HCT116 cells by homologous recombination, which resulted in a complete absence of CL. The CL-deficient cells were fully viable in glucose but displayed impaired oxidative phosphorylation and an inability to grow in galactose. Using these cells, we found that CL was not required for either tBID-induced BAX activation, or for apoptosis in response to treatment with TRAIL. Downregulation of MTCH2 in HCT116 cells also failed to prevent recruitment of tBID to mitochondria in apoptotic conditions. However, when both CL and MTCH2 were depleted, a significant reduction in tBID recruitment was observed, suggesting that in HCT116 cells, CL and MTCH2 can have redundant functions in this process. PMID:26794447

  3. Apoptosis of Alveolar Wall Cells in Chronic Obstructive Pulmonary Disease Patients with Pulmonary Emphysema Is Involved in Emphysematous Changes

    Hongmei LIU; Lijun MA; Jizhen WU; Kai WANG; Xianliang CHEN

    2009-01-01

    s of alveolar wall cells, espe-cially apoptosis of type-Ⅱ cells, may take part in the pathogenesis of emphysema. Up-regulation of Bax expression may be responsible for the apoptosis of alveolar wall cells in the COPD patients with pulmonary emphysema.

  4. Genetic dissection of apoptosis and cell cycle control in response of colorectal cancer treated with preoperative radiochemotherapy

    In previous analyses we identified therapy-induced upregulation of the CDK inhibitor p21CIP/WAF-1 and consequently decreased tumor cell proliferation or loss of Bax as adverse factors for survival in rectal cancer treated with radiochemotherapy. Here, we address the individual role of p53 and its transcriptional targets, p21CIP/WAF-1 and Bax, on apoptosis induced by individual components of multimodal anticancer therapy, i.e. 5-fluorouracil (5-FU), ionising γ-radiation (IR) and heat shock/hyperthermia. We analysed tumor samples 66 patients with rectal carcinoma treated by a neoadjuvant approach with radiochemotherapy ± heat shock/hyperthermia for the expression and mutation of p53 and the expression of p21CIP/WAF-1 and Bax. These data were correlated with the tumor response. The functional relevance of p53, p21CIP/WAF-1 and Bax was investigated in isogeneic HCT116 cell mutants treated with 5-FU, IR and heat shock. Rectal carcinoma patients who received an optimal heat shock treatment showed a response that correlated well with Bax expression (p = 0.018). Local tumor response in the whole cohort was linked to expression of p21CIP/WAF-1 (p < 0.05), but not p53 expression or mutation. This dichotomy of p53 pathway components regulating response to therapy was confirmed in vitro. In isogeneic HCT116 cell mutants, loss of Bax but not p53 or p21CIP/WAF-1 resulted in resistance against heat shock. In contrast, loss of p21CIP/WAF-1 or, to a lesser extent, p53 sensitized predominantly for 5-FU and IR. These data establish a different impact of p53 pathway components on treatment responses. While chemotherapy and IR depend primarily on cell cycle control and p21, heat shock depends primarily on Bax. In contrast, p53 status poorly correlates with response. These analyses therefore provide a rational approach for dissecting the mode of action of single treatment modalities that may be employed to circumvent clinically relevant resistance mechanisms in rectal cancer

  5. Transcription of five p53- and Stat-3-Inducible genes after ionizing radiation

    Ionizing radiation (IR) produces temporal- and dose-dependent changes in multiple gene mRNA targets that are potential biomarkers of radiation dose. We confirmed IR-induced changes in expression of gadd45a, ddb-2, and cdkn1a downstream transcripts of p53 by quantitative reverse transcription-polymerase chain reaction (QRT-PCR) assay in total RNA samples from the whole blood of radiotherapy patients undergoing total-body irradiation [Amundson, S.A., Grace, M.B., McLeland, C.B., Epperly, M.W., Yeager, A., Zhan, Q., Greenberger, J.S., Fornace Jr., A.J., 2004. Human in vivo radiation-induced biomarkers: gene expression changes in radiotherapy patients. Cancer Res. 64, 6368-6371.]. We now confirm dose-dependent up-regulation of bax in addition to these p53-dependent transcripts, and bcl-2, a downstream transcript of Stat-3, in ex vivo irradiated blood samples from healthy unrelated volunteers. Together these biomarkers represent pathways involved in growth arrest, DNA damage, and apoptosis. The objectives of this study were to (1) investigate the relationship between baseline mRNA expression levels, and (2) define expression patterns in response to IR in a large cohort (n=20). Whole-blood samples were irradiated ex vivo to measure gene expression in samples from (i) three healthy donors over a broad dose range (0, 0.25, 0.50, 0.75, 1, 2, and 3 Gy), and (ii) 20 healthy donors at two doses, 0.25 and 2.5 Gy. Expression level variance (σ2) of baseline values (0 Gy) showed negligible inter-individual variation with all values ≤1.0. σ2values=0.50bax, 0.25 bcl-2, 0.73 gadd45a, 0.66 cdkn1a, and 1.0 ddb-2. Meaningful IR dose-responses were observed for bax, gadd45a, and ddb-2 profiles and the ratio of bax:bcl-2 mRNA expression over a broad dose range. QRT-PCR studies were extended in the lower dose range (0, 0.1, 0.5, 0.75, and 1 Gy). Results showed that bax:bcl-2 ratio initially favors bax expression at doses of <1Gy, with IR-induced dose responses observed between 1 and 3

  6. Effect of fetal hypothyroidism on tolerance to ischemia-reperfusion injury in aged male rats: Role of nitric oxide.

    Jeddi, Sajad; Zaman, Jalal; Ghasemi, Asghar

    2016-05-01

    Aging is associated with increased prevalence of cardiovascular disease. Thyroid hormone deficiency during fetal life decreases myocardial tolerance to ischemia-reperfusion (IR) injury in later life. The long-term effects of fetal hypothyroidism (FH) on response to IR injury in aged rats have not been well documented. The aim of this study was therefore to compare the effect of FH on tolerance to IR injury in young and aged male rats and to determine contribution of iNOS (inducible nitric oxide synthase), Bax, and Bcl-2. Pregnant female rats were divided into two groups: The FH group received water containing 0.025% 6-propyl-2-thiouracil during gestation and the controls consumed tap water. Isolated perfused hearts from young (3 months) and aged (12 months) rats were subjected to IR. Hemodynamic parameters, infarct size, and heart NOx (nitrite+nitrate) levels were measured; in addition, mRNA expression of iNOS, Bax, and Bcl-2 and their protein levels in heart were measured. Recovery of post-ischemic LVDP and ±dp/dt were lower and infarct sizes were higher than controls in aged FH rats (68.38 ± 6.7% vs. 50.5 ± 1.7%; P Bcl-2 was lower in both the young (350 and 240% for iNOS and Bax, respectively and 51% for Bcl-2) and aged rats (504 and 567% for iNOS and Bax, respectively and 67% for Bcl-2). Compared to controls, in FH rats protein levels of iNOS (37% for young and 45% for aged rats) and Bax (94% for young and 118% for aged rats) were higher while for Bcl-2 (36% for young and 62% for aged rats) were lower. After IR, in FH rats, aminoguanidine, a selective iNOS inhibitor, decreased mRNA expression of iNOS and Bax and increased expression of Bcl-2 in both young (65% and 58% for iNOS and Bax, respectively and 152% for Bcl-2) and aged rats (76% and 64% for iNOS and Bax, respectively and 222% for Bcl-2). In addition, in the heart of FH rats, aminoguanidine decreased protein levels of iNOS (47% for young and 60% for aged rats) and Bax (57% for young and 80% for

  7. Induction of Apoptosis and expression of Apoptosis-related gene products in response to radiation in murine tumors

    To analyze the involvement of apoptosis regulatory genes p53, p21waf1/cip1, bax and bcl-2 in induction of apoptosis by radiation in murine tumors. The radiation-sensitive ovarian carcinoma OCa-I and the radiation-resistant hepatocarcinoma HCa-I were used. Tumors, 8mm in diameter, were irradiated with 25Gy and at various times after irradiation, ranging from 1 to 48 h, were analyzed histologically for apoptosis and by western blot for alterations in the expression of these genes. The p53 status of the tumors were determined by the polymerase chain reaction-single strand conformation polymorphism assay. Both tumors were positive for wild-type p53. Radiation induced apoptosis in OCa-I but not in HCa-I. Apoptosis developed rapidly, peaked at 2 h after irradiation and returned to almost the background level at 48 h. In OCa-I radiation upregulated the expression of p53, p21waf1/cip1, and the bcl-2/bax ratio was decreased. In HCa-I radiation increased the expression of both p53 and p21waf1/cip1, although the increase of the latter was small. The bcl-2/bax ratio was greatly increased. In general the observed changes occurred within a few hours after irradiation, and either preceded or coincided with development of apoptosis. The development of apoptosis required upregulation of both p53 and p21waf1/cip1 as well as a decrease in bcl-2/bax ratio. In contrast, an increase in bcl-2/bax ratio prevented apoptosis in the presence of upregulated p53 and p21waf1/cip1. These findings identified the involvement of multiple oncogenes in apoptosis regulation in vivo and demonstrate the complexity that may be associated with the use of a single oncogene assessment for predicting the outcome of cancer therapy with cytotoxic agents. (author)

  8. Change in expression of apoptosis genes after hyperthermia, chemotherapy and radiotherapy in human colon cancer transplanted into nude mice

    2007-01-01

    AIM: To investigate the change in expression of p53, Bcl-2, and Bax genes in human colon cancer cells transplanted into nude mice after hyperthermia,chemotherapy, radiotherapy, thermochemotherapy,thermoradiotherapy and thermochemoradiotherapy.METHODS: Human colon cancer cell line (HT29)was transplanted into the hind limbs of nude mice.Under laboratory simulated conditions of hyperthermia (43℃, 60 min), the actual radiation doses and doses of mitomycin C (MMC) were calculated in reference to the clinical radiotherapy for human rectal cancer and chemotherapy prescription for colon cancer. The mice were divided into 6 groups according to the treatment approaches: hyperthermia, chemotherapy,radiotherapy, thermochemotherapy, thermoradiotherapy,and thermochemoradiotherapy. The mice were sacrificed at different time points and the tumor tissue was taken for further procedures. The morphologic changes in membrane, cytoplasm and nuclei of tumor cells of p53, Bcl-2, and Bax after treatment, were observed by immunohistochemistry staining.RESULTS: All of the six treatment modalities downregulated the expression of p53, Bcl-2 and up-regulated the expression of Bax at different levels. The combined therapy of hyperthermia, with chemotherapy, and/or irradiation showed a greater effect on down-regulating the expression of p53 (0.208 ± 0.009 vs 0.155 ± 0.0115,P < 0.01) and Bcl-2 (0.086 ± 0.010 vs 0.026 ± 0.0170,P < 0.01) and up-regulating Bax expression (0.091 ±0.0013 vs 0.207 ±0.027, P < 0.01) compared with any single therapy.CONCLUSION: Hyperthermia enhances the effect of radio- and chemotherapy on tumors by changing the expression of apoptosis genes, such as p53, Bcl-2 and Bax.

  9. Protective Effect and Mechanism of TGF - β1 on Intestinal Mucosa against IR Injury in Rats

    SUN Peichun; WU Wanqing; WU Gang

    2008-01-01

    Objective To investigate the protective effect and mechanism of TGF - 1 on intestinal mucosa against ischemia-reperfusion (I/R) injury, so as to provide new methods of clinical prevention from and dealing with intestine I/R injury and gut - derived multiple organ dysfunction syndrome(MODS). Methods 30 SD rats were randomly divided into 3 groups ( n = 10 each), A: namely sham - operation group ( group A) : superior mesenteric artery ( SMA ) was only isolated but not blocked; B: control group (group B), ischemia for 60 minutes, then reperfusion for 120 minutes, and administered with normal saline solution (NS) 0. 5ml through SMA 10 minutes before SMA blockage; C: experimental group (group C) : animals underwent the same procedure with group B except for taking TGF-β1 instead of NS. The contents of intestinal mucosa Chiu's sore, bacteria translocation, intestinal mucosa cellular apoptosis index, expression of bax mRNA and bcl - 2 mRNA and protein were measured. Results Compared with group A, the results from group B are as follows, the Chiu's score significantly increased, bacteria translocation increased, intestinal mucosa cellular apeptosis rate significantly increased, level of bax mRNA and Bax protein significantly increased, while level of intestinal mucosal bcl -2 mRNA and Bcl -2 protein significantly decreased. All the changes in group B were reversed significantly in group C. Conclusion TGF - 1 exerts a protective effect on the gut barrier against intestine I/R injury, perhaps via reversing injury - induced downregulation in bcl2 transcription and upregulation in bax transcription, with the result in modulating bcl -2 and bax protein expression and downregulation of the susceptibility to apoptosis of intestinal epithelial cells.

  10. Quantification of protein copy number in single mitochondria: The Bcl-2 family proteins.

    Chen, Chaoxiang; Zhang, Xiang; Zhang, Shuyue; Zhu, Shaobin; Xu, Jingyi; Zheng, Yan; Han, Jinyan; Zeng, Jin-Zhang; Yan, Xiaomei

    2015-12-15

    Bcl-2 family proteins, represented by antiapoptotic protein Bcl-2 and proapoptotic protein Bax, are key regulators of mitochondria-mediated apoptosis pathway. To build a quantitative model of how Bcl-2 family protein interactions control mitochondrial outer membrane permeabilization and subsequent cytochrome c release, it is essential to know the number of proteins in individual mitochondria. Here, we report an effective method to quantify the copy number and distribution of proteins in single mitochondria via immunofluorescent labeling and sensitive detection by a laboratory-built high sensitivity flow cytometer (HSFCM). Mitochondria isolated from HeLa cells were stained with Alexa Fluor 488 (AF488)-labeled monoclonal antibodies specifically targeting Bcl-2 or Bax and with nucleic acid dye. A series of fluorescent nanospheres with fluorescence intensity calibrated in the unit of molecules of equivalent soluble fluorochrome (MESF)-AF488 were used to construct a calibration curve for converting the immunofluorescence of a single mitochondrion to the number of antibodies bound to it and then to the number of proteins per mitochondrion. Under the normal condition, the measured mean copy numbers were 1300 and 220 per mitochondrion for Bcl-2 and Bax, respectively. A significant variation in protein copy number was identified, which ranged from 130 to 6000 (2.5-97.5%) for Bcl-2 and from 65 to 700 (2.5-97.5%) for Bax, respectively. We observed an approximately 4.4 fold increase of Bax copy number per mitochondrion upon 9h of apoptosis stimulation while the abundance of Bcl-2 remained almost unchanged. To the best of our knowledge, this is the first report of Bcl-2 family protein copy number and variance in single mitochondria. Collectively, we demonstrate that the HSFCM-based immunoassay provides a rapid and sensitive method for determining protein copy number distribution in single mitochondria. PMID:26176207

  11. Astragaloside IV, a novel antioxidant, prevents glucose-induced podocyte apoptosis in vitro and in vivo.

    Dingkun Gui

    Full Text Available Glucose-induced reactive oxygen species (ROS production initiates podocyte apoptosis, which represents a novel early mechanism leading to diabetic nephropathy (DN. Here, we tested the hypothesis that Astragaloside IV(AS-IV exerts antioxidant and antiapoptotic effects on podocytes under diabetic conditions. Apoptosis, albuminuria, ROS generation, caspase-3 activity and cleavage, as well as Bax and Bcl-2 mRNA and protein expression were measured in vitro and in vivo. Cultured podocytes were exposed to high glucose (HG with 50, 100 and 200 µg/ml of AS-IV for 24 h. AS-IV significantly attenuated HG-induced podocyte apoptosis and ROS production. This antiapoptotic effect was associated with restoration of Bax and Bcl-2 expression, as well as inhibition of caspase-3 activation and overexpression. In streptozotocin (STZ-induced diabetic rats, severe hyperglycemia and albuminuria were developed. Increased apoptosis, Bax expression, caspase-3 activity and cleavage while decreased Bcl-2 expression were detected in diabetic rats. However, pretreatment with AS-IV (2.5, 5, 10 mg·kg(-1·d(-1 for 14 weeks ameliorated podocyte apoptosis, caspase-3 activation, renal histopathology, podocyte foot process effacement, albuminuria and oxidative stress. Expression of Bax and Bcl-2 mRNA and protein in kidney cortex was partially restored by AS-IV pretreatment. These findings suggested AS-IV, a novel antioxidant, to prevent Glucose-Induced podocyte apoptosis partly through restoring the balance of Bax and Bcl-2 expression and inhibiting caspase-3 activation.

  12. The effect of arsenic trioxide on human hepatoma cell line BEL-7402 culturedin vitro

    You Lin Yang; Hong Yu Xu; Yuan Yuan Gao; Qiao Li Wu; Guang Qiang Gao

    2000-01-01

    AIM To study the effect of a wide range of concentration of arsenic trioxide on human hepatoma cell lineBEL-7402 and its mechanism.METHODS The BEL-7402 cells were treated with arsenic trioxide (a final concentration of 0.5, 1 and2 μmol/L, respectively) in various durations or for 4 successive days. The cell growth and proliferation wereobserved by cell counting and cell-growth curve. Morphologic changes were studied under electronmicroscopy. Flow cytometry was used to assay cell-DNA distribution and the protein expression of Bcl-2 andBax was detected by immunocytochemical method.RESULTS The cell growth was significantly inhibited by the different concentrations of arsenic trioxide asrevealed by cell counting and cell-growth curve. Arsenic trioxide treatment at 0.5, 1 and 2 μmol/L, resultedin a sub-G1 cell peak. The decreased G0/G1 phase cell and the increased percentage of S phase cell were observed by flow cytometer, suggesting that the inhibiting effect of arsernic trioxide on BEL-7402 cell lay inG0/G1 phase cell. Apoptotis-related morphology, such as intact cell membrane, nucleic condensation,apoptotic body formation, can be seen under the electron microscopy. High protein expression level of Bcl-2and Bax was detected in 1 and 2 μmol/L arsenic trioxide-treated cells, but that of Bax was more significant.Arsenic trioxide treatment at 0.5 μmol/L resulted in higher expression level of Bcl-2 and lower expressionlevel of Bax compared with control (P1<0.01, P2<0.01).CONCLUSION Arsenic trioxide not only inhibited the proliferation but also induced apoptosis of humanhepatoma cell line BEL-7402. The induced-apoptosis effect of 1 and 2 μmol/L arsenic trioxide was relative tothe expression level of Bcl-2 and Bax.

  13. Clostridium butyricum pretreatment attenuates cerebral ischemia/reperfusion injury in mice via anti-oxidation and anti-apoptosis.

    Sun, Jing; Ling, Zongxin; Wang, Fangyan; Chen, Wenqian; Li, Haixiao; Jin, Jiangtao; Zhang, Huiqing; Pang, Mengqi; Yu, Junjie; Liu, Jiaming

    2016-02-01

    Probiotics participate actively in the neuropsychiatric disorders. However, their roles on ischemic stroke remain unclear. This study aims to determine whether Clostridium butyricum (C. butyricum) could attenuate cerebral ischemia/reperfusion (I/R) injury and its possible mechanisms. Male ICR mice were intragastrically pretreated with C. butyricum for 2 successive weeks, and then subjected to cerebral I/R injury induced by the bilateral common carotid artery occlusion (BCCAO) for 20min. After 24h of the reperfusion, neurological deficit scores were evaluated. Histopathological changes of the hippocampus neurons were observed using Hematoxylin and eosin (H&E) and TUNEL staining. Malondialdehyde (MDA) contents and superoxide dismutase (SOD) activities in the brain were detected. The expression of Caspase-3, Bax and Bcl-2 were investigated by Western blot and immunohistochemistry analysis. The butyrate contents in the brain were determined. Our results showed that cerebral I/R injury led to neurological deficit, increased levels of Caspase-3 and Bax and decreased Bcl-2/Bax ratio. C. butyricum significantly improved neurological deficit, relieved histopathologic change, decreased MDA contents and increased SOD activities in the I/R injury mice. After C. butyricum pretreatment, the expression of Caspase-3 and Bax were significantly decreased, the Bcl-2/Bax ratio was significantly increased, and butyrate contents in the brain were significantly increased. These findings suggested that C. butyricum is able to exert neuroprotective effects against I/R injury mice through anti-oxidant and anti-apoptotic mechanisms, and reversing decrease of butyrate contents in the brain might be involved in its neuroprotection. PMID:26733300

  14. Inhibition of bladder cancer cell proliferation by allyl isothiocyanate (mustard essential oil)

    Sávio, André Luiz Ventura, E-mail: savio.alv@gmail.com [UNESP – Universidade Estadual Paulista, Faculdade de Medicina de Botucatu, Departamento de Patologia, Botucatu, SP (Brazil); Nicioli da Silva, Glenda [UFOP – Universidade Federal de Ouro Preto, Escola de Farmácia, Departamento de Análises Clínicas, Ouro Preto, MG (Brazil); Salvadori, Daisy Maria Fávero [UNESP – Universidade Estadual Paulista, Faculdade de Medicina de Botucatu, Departamento de Patologia, Botucatu, SP (Brazil)

    2015-01-15

    Highlights: • AITC inhibits mutant and wild-type TP53 cell proliferation. • Morphological changes and cells debris were observed after AITC treatment in both cells. • BAX and BCL2 expression modulation was observed in wild-type TP53 cells. • BCL2, BAX and ANLN increased and S100P decreased expression was detected in mutated TP53 cells. • AITC effects in gene modulation are dependent TP53 gene status. - Abstract: Natural compounds hold great promise for combating antibiotic resistance, the failure to control some diseases, the emergence of new diseases and the toxicity of some contemporary medical products. Allyl isothiocyanate (AITC), which is abundant in cruciferous vegetables and mustard seeds and is commonly referred to as mustard essential oil, exhibits promising antineoplastic activity against bladder cancer, although its mechanism of action is not fully understood. Therefore, the aim of this study was to investigate the effects of AITC activity on bladder cancer cell lines carrying a wild type (wt; RT4) or mutated (T24) TP53 gene. Morphological changes, cell cycle kinetics and CDK1, SMAD4, BAX, BCL2, ANLN and S100P gene expression were evaluated. In both cell lines, treatment with AITC inhibited cell proliferation (at 62.5, 72.5, 82.5 and 92.5 μM AITC) and induced morphological changes, including scattered and elongated cells and cellular debris. Gene expression profiles revealed increased S100P and BAX and decreased BCL2 expression in RT4 cells following AITC treatment. T24 cells displayed increased BCL2, BAX and ANLN and decreased S100P expression. No changes in SMAD4 and CDK1 expression were observed in either cell line. In conclusion, AITC inhibits cell proliferation independent of TP53 status. However, the mechanism of action of AITC differed in the two cell lines; in RT4 cells, it mainly acted via the classical BAX/BCL2 pathway, while in T24 cells, AITC modulated the activities of ANLN (related to cytokinesis) and S100P. These data confirm

  15. Apoptosis and the BCL-2 gene family - patterns of expression and prognostic value in STAGE I and II follicular center lymphoma

    Purpose: The prognostic significance of spontaneous levels of apoptosis and Bcl-2, Bax, and Bcl-x protein expression in follicular center lymphoma (FCL) is unknown. The objectives of this retrospective study were (1) to investigate the relationship between pretreatment apoptosis levels and long-term treatment outcome in patients with Stage I and II FCL; (2) to define the incidence and patterns of Bax and Bcl-x protein expression in human FC; and (3) to determine the relationship of Bcl-2, Bax, and Bcl-x expression with spontaneous apoptosis levels and clinical outcome in localized FCL. Methods and Materials: Between 1974 and 1988, 144 patients with Stage I or II FCL were treated. Hematoxylin and eosin (H and E) stained tissue sections of pretreatment specimens were retrieved for 96 patients. Treatment consisted of regional radiation therapy (XRT) for 25 patients, combined modality therapy (CMT) consisting of combination chemotherapy and XRT for 57 patients, and other treatments for 14 patients. Median follow-up for living patients was nearly 12 years. The apoptotic index (AI) was calculated by dividing the number of apoptotic cells by the total number of cells counted and multiplying by 100. Expression of Bcl-2, Bax, and Bcl-x proteins was assessed using immunohistochemistry. Results: The mean and median AI values for the entire group were 0.53 and 0.4, respectively (range: 0-5.2). The AI strongly correlated with cytologic grade, with mean AI values of 0.25 for grade 1, 0.56 for grade 2, and 0.84 for grade 3 (p < 0.0005; Kendall correlation). A positive correlation was present between grouped AI and grouped mitotic index (MI) (p = 0.014). For patients treated with CMT, an AI < 0.4 correlated with improved freedom from relapse (FFR) (p = 0.0145) and overall survival (OS) (p = 0.0081). An AI < 0.4 did not correlate with clinical outcome for the entire cohort or for patients receiving XRT only. Staining of tumor follicles for the Bcl-2 protein was positive, variable

  16. Expression of apoptosis-regulating genes in the rat prostate following botulinum toxin type a injection

    Gorgal Tiago

    2012-01-01

    Full Text Available Abstract Background Onabotulinumtoxin A (OnabotA injection has been investigated as a novel treatment for benign prostatic enlargement caused by benign prostatic hyperplasia. An OnabotA - induced volume reduction caused by sympathetic fibers impairment has been proposed as a potential mechanism of action. Our aim was to investigate the expression of apoptosis-regulating proteins in the rat prostate following OnabotA intraprostatic injection. Methods Adult Wistar rats were injected in the ventral lobes of the prostate with 10 U of OnabotA or saline. A set of OnabotA-injected animals was further treated with 0.5 mg/kg of phenylephrine (PHE subcutaneously daily. All animals were sacrificed after 1 week and had their prostates harvested. Immunohistochemical staining was performed for Bax, Bcl-xL and caspase-3 proteins and visualized by the avidin-biotin method. The optical density of the glandular cells was also determined, with measurement of differences between average optical densities for each group. Results Saline-treated animals showed intense epithelial staining for Bcl-xL and a faint labelling for both Bax and Caspase-3. OnabotA-treated rats showed a reduced epithelial staining of Bcl-xL and a consistently increased Bax and Caspase-3 staining when compared with saline-treated animals. PHE-treated animals showed a stronger Bcl-xL staining and reduced staining of both Bax and Caspase-3 when compared to the OnabotA group. Mean signal intensity measurements for each immunoreaction confirmed a significant decrease of the signal intensity for Bcl-xL and a significant increase of the signal intensity for Bax and Caspase 3 in OnabotA-injected animals when compared with the control group. In OnabotA+PHE treated animals mean signal intensity for Bcl-xL, Bax and Caspase 3 immunoreactions was identical to that of the control animals. Conclusions These results support the hypothesis that OnabotA activates apoptotic pathways in the rat prostate through a

  17. Inhibition of bladder cancer cell proliferation by allyl isothiocyanate (mustard essential oil)

    Highlights: • AITC inhibits mutant and wild-type TP53 cell proliferation. • Morphological changes and cells debris were observed after AITC treatment in both cells. • BAX and BCL2 expression modulation was observed in wild-type TP53 cells. • BCL2, BAX and ANLN increased and S100P decreased expression was detected in mutated TP53 cells. • AITC effects in gene modulation are dependent TP53 gene status. - Abstract: Natural compounds hold great promise for combating antibiotic resistance, the failure to control some diseases, the emergence of new diseases and the toxicity of some contemporary medical products. Allyl isothiocyanate (AITC), which is abundant in cruciferous vegetables and mustard seeds and is commonly referred to as mustard essential oil, exhibits promising antineoplastic activity against bladder cancer, although its mechanism of action is not fully understood. Therefore, the aim of this study was to investigate the effects of AITC activity on bladder cancer cell lines carrying a wild type (wt; RT4) or mutated (T24) TP53 gene. Morphological changes, cell cycle kinetics and CDK1, SMAD4, BAX, BCL2, ANLN and S100P gene expression were evaluated. In both cell lines, treatment with AITC inhibited cell proliferation (at 62.5, 72.5, 82.5 and 92.5 μM AITC) and induced morphological changes, including scattered and elongated cells and cellular debris. Gene expression profiles revealed increased S100P and BAX and decreased BCL2 expression in RT4 cells following AITC treatment. T24 cells displayed increased BCL2, BAX and ANLN and decreased S100P expression. No changes in SMAD4 and CDK1 expression were observed in either cell line. In conclusion, AITC inhibits cell proliferation independent of TP53 status. However, the mechanism of action of AITC differed in the two cell lines; in RT4 cells, it mainly acted via the classical BAX/BCL2 pathway, while in T24 cells, AITC modulated the activities of ANLN (related to cytokinesis) and S100P. These data confirm

  18. Anti-apoptotic effects of aspirin following cerebral ischemia-reperfusion injury in rats

    Liying Qiu; Bin Du; Ying Li; Hongbin Fan; Zhiyong Yang

    2008-01-01

    BACKGROUND: The pharmacological effects of aspirin on apoptosis are complex. The underlying mechanisms have not been properly defined. OBJECTIVE: To observe the effect of different doses of aspirin on brain cell apoptosis following focal cerebral ischemia-reperfusion injury (CIRI) in rats. DESING, TIME AND SETTING: A randomized, controlled, animal experiment, performed at the School of Medicine and Pharmaceutics, Jiangnan University between June and October 2006. MATERIALS: Twenty-six male, adult, Sprague Dawley rats (grade II), weighing 240-290 g, were obtained from Shanghai Experimental Animal Center, Chinese Academy of Sciences. Aspirin was provided by Sigma (USA). METHODS: The rats were randomly divided into four groups: sham-operation (SO), CIRI+ vehicle, CIRI+ aspirin (6 mg/kg), and CIRI + aspirin (60 mg/kg). Rats in the lesion groups were intragastrically administrated saline, aspirin (6 mg/kg), or aspirin (60 mg/kg), respectively. MAIN OUTCOME MEASURES: The number of pyramidal neurons with normal appearance in the cerebral cortex at 2-4 mm from the midline; apoptotic cell death as measured by TUNEL; Bcl-2 and Bax protein localization was determined by immunohistochemistry; maiondiaidehyde (MDA) and super oxidation (SOD) content were determined by biochemistry method; adenosine triphosphate (ATP) content measured by capillary electrophoresis. RESULTS: Following CIRI, the following parameters were altered compared with sham-operated animals: the number of neurons with normal appearance was significantly reduced in the cerebral cortex; the number of apoptotic cells increased; Bax protein expression was enhanced; and the ratio between Bcl-2 and Bax decreased. In addition, MDA content increased significantly, whereas ATP content decreased (P < 0.01 ). Aspirin ameliorated the loss of healthy pyramidal neurons. Both 6 and 60 mg/kg aspirin increased the ratio between Bcl-2 and Bax, with no significant difference between the treatment groups. In addition, 60 mg

  19. The prognostic significance of apoptosis-related biological markers in Chinese gastric cancer patients.

    Xiaowen Liu

    Full Text Available BACKGROUND AND OBJECTIVE: The prognosis varied among the patients with the same stage, therefore there was a need for new prognostic and predictive factors. The aim of this study was to evaluate the relationship of apoptosis-related biological markers such as p53, bcl-2, bax, and c-myc, and clinicopathological features and their prognostic value. METHODS: From 1996 to 2007, 4426 patients had undergone curative D2 gastrectomy for gastric cancer at Fudan University Shanghai Cancer Center. Among 501 patients, the expression levels of p53, bcl-2, bax, and c-myc were examined by immunohistochemistry. The prognostic value of biological markers and the correlation between biological markers and other clinicopathological factors were investigated. RESULTS: There were 339 males and 162 females with a mean age of 57. The percentages of positive expression of p53, bcl-2, bax, and c-myc were 65%, 22%, 43%, and 58%, respectively. There was a strong correlation between p53, bax, and c-myc expression (P=0.00. There was significant association between bcl-2, and bax expression (P<0.05. p53 expression correlated with histological grade (P=0.01; bcl-2 expression with pathological stage (P=0.00; bax expression with male (P=0.02, histological grade (P=0.01, Borrmann type (P=0.01, tumor location (P=0.00, lymph node metastasis (P=0.03, and pathological stage (P=0.03; c-myc expression with Borrmann type (P=0.00. bcl-2 expression was related with good survival in univariate analysis (P=0.01. Multivariate analysis showed that bcl-2 expression and pathological stage were defined as independent prognostic factors. There were significant differences of overall 5-year survival rates according to bcl-2 expression or not in stage IIB (P=0.03. CONCLUSION: The expression of bcl-2 was an independent prognostic factor for patients with gastric cancer; it might be a candidate for the gastric cancer staging system.

  20. Protective Effects of N-Acetyl-L-cystein on 3,4-Methylene Dioxymethamphetamie-Induced Neurotoxicity in Cerebellum of Male Rats

    Sara Soleimani Asl

    2011-10-01

    Full Text Available Objective(s: 3-4, methylenedioxymethamphetamine (MDMA causes apoptosis in nervous system and several studies suggest that oxidative stress contributes to MDMA-induced neurotoxicity. The aim of this study is to examine the effects of N-acetyl-L-Cystein (NAC as an antioxidant on MDMA-induced apoptosis. Materials and Methods: 21 Sprague dawley male rats (200-250mg were treated with MDMA (2×0,5mg/kg or MDMA plus NAC (100mg/kg IP for 7 day. After last administration of MDMA, rats were killed, cerebellum was removed and Bax and Bcl-2 expression was assessed by western blotting method. Results: The results of this study showed that MDMA causes up-regulation of Bax and down-regulation of Bcl-2 and NAC administration attenuated MDMA-induced apoptosis. Conclusion: The present study suggests that NAC treatment may improve MDMA-induced neurotoxicity.

  1. Apolipoprotein E mimetic peptide protects against diffuse brain injur y

    Yaning Zhao; Jianmin Li; Qiqun Tang; Junling Gao; Changxiang Chen; Liwei Jing; Pan Zhang; Shuxing Li

    2014-01-01

    Apolipoprotein E plays a crucial role in inhibiting chronic neurodegenerative processes. Howev-er, its impact on neurological function following diffuse brain injury is still unclear. This study was designed to evaluate the therapeutic effects and mechanisms of action of apolipoprotein E mimetic peptide on diffuse brain injury. Apolipoprotein E mimetic peptide was administered into the caudal vein of rats with diffuse brain injury before and after injury. We found that apo-lipoprotein E mimetic peptide signiifcantly decreased the number of apoptotic neurons, reduced extracellular signal-regulated kinase1/2 phosphorylation, down-regulated Bax and cytochrome c expression, decreased malondialdehyde content, and increased superoxide dismutase activity in a dose-dependent manner. These experimental ifndings demonstrate that apolipoprotein E mimetic peptide improves learning and memory function and protects against diffuse brain injury-induced apoptosis by inhibiting the extracellular signal-regulated kinase1/2-Bax mito-chondrial apoptotic pathway.

  2. Heptachlor induced mitochondria-mediated cell death via impairing electron transport chain complex III

    Highlights: •Heptachlor inhibited mitochondrial electron transport chain complex III activity. •Heptachlor promoted generation of reactive oxygen species. •Heptachlor induced Bax activation. •Heptachlor induced mitochondria-mediated and caspase-dependent apoptosis. -- Abstract: Environmental toxins like pesticides have been implicated in the pathogenesis of Parkinson’s disease (PD). Epidemiological studies suggested that exposures to organochlorine pesticides have an association with an increased PD risk. In the present study, we examined the mechanism of toxicity induced by an organochlorine pesticide heptachlor. In a human dopaminergic neuroblastoma SH-SY5Y cells, heptachlor induced both morphological and functional damages in mitochondria. Interestingly, the compound inhibited mitochondrial electron transport chain complex III activity. Rapid generation of reactive oxygen species and the activation of Bax were then detected. Subsequently, mitochondria-mediated, caspase-dependent apoptosis followed. Our results raise a possibility that an organochlorine pesticide heptachlor can act as a neurotoxicant associated with PD

  3. STAT5 activation by human GH protects insulin-producing cells against interleukin-1beta, interferon-gamma and tumour necrosis factor-alpha-induced apoptosis independent of nitric oxide production

    Jensen, Janne; Galsgaard, Elisabeth D; Karlsen, Allan E;

    2005-01-01

    proliferation and insulin production in pancreatic beta-cells and rat insulin-producing INS-1 cells. Here we report that human (h) GH can prevent the apoptotic effects of IL-1beta, IFN-gamma and TNF-alpha in INS-1 and INS-1E cells. Using adenovirus-mediated gene transfer, we found that the anti-apoptotic effect...... expression and in fact augmented the IL-1beta-stimulated nitric oxide production. As the anti-apoptotic Bcl-xL gene has been shown to harbour a STAT5-binding element we measured the expression of Bcl-xL as well as the pro-apoptotic Bax. We found that hGH increased the Bcl-xL/Bax ratio both in the absence and...

  4. Curcumin inhibits the proliferation of a human colorectal cancer cell line Caco-2 partially by both apoptosis and G2/M cell cycle arrest

    Yohko Fujimoto

    2014-06-01

    Full Text Available The aim of this study was to assess the possible roles of the phytochemical compounds, curcumin, quercetin and resveratrol in the proliferation of human colorectal cancer cell line Caco-2. All three phytochemical compounds inhibited Caco-2 cell proliferation, with curcumin being more effective than quercetin and resveratrol. Investigations concerning DNA fragmentation in the nucleus, Bax and Bcl-2 mRNA expression levels, and caspase-3/7 activity indicated that curcumin induced apoptosis in Caco-2 cells through an increase in the Bax/Bcl-2 ratio and activation of caspase-3/7. Furthermore, the analysis of flow-cytometry showed that curcumin caused an arrest of G2/M phase in Caco-2 cells. These results suggest that curcumin suppresses Caco-2 proliferation partially via activation of the mitochondrial apoptotic pathway and cell cycle retardation.

  5. Acupuncture at Baihui and Dazhui reduces brain cell apoptosis in heroin readdicts

    Xiaorong Hou; Rongjun Zhang; Hang Lv; Xinghui Cai; Guangchuan Xie; Xiaoge Song

    2014-01-01

    Acupuncture at Baihui (GV20) and Dazhui (GV14) reduces neuronal loss and attenuates ultra-structural damage in cerebral ischemic rats. However, whether acupuncture can treat addiction and prevent readdiction through changes to brain cell ultrastructure remains unknown. In this study, cell apoptosis was observed in the hippocampus and frontal lobe of heroin readdicted rats by electron microscopy. Immunohistochemical staining displayed a reduction in Bcl-2 ex-pression and an increase in Bax expression in the hippocampus and frontal lobe. After rats were given acupuncture at Baihui and Dazhui, the pathological damage in the hippocampus and frontal lobe was signiifcantly reduced, Bcl-2 expression was upregulated and Bax expression was downregulated. Acupuncture exerted a similar effect with methadone, a commonly used drug for clinical treatment of drug addiction. Experimental ifndings suggest that acupuncture at Dazhui and Baihui can prevent brain cell apoptosis in heroin readdicted rats.

  6. Measurement of subcellular texture by optical Gabor-like filtering with a digital micromirror device

    Pasternack, Robert M.; Qian, Zhen; Zheng, Jing-Yi; Metaxas, Dimitris N.; White, Eileen; Boustany, Nada N.

    2010-01-01

    We demonstrate an optical Fourier processing method to quantify object texture arising from subcellular feature orientation within unstained living cells. Using a digital micromirror device as a Fourier spatial filter, we measured cellular responses to two-dimensional optical Gabor-like filters optimized to sense orientation of nonspherical particles, such as mitochondria, with a width around 0.45 μm. Our method showed significantly rounder structures within apoptosis-defective cells lacking the proapoptotic mitochondrial effectors Bax and Bak, when compared with Bax/Bak expressing cells functional for apoptosis, consistent with reported differences in mitochondrial shape in these cells. By decoupling spatial frequency resolution from image resolution, this method enables rapid analysis of nonspherical submicrometer scatterers in an under-sampled large field of view and yields spatially localized morphometric parameters that improve the quantitative assessment of biological function. PMID:18830354

  7. Heptachlor induced mitochondria-mediated cell death via impairing electron transport chain complex III

    Hong, Seokheon; Kim, Joo Yeon; Hwang, Joohyun [Department of Molecular Biology, Sejong University, Seoul 143-747 (Korea, Republic of); Shin, Ki Soon [Department of Biology, Department of Life and Nanopharmaceutical Sciences, Kyung Hee University, Seoul 130-701 (Korea, Republic of); Kang, Shin Jung, E-mail: sjkang@sejong.ac.kr [Department of Molecular Biology, Sejong University, Seoul 143-747 (Korea, Republic of)

    2013-08-09

    Highlights: •Heptachlor inhibited mitochondrial electron transport chain complex III activity. •Heptachlor promoted generation of reactive oxygen species. •Heptachlor induced Bax activation. •Heptachlor induced mitochondria-mediated and caspase-dependent apoptosis. -- Abstract: Environmental toxins like pesticides have been implicated in the pathogenesis of Parkinson’s disease (PD). Epidemiological studies suggested that exposures to organochlorine pesticides have an association with an increased PD risk. In the present study, we examined the mechanism of toxicity induced by an organochlorine pesticide heptachlor. In a human dopaminergic neuroblastoma SH-SY5Y cells, heptachlor induced both morphological and functional damages in mitochondria. Interestingly, the compound inhibited mitochondrial electron transport chain complex III activity. Rapid generation of reactive oxygen species and the activation of Bax were then detected. Subsequently, mitochondria-mediated, caspase-dependent apoptosis followed. Our results raise a possibility that an organochlorine pesticide heptachlor can act as a neurotoxicant associated with PD.

  8. Protective effects of Purendan superfine powder on retinal neuron apoptosis in a rat model of type 2 diabetes mellitus

    Zhijun Dong; Xiangyi Tao; Xiaoxiao Fu; Haibin Wang; Donghua Wang; Tiemin Zhang

    2012-01-01

    This study sought to investigate the effects of Purendan superfine powder comprised of Momordica charantia, Radix Ginseng, and Radix Salviae Miltiorrhiae on neuronal apoptosis and expression of bcl-2, bax, and caspase-3, which are retinal apoptosis-associated factors in rats with diabetes mellitus induced by continuous intraperitoneal injection of streptozotocin. The results showed that Purendan superfine powder could upregulate the expression of bcl-2 protein and mRNA, and downregulate the expression of bax and caspase-3 in the retina of diabetes mellitus rats. In addition, Purendan superfine powder was shown to reduce the number of apoptotic neurons. Our experimental findings indicate that Purendan superfine powder can inhibit neuronal apoptosis in the retina of diabetes mellitus rats and has protective effects on diabetic retinopathy.

  9. Data on four apoptosis-related genes in the colonial tunicate Botryllus schlosseri.

    Franchi, Nicola; Ballin, Francesca; Manni, Lucia; Schiavon, Filippo; Ballarin, Loriano

    2016-09-01

    The data described are related to the article entitled "Recurrent phagocytosis-induced apoptosis in the cyclical generation change of the compound ascidian Botryllus schlosseri" (Franchi et al., 2016) [1]. Four apoptosis-related genes, showing high similarity with mammalian Bax (a member of the Bcl-2 protein family), AIF1 (apoptosis-inducing factor-1), PARP1 (poly ADP ribose polymerase-1) and IAP7 (inhibitor of apoptosis-7) were identified from the analysis of the trascriptome of B. schlosseri. They were named BsBax, BsAIF1, BsPARP1 and BsIAP7. Here, their deduced amino acid sequence were compared with known sequences of orthologous genes from other deuterostome species together with a study of their identity/similarity. PMID:27294183

  10. The morphological changes in transplanted tumors in rats at plasmonic photothermal therapy

    Bucharskaya, Alla B.; Maslyakova, Galina N.; Navolokin, Nikita A.; Dikht, Nataliya I.; Terentyuk, Georgy S.; Bashkatov, Alexey N.; Genina, Elina A.; Khlebtsov, Boris N.; Khlebtsov, Nikolai G.; Tuchin, Valery V.

    2016-04-01

    The aim of work was to study the morphological changes in transplanted liver tumors of rats after plasmonic photothermal therapy (PPTT). The gold nanorods functionalized with thiolated polyethylene glycol were injected intravenously to rats with transplanted liver cancer PC-1. A day after injection the tumors were irradiated by the infrared 808-nm diode laser. The withdrawal of the animals from the experiment and sampling of tumor tissue for morphological study were performed 24 hours after the laser exposure. The standard histological and immunohistochemical staining with antibodies to proliferation marker Ki-67 and apoptosis marker BAX were used for morphological study of transplanted tumors. The plasmonic photothermal therapy had pronounced damaging effect in rats with transplanted liver tumors expressed in degenerative and necrotic changes in the tumor cells. The decrease of proliferation marker Ki-67 and increase of expression of apoptosis marker BAX were observed in tumor cells after PPTT.

  11. Lipolysis and apoptosis of adipocytes induced by neuropeptide Y—Y5 receptor antisense oligodeoxynucleotides in obese rats

    GONGHai-Xia; GUOXi-Rong; FEILi; GUOMei; LIUQian-Qi; CHENRong-Hua

    2003-01-01

    AIM:To investigate the influence of central administration of neuropeptide Y-Y5 receptor antisense oligodeoxynucleotides(ODN) on the body weight and fat pads of high-energy diet-induced obese rats, and the effects on white adipocyte lipolysis and apoptosis. METHODS: Y5 receptor antisense, sense, mismatched oligodeoxynucleotides (ODN) or vehicle were intracerebroventricularly injected, and average adipocyte area was calculated. DNA ladders were measured to evaluate adipocyte apoptosis, and RT-PCR was used to analyze the expression of bcl-2 and bax gene. RESULTS: (1) Central administration of Y5 receptor antisense ODN significantly decreased body weight, fat pads, and average adipocyte area. (2) DNA fragmentation was presented after electrophoresis at both epididymal and retroperitoneal adipose tissue. (3) The expression of bcl-2 gene was downregulated, while the expression of bax was upregulated. CONCLUSION:Lipolysis and adipocyte apoptosis may be important reasons for Y5 receptor antisense therapy.

  12. A study on apoptotic signaling pathway in HL-60 cells induced by radiation

    The mechanical insights of death at cancer cells by ionizing radiation are not yet clearly defined. Recent evidences have demonstrated that radiation therapy may induce cell death via activation of signaling pathway for apoptosis in target cells. This study is designed whether ionizing radiation may activate the signaling cascades of apoptosis including caspase family cysteine proteases, Bcl2/Bax, cytochrome c and Fas/Fas-L in target cells. HL-60 cells were irradiated in vitro with 6 MV X-ray at dose ranges from 2 Gy to 32 Gy. The cell viability was tested by MTT assay and the extent of apoptosis was determined using agarose gel electrophoresis. The activities of caspase proteases were measured by proteolytic cleavages of substrates. Western blot analysis was used to monitor PARP, caspase-3, Cytochrome-c, BcI-2, Bax, Fas and Fas-L. Ionizing radiation decreases the viability of HL -60 cells in a time and dose dependent manner. Ionizing radiation-induced death in HL- 60 cells is an apoptotic death which is revealed as characteristic ladder-pattern fragmentation at genomic DNA over 16 Gy at 4 hours. Ionizing radiation induces the activation of caspase-2, 3, 6, 8 and 9 of HL --60 cells in a time-dependent manner. The activation of caspase- 3 protease is also evidenced by the digestion of poly (ADP-ribose) polymerase and procaspase 3 with 16Gy ionizing irradiation. Anti-apoptotic Bcl2 expression is decreased but apoptotic Bax expression is increased with mitochondrial cytochrome c release in a time- dependent manner. In addition, expression of Fas and Fas-L is also increased in a time dependent manner. These data suggest that ionizing radiation-induced apoptosis is mediated by the activation of various signaling pathways including caspase family cysteine proteases, BcI2/Bax, Fas and Fas-L in a time and dose dependent manner

  13. Neuroprotection of aucubin in primary diabetic encephalopathy

    2008-01-01

    Hippocampal neuronal apoptosis accompanied by impairment of cognitive function occurs in primary diabetic encephalopathy. In this study, we investigated the neuroprotective mechanism of the iridoid glycoside, aucubin, using rats (n=8). Diabetes mellitus was induced in the rats by intraperitoneal (i.p.) injection of streptozotocin (60 mg/kg body weight). After 65 d, half of the DM rats were administered aucubin (5 mg/kg; i.p.) for 15 d, yielding treatment DM+A. A third group of rats received no strepto- zotocin or aucibin, and served as controls (CON). Encephalopathy was assessed using Y-maze be- havioral testing. Rats were euthanized on Day 87, and hippocampi were excised for visual (light and transmission electron microscopic) and immunochemical (Western blot; immunohistochemical) as- sessments of the CA1 subfield for apoptosis and expression of regulatory proteins Bcl-2 and Bax. Treatment responses to all the parameters examined (body weight, plasma glucose, Y-maze error rates, pyramidal cell ultrastructure, proportions of apoptotic cells, levels of expression of Bcl-2 and Bax, and survivability of neuronal cells) were identical: there were highly significant differences between DM and CON groups (P<0.001), but the effects were significantly moderated (P<0.01) in DM+A compared with DM. These findings confirm the association of apoptosis with the encephalopathic effects of diabetes mellitus, and suggest a major role of the expression levels of Bcl-2 and Bax in the regulation of apop- totic cell death. All of the results suggest that aucubin could effectively inhibit apoptosis by modulating the expressions of Bcl-2 and Bax genes.

  14. Neuroprotection of aucubin in primary diabetic encephalopathy

    XUE HongYu; JIN LiJi; JIN Lei; ZHANG Peng; LI DanQing; XIA YanQiu; LU YaNan; XU YongPing

    2008-01-01

    Hippocampal neuronal apoptosis accompanied by impairment of cognitive function occurs in primary diabetic encephalopathy. In this study, we investigated the neuroprotective mechanism of the iridoid glycoside, aucubin, using rats (n=8). Diabetes mellitus was induced in the rats by intraperitoneal (i.p.) injection of streptozotocin (60 mg/kg body weight). After 65 d, half of the DM rats were administered aucubin (5 mg/kg;i.p.) for 15 d, yielding treatment DM+A. A third group of rats received no strepto-zotocin or aucibin, and served as controls (CON). Encephalopathy was assessed using Y-maze be-havioral testing. Rats were euthanized on Day 87, and hippocampi were excised for visual (light and transmission electron microscopic) and immunochemical (Western blot;immunohistochemical) as-sessments of the CA1 subfield for apoptosis and expression of regulatory proteins Bcl-2 and Bax. Treatment responses to all the parameters examined (body weight, plasma glucose, Y-maze error rates, pyramidal cell ultrastructure, proportions of apoptotic cells, levels of expression of Bcl-2 and Bax, and survivability of neuronal cells) were identical: there were highly significant differences between DM and CON groups (P<0.001), but the effects were significantly moderated (P<0.01) in DM+A compared with DM. These findings confirm the association of apoptosis with the encephalopathic effects of diabetes mellitus, and suggest a major role of the expression levels of Bcl-2 and Bax in the regulation of apop-totic cell death. All of the results suggesf that aucubin could effectively inhibit apoptosis by modulating the expressions of Bcl-2 and Bax genes.

  15. A Test of Abelson and Baysinger's (1984) Optimal Turnover Hypothesis in the Context of Public Organizations using Computational Simulation

    Kohn, Harold D.

    2008-01-01

    Both practitioners and researchers have long noted that employee turnover creates both positive and negative consequences for an organization. From a management perspective, the question is how much turnover is the right amount. Abelson and Baysinger (1984) first proposed that an optimal level of turnover could be found based on individual, organizational, and environmental factors. However, as Glebbeek and Bax (2004) noted, their approach was overly complex to empirically verify, let alon...

  16. Decreased Apoptotic Rate of Alveolar Macrophages of Patients with Idiopathic Pulmonary Fibrosis

    Fotios Drakopanagiotakis; Areti Xifteri; Evaggelos Tsiambas; Andreas Karameris; Konstantina Tsakanika; Napoleon Karagiannidis; Demetrios Mermigkis; Vlasis Polychronopoulos; Demosthenes Bouros

    2012-01-01

    Introduction. Increased apoptosis of epithelial cells and decreased apoptosis of myofibroblasts are involved in the pathogenesis of IPF. The apoptotic profile of alveolar macrophages (AMs) in IPF is unclear. Aim. To investigate whether AMs of patients with IPF exhibit a different apoptotic profile compared to normal subjects. Methods. We analyzed, by immunohistochemistry, the expression of the apoptotic markers fas, fas ligand , bcl-2, and bax in AM obtained from bronchoalveolar lavage fluid ...

  17. Thiol protecting agents and antioxidants inhibit the mitochondrial permeability transition promoted by etoposide: implications in the prevention of etoposide-induced apoptosis

    Custódio, José B. A.; Cardoso, Carla M. P.; Almeida, Leonor M.

    2002-01-01

    Etoposide (VP-16) is known to promote cell apoptosis either in cancer or in normal cells as a side effect. This fact is preceded by the induction of several mitochondrial events, including increase in Bax/Bcl-2 ratio followed by cytochrome c release and consequent activation of caspase-9 and -3, reduction of ATP levels, depolarization of membrane potential ([Delta][Psi]) and rupture of the outer membrane. These events are apoptotic factors essentially associated with the induction of the mito...

  18. Beta-blockers and health-related quality of life in patients with peripheral arterial disease and COPD

    Yvette RBM van Gestel; Sanne E Hoeks; Don D Sin; Henk Stam; Frans W Mertens; Bax, Jeroen J.; van Domburg, Ron T.; Don Poldermans

    2009-01-01

    Yvette RBM van Gestel1, Sanne E Hoeks1, Don D Sin2, Henk Stam3, Frans W Mertens3, Jeroen J Bax4, Ron T van Domburg5, Don Poldermans61Department of Anesthesiology, Erasmus Medical Center, Rotterdam, The Netherlands; 2Department of Medicine, University of British Columbia and The James Hogg iCAPTURe Center, St. Paul’s Hospital, Vancouver, Canada; 3Department of Pulmonology, Erasmus Medical Center, Rotterdam, The Netherlands; 4Department of Cardiology, Leiden University Medica...

  19. Evaluation of In Vitro Anticancer Activity of Ocimum Basilicum, Alhagi Maurorum, Calendula Officinalis and Their Parasite Cuscuta Campestris

    Mandana Behbahani

    2014-01-01

    The present investigation was carried out to study the relationship between presence of cytotoxic compounds in Ocimum basilicum, Alhagi maurorum, Calendula officinalis and their parasite Cuscuta campestris. The cytotoxic activity of the pure compounds was performed by MTT assay against breast cancer cell lines (MCF-7 and MDA-MB-231) and normal breast cell line (MCF 10A). The induction of apoptosis was measured by the expression levels of p53, bcl-2, bax and caspase-3 genes using quantitative ...

  20. Arsenic Trioxide Induces Apoptosis in Human Platelets via C-Jun NH2-Terminal Kinase Activation

    Yicun Wu; Jin Dai; Weilin Zhang; Rong Yan; Yiwen Zhang; Changgeng Ruan; Kesheng Dai

    2014-01-01

    Arsenic trioxide (ATO), one of the oldest drugs in both Western and traditional Chinese medicine, has become an effective anticancer drug, especially in the treatment of acute promyelocytic leukemia (APL). However, thrombocytopenia occurred in most of ATO-treated patients with APL or other malignant diseases, and the pathogenesis remains unclear. Here we show that ATO dose-dependently induces depolarization of mitochondrial inner transmembrane potential (ΔΨm), up-regulation of Bax and down-re...

  1. Evolution of Eutrophication depending on Environmental Conditions: A Case Study in a Reservoir.

    Xana Álvarez; Enrique Valero; Juan Picos

    2014-01-01

    In the last years, the phenomenon of eutrophication is affecting many river ecosystems. Besides being an environmental problem, it can also be a problem for human health. This study aims to explore how environmental parameters affect the concentration of green algae and cyanobacteria throughout the period from 2010 to 2013 in A Baxe reservoir (NW Spain). Factors such as temperature, rainfall and solar radiation influenced in the presence of the algae, as well as in its concentration. Being li...

  2. Antisense bcl-2 treatment increases programmed cell death in non-small cell lung cancer cell lines.

    Koty, P P; Zhang, H; Levitt, M L

    1999-02-01

    Programmed cell death (PCD) is a genetically regulated pathway that is altered in many cancers. This process is, in part, regulated by the ratio of PCD inducers (Bax) or inhibitors (Bcl-2). An abnormally high ratio of Bcl-2 to Bax prevents PCD, thus contributing to resistance to chemotherapeutic agents, many of which are capable of inducing PCD. Non-small cell lung cancer (NSCLC) cells demonstrate resistance to these PCD-inducing agents. If Bcl-2 prevents NSCLC cells from entering the PCD pathway, then reducing the amount of endogenous Bcl-2 product may allow these cells to spontaneously enter the PCD pathway. Our purpose was to determine the effects of bcl-2 antisense treatment on the levels of programmed cell death in NSCLC cells. First, we determined whether bcl-2 and bax mRNA were expressed in three morphologically distinct NSCLC cell lines: NCI-H226 (squamous), NCI-H358 (adenocarcinoma), and NCI-H596 (adenosquamous). Cells were then exposed to synthetic antisense bcl-2 oligonucleotide treatment, after which programmed cell death was determined, as evidenced by DNA fragmentation. Bcl-2 protein expression was detected immunohistochemically. All three NSCLC cell lines expressed both bcl-2 and bax mRNA and had functional PCD pathways. Synthetic antisense bcl-2 oligonucleotide treatment resulted in decreased Bcl-2 levels, reduced cell proliferation, decreased cell viability, and increased levels of spontaneous PCD. This represents the first evidence that decreasing Bcl-2 in three morphologically distinct NSCLC cell lines allows the cells to spontaneously enter a PCD pathway. It also indicates the potential therapeutic use of antisense bcl-2 in the treatment of NSCLC. PMID:10217615

  3. Mutual regulation of Bcl-2 proteins independent of the BH3 domain as shown by the BH3-lacking protein Bcl-x(AK.

    Michael Plötz

    Full Text Available The BH3 domain of Bcl-2 proteins was regarded as indispensable for apoptosis induction and for mutual regulation of family members. We recently described Bcl-x(AK, a proapoptotic splice product of the bcl-x gene, which lacks BH3 but encloses BH2, BH4 and a transmembrane domain. It remained however unclear, how Bcl-x(AK may trigger apoptosis.For efficient overexpression, Bcl-x(AK was subcloned in an adenoviral vector under Tet-OFF control. The construct resulted in significant apoptosis induction in melanoma and nonmelanoma cell lines with up to 50% apoptotic cells as well as decreased cell proliferation and survival. Disruption of mitochondrial membrane potential, and cytochrome c release clearly indicated activation of the mitochondrial apoptosis pathways. Both Bax and Bak were activated as shown by clustering and conformation analysis. Mitochondrial translocation of Bcl-x(AK appeared as an essential and initial step. Bcl-x(AK was critically dependent on either Bax or Bak, and apoptosis was abrogated in Bax/Bak double knockout conditions as well by overexpression of Bcl-2 or Bcl-x(L. A direct interaction with Bcl-2, Bax, Bad, Noxa or Puma was however not seen by immunoprecipitation. Thus besides BH3-mediated interactions, there exists an additional way for mutual regulation of Bcl-2 proteins, which is independent of the BH3. This pathway appears to play a supplementary role also for other proapoptotic family members, and its unraveling may help to overcome therapy resistance in cancer.

  4. Multimodal Interaction with BCL-2 Family Proteins Underlies the Pro-Apoptotic Activity of PUMA BH3

    Edwards, Amanda L.; Gavathiotis, Evripidis; LaBelle, James L.; Braun, Craig R.; Opoku-Nsiah, Kwadwo A.; Bird, Gregory H.; Walensky, Loren D.

    2013-01-01

    PUMA is a pro-apoptotic BCL-2 family member that drives the apoptotic response to a diversity of p53-dependent and independent cellular insults. Deciphering the spectrum of PUMA interactions that confer its context-dependent pro-apoptotic properties remains a high priority goal. Here, we report the synthesis of PUMA SAHBs, structurally-stabilized PUMA BH3 helices that, in addition to broadly targeting anti-apoptotic proteins, directly bind to BAX. NMR, photocrosslinking, and biochemical analy...

  5. One-step generation of triple knockout CHO cell lines using CRISPR/Cas9 and fluorescent enrichment

    Grav, Lise Marie; Lee, Jae Seong; Thomsen, Signe Gerling;

    2015-01-01

    The CRISPR/Cas9 genome editing technology has previously been shown to be a highly efficient tool for generating gene disruptions in CHO cells. In this study we further demonstrate the applicability and efficiency of CRISPR/Cas9 genome editing by disrupting FUT8, BAK and BAX simultaneously in a....... Taken together, multiplexing with CRISPR/Cas9 can accelerate genome engineering efforts in CHO cells even further....

  6. Targeting Antiapoptotic Bcl-2 Family Members with Cell-Permeable BH3 Peptides Induces Apoptosis Signaling, Death in Head, Neck Squamous Cell Carcinoma Cells

    Rongxiu Li

    2007-10-01

    Full Text Available Head, neck squamous cell carcinomas (HNSCCs are frequently characterized by chemotherapy, radiation resistance, by overexpression of Bcl-XL, an antiapoptotic member of the Bcl-2 protein family. In this report, we examined whether cell-permeable peptides derived from the BH3 domains of proapoptotic Bax, Bad, or Bak could be used to target Bcl-XL and/or Bcl-2 in HNSCC cells, induce apoptotic death in these cells. To render the peptides cell-permeable, Antennapedia (Ant or polyarginine (R8 peptide transduction domain was fused to the amino termini. Fluorescence microscopy of peptide-treated HNSCC cells revealed that the BH3 peptides colocalized with mitochondria, the site of Bcl-XL, Bcl-2 expression. By contrast, a mutant peptide (BaxE BH3 that cannot bind Bcl-XL or Bcl-2 was diffusely localized throughout the cytoplasm. Treatment of three HNSCC cell lines (1483, UM-22A, UM-22B with the wild-type BH3 peptides resulted in loss of viability, induction of apoptosis, as assessed by 3-(4,5-dimethythiazol-2yl-5-(3-carboxymethoxyphenyl-2-(4-sulfophenyl-2H-tetrazolium (MTS assays, annexin V staining. In general, Ant-conjugated peptides were more potent than R8-conjugated peptides, Bad BH3 peptide was typically more potent than Bax BH3 or Bak BH3. Treatment of purified HNSCC mitochondria with BH3 peptides resulted in robust release of cytochrome c. Thus, the relative apoptosis resistance of HNSCC cells is not due to a deficit in this step of the intrinsic, mitochondrialmediated apoptosis pathway. We conclude that cellpermeable BH3 peptides can be used to target Bcl-XL and/or Bcl-2 in HNSCC, that targeting of these proteins may have therapeutic value in the treatment of this disease.

  7. Edaravone protects PC12 cells from ischemic-like injury via attenuating the damage to mitochondria

    SONG Ying; LI Meng; LI Ji-cheng; WEI Er-qing

    2006-01-01

    Background: Edaravone had been validated to effectively protect against ischemic injuries. In this study, we investigated the protective effect of edaravone by observing the effects on anti-apoptosis, regulation of Bcl-2/Bax protein expression and recovering from damage to mitochondria after OGD (oxygen-glucose deprivation)-reperfusion. Methods: Viability of PC 12cells which were injured at different time of OGD injury, was quantified by measuring MTT (2-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) staining. In addition, PC 12 cells' viability was also quantified after their preincubation in different concentration of edaravone for 30 min followed by (OGD). Furthermore, apoptotic population of PC12 cells that reinsulted from OGD-reperfusion with or without preincubation with edaravone was determined by flow cytometer analysis,electron microscope and Hoechst/PI staining. Finally, change of Bcl-2/Bax protein expression was detected by Westem blot.Results: (1) The viability of PC 12 cells decreased with time (1~12 h) after OGD. We regarded the model of OGD 2 h, then replacing DMEM (Dulbecco's Modified Eagle's Medium) for another 24 h as an OGD-reperfusion in this research. Furthermore,most PC12 cells were in the state of apoptosis after OGD-reperfusion. (2) The viability of PC12 cells preincubated with edaravone at high concentrations (1,0.1, 0.01 μmol/L) increased significantly with edaravone protecting PC 12 cells from apoptosis after OGD-reperfusion injury. (3) Furthermore, edaravone attenuates the damage of OGD-reperfusion on mitochondria and regulated Bcl-2/Bax protein imbalance expression after OGD-reperfusion. Conclusion: Neuroprotective effects of edaravone on ischemic or other brain injuries may be partly mediated through inhibition of Bcl-2/Bax apoptotic pathways by recovering from the damage of mitochondria.

  8. Megakaryocytes possess a functional intrinsic apoptosis pathway that must be restrained to survive and produce platelets

    Josefsson, Emma C.; James, Chloé; Henley, Katya J.; Debrincat, Marlyse A.; Rogers, Kelly L.; Dowling, Mark R; Michael J White; Kruse, Elizabeth A; Lane, Rachael M.; Ellis, Sarah; Nurden, Paquita; Mason, Kylie D; O’Reilly, Lorraine A; Roberts, Andrew W.; Metcalf, Donald

    2011-01-01

    It is believed that megakaryocytes undergo a specialized form of apoptosis to shed platelets. Conversely, a range of pathophysiological insults, including chemotherapy, are thought to cause thrombocytopenia by inducing the apoptotic death of megakaryocytes and their progenitors. To resolve this paradox, we generated mice with hematopoietic- or megakaryocyte-specific deletions of the essential mediators of apoptosis, Bak and Bax. We found that platelet production was unperturbed. In stark cont...

  9. Synergistic interactions between HDAC and sirtuin inhibitors in human leukemia cells.

    Michele Cea

    Full Text Available Aberrant histone deacetylase (HDAC activity is frequent in human leukemias. However, while classical, NAD(+-independent HDACs are an established therapeutic target, the relevance of NAD(+-dependent HDACs (sirtuins in leukemia treatment remains unclear. Here, we assessed the antileukemic activity of sirtuin inhibitors and of the NAD(+-lowering drug FK866, alone and in combination with traditional HDAC inhibitors. Primary leukemia cells, leukemia cell lines, healthy leukocytes and hematopoietic progenitors were treated with sirtuin inhibitors (sirtinol, cambinol, EX527 and with FK866, with or without addition of the HDAC inhibitors valproic acid, sodium butyrate, and vorinostat. Cell death was quantified by propidium iodide cell staining and subsequent flow-cytometry. Apoptosis induction was monitored by cell staining with FITC-Annexin-V/propidium iodide or with TMRE followed by flow-cytometric analysis, and by measuring caspase3/7 activity. Intracellular Bax was detected by flow-cytometry and western blotting. Cellular NAD(+ levels were measured by enzymatic cycling assays. Bax was overexpressed by retroviral transduction. Bax and SIRT1 were silenced by RNA-interference. Sirtuin inhibitors and FK866 synergistically enhanced HDAC inhibitor activity in leukemia cells, but not in healthy leukocytes and hematopoietic progenitors. In leukemia cells, HDAC inhibitors were found to induce upregulation of Bax, a pro-apoptotic Bcl2 family-member whose translocation to mitochondria is normally prevented by SIRT1. As a result, leukemia cells become sensitized to sirtuin inhibitor-induced apoptosis. In conclusion, NAD(+-independent HDACs and sirtuins cooperate in leukemia cells to avoid apoptosis. Combining sirtuin with HDAC inhibitors results in synergistic antileukemic activity that could be therapeutically exploited.

  10. Mean-Field Calculations for the Three-Dimensional Holstein Model

    罗强; 刘川

    2002-01-01

    The electron-phonon Holstein model is studied in three spatial dimensions. It is argued that this model can be used to account for major features of the high-To BaPb1-xBixO3 and BaxK1-xBiO3 systems. Mean-field calculations are performed via a path integral representation of the model. Charge-density-wave order parameters and transition temperatures are obtained.

  11. A novel chenodeoxycholic derivative HS-1200 enhances radiation-induced apoptosis in human MCF-7 breast cancer cells

    To examine whether a synthetic bile acid derivatives (HS-1200) sensitizes the radiation-induced apoptosis in human breast cancer cells (MCF-7) and to investigate the underlying mechanism. Human breast cancer cells (MCF-7) in exponential growth phase were treated with HS-1200 for 24 hours at 37 .deg. C with 5% CO2 in air atmosphere. After removal of HS-1200, cells were irradiated with 2 ∼ 8 Gy X-ray, and then cultured in drug-free media for 24-96 hours. The effect of radiation on the clonogenicity of MCF-7 cells was determined with clonogenic cell survival assay with 16 μ M of HS-1200. The induction of apoptosis was determined using agarose gel electrophoresis and Hoechst staining. The expression level of apoptosis-related molecules, such as PARP, Bax, Bcl-2, Bak and AIF, were assayed by Western blotting analysis with 40 μ M of HS-1200 combined with 8 Gy irradiation. To examine the cellular location of cytochrome c, bax and AIF immunofluorescent stainings were undertaken. Treatment of MCF-7 cells with 40 μ M of HS-1200 combined with 8 Gy irradiation showed several changes associated with enhanced apoptosis by agarose gel electrophoresis and Hoechst staining. HS-1200 combined with 8 Gy irradiation treatment also enhanced production of PARP cleavage products and increased Bax/Bcl-2 ratio by Western blotting. Loss of mitochondrial membrane potential (Δ ψ m) and increased cytochrome c staining indicated that cytochrome c had been released from the mitochondria in HS-1200 treated cells. We demonstrated that combination treatment with a synthetic chenodeoxycholic acid derivative HS-1200 and irradiation enhanced radiation-induced apoptosis of human breast cancer cells (MCF-7). We suggest that the increased Bax/Bcl-2 ratio in HS-1200 co-treatment group underlies the increased radiosensitivity of MCF-7 cells. Further futures studies are remained elusive

  12. Vitrification of Bovine Oocytes

    Anchamparuthy, Vahida Muhammed Ismail

    2007-01-01

    Cryopreservation of oocytes is a challenge. Studies were conducted to vitrify mouse zygotes and cumulus-intact bovine oocytes from follicles of different diameters, small (â ¤ 4 mm) and medium (4 to 10 mm), using nylon mesh. The specific goals were to assess changes in apoptotic gene expression (Fas-FasL, Bax, Bcl-2, and survivin) in conjunction with terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and caspase assays. Mouse zygotes were exposed to increasing concentrations...

  13. Statins, Bcl-2 and Apoptosis: Cell Death or Cell Protection?

    Wood, W. Gibson; Igbavboa, Urule; Muller, Walter E.; Gunter P. Eckert

    2013-01-01

    Statins have proven their effectiveness in the treatment of cardiovascular disease. This class of drugs has also attracted attention as a potential treatment for dissimilar diseases such as certain types of cancers and neurodegenerative diseases. What appears to be a contradiction is that in the case of cancer, it has been suggested that statins increase apoptosis and alter levels of Bcl-2 family members (e.g., reduce Bcl-2 and increase Bax) whereas, studies mainly using non-cancerous cells r...

  14. Accelerated fibrosis and apoptosis with ageing and in atrial fibrillation: Adaptive responses with maladaptive consequences.

    Xu, Guo-Jun; Gan, Tian-Yi; Tang, Bao-Peng; Chen, Zu-Heng; Mahemuti, Ailiman; Jiang, Tao; Song, Jian-Guo; Guo, Xia; Li, Yao-Dong; Miao, Hai-Jun; Zhou, Xian-Hui; Zhang, Yu; Li, Jin-Xin

    2013-03-01

    The aim of this study was to investigate whether abnormal expression of matrix metalloproteinase (MMP)-9/tissue inhibitors of MMPs (TIMP)-1 and B cell lymphoma 2 (BCL-2)/BCL-2-associated X protein (BAX) are correlated with the characteristic accelerated fibrosis and apoptosis during ageing and in atrial fibrillation (AF). Four groups of dogs were studied: adult dogs in sinus rhythm (SR), aged dogs in SR, adult dogs with AF induced by rapid atrial pacing and aged dogs with AF induced by rapid atrial pacing. The mRNA and protein expression levels of the target gene in the left atrium were measured by quantitative reverse transcription-polymerase chain reaction (RT-PCR) and western blot analysis. Pathohistological and ultrastructural changes were assessed by light and electron microscopy. The apoptotic indices of myocytes were detected by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate (dUTP) nick end labeling (TUNEL). The mRNA and protein expression levels of MMP-9 and BAX and those of TIMP-1 and BCL-2 were significantly upregulated and down-regulated, respectively, in the aged groups compared with the adult groups. Compared with the control groups, the adult and aged groups with AF exhibited significantly increased mRNA and protein expression levels of MMP-9 and BAX and decreased expression levels of TIMP-1 and BCL-2. Samples of atrial tissue demonstrated abnormal pathohistological and ultrastructural changes, accelerated fibrosis and apoptosis. MMP-9/TIMP-1 and BCL-2/BAX hold potential for use as substrates conducive to AF and their abnormal expression plays a major role in structural remodeling of the atrium. PMID:23403858

  15. Seabuckthorn leaf extract (SBL-1) counters radiation damage by regulating time kinetics of apoptosis in jejunal crypts in total body 60Co-gamma-irradiated mice

    The radioprotective properties of plant Hippophae rhamnoides L. (common name Seabuckthorn, family Eleagnaceae) were reported and treatment with SBL-1 (herbal preparation from Seabuckthorn leaves), before whole body exposure to 60Co-gamma-rays (10 Gy), rendered >90% survivors in mice population, while 100% mortality was observed in non-SBL-1 treated, 60Co-gamma-irradiated (10 Gy) controls. Purpose of this study was to investigate the early as well as late modifying effects of SBL-1 on radiation induced apoptosis in jejunal crypts and m-RNA levels and protein levels of Bcl2 and Bax. A 30 day study was performed with 8-9 weeks old inbred male Swiss albino Strain 'A mice. Histology study was performed with jejunum to record the time dependent changes in the number of apoptotic cells in Crypts of Lieberkuhn; quantitative reverse polymerase chain reaction was performed to record the time kinetic of changes in m-RNA levels of BcI-2 and Bax genes. The changes in BcI-2 and Bax proteins were also recorded by western blotting. One time administration of SBL-1, prior to lethal whole body irradiation (10 Gy), significantly (p< 0.05) countered the radiation induced increases in cryptal apoptotic cells, Bax levels, and decrease in BcI-2 in a time dependent manner from 24 h till day 30. This study demonstrated that one of the underlying mechanisms of SBL-1 for countering radiation induced GI syndrome was by altering the time kinetics of apoptosis in cryptal cells; besides reducing the early damage. (author)

  16. A Feedback Mechanism to Control Apoptosis Occurs in the Digestive Gland of the Oyster Crassostrea gigas Exposed to the Paralytic Shellfish Toxins Producer Alexandrium catenella

    Jean-Luc Rolland; Walid Medhioub; Agnes Vergnes; Celina Abi-khalil; Véronique Savar; Eric Abadie; Estelle Masseret; Zouher Amzil; Mohamed Laabir

    2014-01-01

    To better understand the effect of Paralytic Shellfish Toxins (PSTs) accumulation in the digestive gland of the Pacific oyster, Crassostrea gigas, we experimentally exposed individual oysters for 48 h to a PSTs producer, the dinoflagellate Alexandrium catenella. In comparison to the effect of the non-toxic Alexandrium tamarense, on the eight apoptotic related genes tested, Bax and BI.1 were significantly upregulated in oysters exposed 48 h to A. catenella. Among the five detoxification relate...

  17. Intrinsic and extrinsic pathway signaling during neuronal apoptosis

    Putcha, Girish V.; Harris, Charles A; Moulder, Krista L.; Easton, Rachael M.; Thompson, Craig B.; Johnson, Eugene M.

    2002-01-01

    Trophic factor deprivation (TFD)-induced apoptosis in sympathetic neurons requires macromolecular synthesis–dependent BAX translocation, cytochrome c (cyt c) release, and caspase activation. Here, we report the contributions of other intrinsic and extrinsic pathway signals to these processes. Sympathetic neurons expressed all antiapoptotic BCL-2 proteins examined, yet expressed only certain BH3-only and multidomain proapoptotic BCL-2 family members. All coexpressed proapoptotic proteins did n...

  18. Chemical composition and deformation-induced stresses in ferroelectric films of barium-strontium titanate

    Influence of the ratio of cationic components and inner deformation-induced stresses on critical temperature (Tc) and dielectric characteristics of ferroelectric films BaxSr1-xTiO3 grown on α-Al2O3 [1012] and LaAlO3 substrates was studied. Diagnosis by means of ion backscattering permitted ascertaining the deficiency of barium in the films near the surface layer, as well as differences in their structural quality

  19. Methotrexate Promotes Platelet Apoptosis via JNK-Mediated Mitochondrial Damage: Alleviation by N-Acetylcysteine and N-Acetylcysteine Amide

    Manoj Paul; Mahadevappa Hemshekhar; Thushara, Ram M.; Mahalingam S Sundaram; NaveenKumar, Somanathapura K.; Shivanna Naveen; Sannaningaiah Devaraja; Kumar Somyajit; Robert West; Basappa,; Nayaka, Siddaiah C.; Zakai, Uzma I.; Ganesh Nagaraju; Rangappa, Kanchugarakoppal S.; Kempaiah Kemparaju

    2015-01-01

    Thrombocytopenia in methotrexate (MTX)-treated cancer and rheumatoid arthritis (RA) patients connotes the interference of MTX with platelets. Hence, it seemed appealing to appraise the effect of MTX on platelets. Thereby, the mechanism of action of MTX on platelets was dissected. MTX (10 μM) induced activation of pro-apoptotic proteins Bid, Bax and Bad through JNK phosphorylation leading to ΔΨm dissipation, cytochrome c release and caspase activation, culminating in apoptosis. The use of spec...

  20. Molecular characterization of diarrheagenic Escherichia coli strains from stools samples and food products in Colombia

    Rúgeles, Laura Cristina; Bai, Jing; Martínez, Aída Juliana; Vanegas, María Consuelo; Gómez-Duarte, Oscar Gilberto

    2010-01-01

    The prevalence of diarrheagenic E. coli in childhood diarrhea and the role of contaminated food products in disease transmission in Colombia are largely unknown. The aim of this study is to identify E. coli pathotypes, including E. coli O157:H7, from 108 stool samples from children with acute diarrhea, 38 meat samples and 38 vegetable samples. Multiplex PCR and Bax Dupont systems were used for E. coli pathotype detection. Eighteen (9.8%) E. coli diarrheagenic pathotypes were detected among al...

  1. PI3K inhibitors prime neuroblastoma cells for chemotherapy by shifting the balance towards pro-apoptotic Bcl-2 proteins and enhanced mitochondrial apoptosis.

    Bender, A; Opel, D; Naumann, I; Kappler, R; Friedman, L; von Schweinitz, D; Debatin, K-M; Fulda, S

    2011-01-27

    We recently identified activation of phosphatidylinositol 3'-kinase (PI3K)/Akt as a novel predictor of poor outcome in neuroblastoma. Here, we investigated the effect of small-molecule PI3K inhibitors on chemosensitivity. We provide first evidence that PI3K inhibitors, for example PI103, synergize with various chemotherapeutics (Doxorubicin, Etoposide, Topotecan, Cisplatin, Vincristine and Taxol) to trigger apoptosis in neuroblastoma cells (combination index: high synergy). Mechanistic studies reveal that PI103 cooperates with Doxorubicin to reduce Mcl-1 expression and Bim(EL) phosphorylation and to upregulate Noxa and Bim(EL) levels. This shifted ratio of pro- and antiapoptotic Bcl-2 proteins results in increased Bax/Bak conformational change, loss of mitochondrial membrane potential, cytochrome c release, caspase activation and caspase-dependent apoptosis. Although Mcl-1 knockdown enhances Doxorubicin- and PI103-induced apoptosis, silencing of Noxa, Bax/Bak or p53 reduces apoptosis, underscoring the functional relevance of the Doxorubicin- and PI103-mediated modulation of these proteins for chemosensitization. Bcl-2 overexpression inhibits Bax activation, mitochondrial perturbations, cleavage of caspases and Bid, and apoptosis, confirming the central role of the mitochondrial pathway for chemosensitization. Interestingly, the broad-range caspase inhibitor zVAD.fmk does not interfere with Bax activation or mitochondrial outer membrane permeabilization, whereas it blocks caspase activation and apoptosis, thus placing mitochondrial events upstream of caspase activation. Importantly, PI103 and Doxorubicin cooperate to induce apoptosis and to suppress tumor growth in patients' derived primary neuroblastoma cells and in an in vivo neuroblastoma model, underlining the clinical relevance of the results. Thus, targeting PI3K presents a novel and promising strategy to sensitize neuroblastoma cells for chemotherapy-induced apoptosis, which has important implications for the

  2. Functional significance of metastasis-inducing S100A4(Mts1) in tumor-stroma interplay

    Schmidt-Hansen, Birgitte; Klingelhöfer, Jörg; Grum-Schwensen, Birgitte;

    2004-01-01

    , extracellular S100A4 down-regulates the pro-apoptotic bax and the angiogenesis inhibitor thrombospondin-1 genes. For the first time, we demonstrate here that the S100A4 protein added to the extracellular space strongly stimulates proteolytic activity of VMR cells. This activity most probably is associated with...... in the tumor-stroma environment. S100A4 released either by tumor or stroma cells triggers pro-metastatic cascades in tumor cells....

  3. The different biological effects of single, fractioned and continuous low dose rate radiation on CL187 colorectal cancer cell line

    Objective: To investigate the effect and underlying mechanism of single, fractioned and continuous low dose rate radiation on CL187 colorectal cancer cell line. Methods: CL187 cells were exposed to 6 MV X-rays at a high dose rate of 4 Gy/min and 125I seed at a low dose rate of 2.77 cGy/h with three groups:single dose radiation group (SDR), fractioned dose radiation group (FDR) by 2 Gy/f, and continuous low dose rate radiation group (CLDR). The radiation doses were 0, 2, 4 and 8 Gy. Total cell number and cell viability were determined by trypan blue. Clone forming assay was used to evaluate the cell proliferation ability. The percentage of apoptosis cells was analyzed by flow cytometry. Western blot was used to detect the protein expression levels of PHLPP2, PTEN and Bax. Results: Compared with SDR and FDR groups, the total cell number and survival fraction of CLDR group decreased. The relative biological effect (RBE) for 125I seeds compared with 6 MV X-rays was 1.41. The percentage of apoptosis cells of CLDR group was significantly increased (t=-15.08, -11.99, P<0.05). The expression level of Bax increased in CLDR group, while no obvious changes were observed on PHLPP2 and PTEN among three groups. Conclusions: The expression level of PHLPP2 increases in SDR, FDR and CLDR group, while it seems that it was not influenced by dose rate. The expression level of Bax increased in three groups, while more colorectal CL187 cells in CLDR group may be killed due to the increase of Bax expression. (authors)

  4. Human neuronal apoptosis secondary to traumatic brain injury and the regulative role of apoptosis-related genes

    杨树源; 雪亮

    2004-01-01

    Objective: To observe human neuronal apoptosis secondary to traumatic brain injury, and to elucidate its regulative mechanism and the change of expression of apoptosis-related genes.Methods: Specimens of brain were collected from cases of traumatic brain injury in humans. The histological and cellular morphology was examined by light and electron microscopy. The extent of DNA injury to cortical neurons was detected by using TUNEL. By in situ hybridisation and immunohistochemistry the mRNA changes and protein expression of Bcl-2, Bax, p53, and caspase 3 p20 subunit were observed.Results: Apoptotic neurons appeared following traumatic brain injury, peaked at 24 hours and lasted for 7 days. In normal brain tissue activated caspase 3 was rare,but a short time after trauma it became activated. The activity peaked at 20-28 hours and remained higher than normal for 5-7 days. There was no expression of Bcl-2 mRNA and Bcl-2 protein in normal brain tissue but 8 hours after injury their expression became evident and then increased, peaked at 2-3 days and remained higher than normal for 5-7 days. The primary expression of Bax-mRNA and Bax protein was high in normal brain tissue. At 20-28 hours they increased and remained high for 2-3 days; on the 7th days they returned to a normal level. In normal brain tissue, p53mRNA and P53 were minimally expressed.Increased expression was detected at the 8th hour, and decreased at 20-28 hours but still remained higher than normal on the 5th day.Conclusions: Following traumatic injury to the human brain, apoptotic neurons appear around the focus of trauma. The mRNA and protein expression of Bcl-2, Bax and p53 and the activity of caspase 3 enzyme are increased.

  5. Spinal cord decompression reduces rat neural cell apoptosis secondary to spinal cord injury*

    Xu, Kan; Chen, Qi-xin; Li, Fang-cai; Chen, Wei-Shan; Lin, Min; Wu, Qiong-hua

    2009-01-01

    Objective: To determine whether spinal cord decompression plays a role in neural cell apoptosis after spinal cord injury. Study design: We used an animal model of compressive spinal cord injury with incomplete paraparesis to evaluate neural cell apoptosis after decompression. Apoptosis and cellular damage were assessed by staining with terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate nick-end labelling (TUNEL) and immunostaining for caspase-3, Bcl-2 and Bax. Meth...

  6. The Prognostic Significance of Apoptosis-Related Biological Markers in Chinese Gastric Cancer Patients

    Liu, Xiaowen; Cai, Hong; Huang, Hua; Long, Ziwen; Shi, Yingqiang; Wang, Yanong

    2011-01-01

    Background and Objective The prognosis varied among the patients with the same stage, therefore there was a need for new prognostic and predictive factors. The aim of this study was to evaluate the relationship of apoptosis-related biological markers such as p53, bcl-2, bax, and c-myc, and clinicopathological features and their prognostic value. Methods From 1996 to 2007, 4426 patients had undergone curative D2 gastrectomy for gastric cancer at Fudan University Shanghai Cancer Center. Among 5...

  7. Sex-related resistance to myocardial ischemia-reperfusion injury is associated with high constitutive ARC expression

    Bouma, Wobbe; Noma, Mio; Kanemoto, Shinya; Matsubara, Muneaki; Leshnower, Bradley G.; Hinmon, Robin; Gorman, Joseph H.; Gorman, Robert C

    2010-01-01

    The female sex has been associated with improved myocardial salvage after ischemia and reperfusion (I/R). Estrogen, specifically 17β-estradiol, has been demonstrated to mediate this phenomenon by limiting cardiomyocyte apoptosis. We sought to quantitatively assess the effect of sex, ovarian hormone loss, and I/R on myocardial Bax, Bcl-2, and apoptosis repressor with caspase recruitment domain (ARC) expression. Male (n = 48), female (n = 26), and oophorectomized female (n = 20) rabbits underwe...

  8. Pro-apoptotic and anti-inflammatory potential of andrographolide during 7,12-dimethylbenz[a]anthracene induced hamster buccal pouch carcinogenesis

    Karthikeyan Sekar; Baskaran Nagarethinam; Arjun Kumar Singh; Manoharan Shanmugam

    2012-01-01

    Objective: Aim of the present study was to investigate the modulating effect of andrographolide on apoptotic and inflammatory markers during 7,12-dimethyl-benz[a]anthracene (DMBA) induced hamster buccal pouch carcinogenesis. Methods: Oral tumors were developed in the buccal pouch of golden Syrian hamsters by painting with 0.5% DMBA in liquid paraffin three times a week for 14 weeks. The expression pattern of molecular markers were assayed using immunohistochemistry (p53, Bcl-2 and Bax), ELISA...

  9. Mechanical stretch induces mitochondria-dependent apoptosis in neonatal rat cardiomyocytes and G2/M accumulation in cardiac fibroblasts

    Xu Dong LIAO; Xiao Hui WANG; Hai Jing JIN; Lan Ying CHEN; Quan CHEN

    2004-01-01

    Heart remodeling is associated with the loss of cardiomyocytes and increase of fibrous tissue owing to abnormal mechanical load in a number of heart disease conditions. In present study,a well-described in vitro sustained stretch model was employed to study mechanical stretch-induced responses in both neonatal cardiomyocytes and cardiac fibroblasts. Cardiomyocytes,but not cardiac fibroblasts,underwent mitochondria-dependent apoptosis as evidenced by cytochrome c (cyto c) and Smac/DIABLO release from mitochondria into cytosol accompanied by mitochondrial membrane potential (△ψm) reduction,indicative of mitochondrial permeability transition pore (PTP)opening. Cyclosporin A,an inhibitor of PTP,inhibited stretch-induced cyto c release,△ψm reduction and apoptosis,suggesting an important role of mitochondrial PTP in stretch-induced apoptosis. The stretch also resulted in increased expression of the pro-apoptotic Bcl-2 family proteins,including Bax and Bad,in cardiomyocytes,but not in fibroblasts. Bax was accumulated in mitochondria following stretch. Cell permeable Bid-BH3 peptide could induce and facilitate stretch-induced apoptosis and △ψm reduction in cardiomyocytes. These results suggest that Bcl-2 family proteins play an important role in coupling stretch signaling to mitochondrial death machinery,probably by targeting to PTP. Interestingly,the levels of p53 were increased at 12 h after stretch although we observed that Bax upregulation and apoptosis occurred as early as 1 h. Adenovirus delivered dominant negative p53 blocked Bax upregulation in cardiomyocytes but showed partial effect on preventing stretch-induced apoptosis,suggesting that p53 was only partially involved in mediating stretch-induced apoptosis. Furthermore,we showed that p21 was upregulated and cyclin B l was downregulated only in cardiac fibroblasts,which may be associated with G2/M accumulation in response to mechanical stretch.

  10. Mitochondrial outer-membrane permeabilization and remodelling in apoptosis

    Jourdain, Alexis; Martinou, Jean-Claude

    2009-01-01

    Many human pathologies are associated with defects in mitochondria such as diabetes, neurodegenerative diseases or cancer. This tiny organelle is involved in a plethora of processes in mammalian cells, including energy production, lipid metabolism and cell death. In the so-called intrinsic apoptotic pathway, the outer mitochondrial membrane (MOM) is premeabilized by the pro-apoptotic Bcl-2 members Bax and Bak, allowing the release of apoptogenic factors such as cytochrome c from the inter-mem...

  11. Raman scattering and luminescence of high-Tc superconducting oxides

    Raman and luminescence spectra of high-Tc superconducting oxides are summarized, mainly YBa2Cu3O7-σ and partly La2-xBaxCuO4-σ. In raman spectra we succeeded to distinguish electron scattering to define the energy gap Δ in the superconducting state. The luminescence spectra are due to the emission of oxygen and interaction with conduction electrons. 70 refs.; 13 figs

  12. Rational Development of a Cytotoxic Peptide to Trigger Cell Death

    Boohaker, Rebecca J.; Zhang, Ge; Lee, Michael W; Nemec, Kathleen N.; Santra, Santimukul; Perez, J Manuel; Khaled, Annette R.

    2012-01-01

    Defects in the apoptotic machinery can contribute to tumor formation and resistance to treatment, creating a need to identify new agents that kill cancer cells by alternative mechanisms. To this end, we examined the cytotoxic properties of a novel peptide, CT20p, derived from the C-terminal, alpha-9 helix of Bax, an amphipathic domain with putative membrane binding properties. Like many anti-microbial peptides, CT20p contains clusters of hydrophobic and cationic residues that could enable the...

  13. A surface groove essential for viral bcl-2 function during chronic infection in vivo.

    2005-09-01

    Full Text Available Antiapoptotic Bcl-2 family proteins inhibit apoptosis in cultured cells by binding BH3 domains of proapoptotic Bcl-2 family members via a hydrophobic BH3 binding groove on the protein surface. We investigated the physiological importance of the BH3 binding groove of an antiapoptotic Bcl-2 protein in mammals in vivo by analyzing a viral Bcl-2 family protein. We show that the gamma-herpesvirus 68 (gammaHV68 Bcl-2 family protein (gammaHV68 v-Bcl-2, which is known to inhibit apoptosis in cultured cells, inhibits both apoptosis in primary lymphocytes and Bax toxicity in yeast. Nuclear magnetic resonance determination of the gammaHV68 v-Bcl-2 structure revealed a BH3 binding groove that binds BH3 domain peptides from proapoptotic Bcl-2 family members Bax and Bak via a molecular mechanism shared with host Bcl-2 family proteins, involving a conserved arginine in the BH3 peptide binding groove. Mutations of this conserved arginine and two adjacent amino acids to alanine (SGR to AAA within the BH3 binding groove resulted in a properly folded protein that lacked the capacity of the wild-type gammaHV68 v-Bcl-2 to bind Bax BH3 peptide and to block Bax toxicity in yeast. We tested the physiological importance of this v-Bcl-2 domain during viral infection by engineering viral mutants encoding a v-Bcl-2 containing the SGR to AAA mutation. This mutation resulted in a virus defective for both efficient reactivation of gammaHV68 from latency and efficient persistent gammaHV68 replication. These studies demonstrate an essential functional role for amino acids in the BH3 peptide binding groove of a viral Bcl-2 family member during chronic infection.

  14. The influence of Flt3 ligand on proliferation of irradiated microvascular endothelial cells

    Objective: To investigate the influence of FL on proliferation of irradiated microvascular endothelial cells. (ECVs), and possible mechanism of FL in radiation protection of ECVs. Methods: The ECVs were screened for Flt3 receptor expression by flow cytometric analysis. The proliferation of ECVs stimulated by FL was measured by the micro-culture tetrazolium assay (MTT). The apoptosis of ECVs caused by irradiation was measured with Annexin V-PI. Two apoptosis-related genes, Bcl-XL and Bax, were also analyzed by RT-PCR. Results: Flt3 receptors were expressed on the surface of ECVs. FL stimulated the proliferation of ECVs at very low concentrations (0.5-15 ng/ml) with the maximum stimulation at 15 ng/ml. A significant increase in Bax activity and a decrease in Bcl-XL activity were seen at 24 h and 48 h post-irradiation, respectively. When the culture medium with FL was added 2 h before or immediately after irradiation, the expression of Bax fell sharply at 24 h and 48 h post-irradiation. The change in Bcl-XL activity was not so marked and a mild increase in Bcl-XL expression was seen only at 48 h post-irradiation. FL inhibited the apoptosis of ECVs caused by irradiation and stimulated the proliferation of irradiated ECVs. Conclusion: FL down-regulates the expression of Bax in irradiated ECVs, and inhibits the apoptosis of the ECVs. Thus, FL may find a use in radio-protection of hematopoietic cells via protection of the microvascular endothelial cells in the bone marrow microenvironment

  15. Involvement of Sphingolipids in Ethanol Neurotoxicity in the Developing Brain

    Mitsuo Saito; Mariko Saito

    2013-01-01

    Ethanol-induced neuronal death during a sensitive period of brain development is considered one of the significant causes of fetal alcohol spectrum disorders (FASD). In rodent models, ethanol triggers robust apoptotic neurodegeneration during a period of active synaptogenesis that occurs around the first two postnatal weeks, equivalent to the third trimester in human fetuses. The ethanol-induced apoptosis is mitochondria-dependent, involving Bax and caspase-3 activation. Such apoptotic pathwa...

  16. Study of the variations in apoptotic factors in hippocampus of male rats with posttraumatic stress disorder

    Behrang Alani

    2013-01-01

    Full Text Available Background: Post-traumatic stress disorder (PTSD is a stress-related psychosomatic disorder caused by occurrence of a traumatic event and the hippocampus volume of the patients with Post-traumatic stress disorder decreased. However, the mechanisms that cause such damage are not well-understood. The aim of this study is to detect the expression of apoptosis-related Bax, Bcl-2, Caspase-3 and Insulin-like growth Factor-I proteins in the hippocampus region in the Predatory stress rats. Materials and Methods: A total of 70 male wistar rats were divided into Predatory stress groups of 1d, 2d, 3d, 7d, 14d, 30d and a normal control group (N = 10. Rats were subjected to 5 min of predatory stress and then exposed to the elevated plus-maze (EPM. Serum corticosterone and Insulin-like growth factor-1 level of Hippocampus were measured by ELISA technique. The expression of Bax, Bcl-2, and Caspase-3 were detected by western blotting. Results: Rats spent significantly more time in closed arms of the elevated plus maze (EPM than control group after exposure to stress. Serum levels of corticosterone significantly increased at 2d-3d. The expression of hippocampal IGF-1 was significantly up-regulated at 1d-2d after stress. Both Bax and the ratio of Bax/Bcl-2 significantly peaked at Predatory stress 2d-14d. Caspase3 was significantly active among 2d-30 compared to the normal control. Conclusion: The activation of caspase-3 in the stress groups indicates that apoptosis may be one of the reasons inducing hippocampus atrophy and play roles in the pathogenesis of PTSD. Increase in hippocampus levels of IGF-1 during early PTSD might be involved in the early molecular inhibitory mechanism of apoptosis in PTSD.

  17. Eurycomanone induce apoptosis in HepG2 cells via up-regulation of p53

    Zakaria Yusmazura

    2009-06-01

    Full Text Available Abstract Background Eurycomanone is a cytotoxic compound found in Eurycoma longifolia Jack. Previous studies had noted the cytotoxic effect against various cancer cell lines. The aim of this study is to investigate the cytotoxicity against human hepato carcinoma cell in vitro and the mode of action. The cytotoxicity of eurycomanone was evaluated using MTT assay and the mode of cell death was detected by Hoechst 33258 nuclear staining and flow cytometry with Annexin-V/propidium iodide double staining. The protein expression Bax, Bcl-2, p53 and cytochrome C were studied by flow cytometry using a spesific antibody conjugated fluorescent dye to confirm the up-regulation of p53 and Bax in cancer cells. Results The findings suggested that eurycomanone was cytotoxic on cancerous liver cell, HepG2 and less toxic on normal cells Chang's liver and WLR-68. Furthermore, various methods proved that apoptosis was the mode of death in eurycomanone-treated HepG2 cells. The characteristics of apoptosis including chromatin condensation, DNA fragmentation and apoptotic bodies were found following eurycomanone treatment. This study also found that apoptotic process triggered by eurycomanone involved the up-regulation of p53 tumor suppressor protein. The up-regulation of p53 was followed by the increasing of pro-apoptotic Bax and decreasing of anti-apoptotic Bcl-2. The increased of cytochrome C levels in cytosol also results in induction of apoptosis. Conclusion The data suggest that eurycomanone was cytotoxic on HepG2 cells by inducing apoptosis through the up-regulation of p53 and Bax, and down-regulation of Bcl-2.

  18. Genetic deficiency of adiponectin protects against acute kidney injury

    Jin, Xiaogao; Chen, Jiyuan; Hu, Zhaoyong; Chan, Lawrence; Wang, Yanlin

    2013-01-01

    Adiponectin is a multifunctional cytokine that has a role in regulating inflammation. Here we determined if adiponectin modulates ischemic acute kidney injury. Compared with wild-type mice, adiponectin knockout mice were found to have lower serum creatinine and less tubular damage or apoptosis following ischemia/reperfusion injury. This latter process was associated with decreased Bax and reduced activation of p53 and caspase-3. Targeted disruption of adiponectin was also found to inhibit the...

  19. Effect of small dose of radiation on induction of apoptosis in murine tumors

    To investigate the presence of adaptive response by low dose radiation in murine tumors in relation to radiation induced apoptosis as well as related mechanism. Syngeneic murine tumors, OCa-1 and HCa-l, were given 0.05 Gy pretreatment followed by therapeutic dose of 25 Gy radiation. Induction of apoptosis was analyzed for each treatment group. Regulating molecules of apoptosis. p53, Bcl-2, Sax, Bel-X, were also analyzed by Western blotting. In 0.05 Gy pretreatment group of OCa-l, 25 Gy-induced apoptosis per 1000 cells was 229, which was estimated at 30% lower level than the expected (p<0.05). In contrast, this reduction in radiation induced apoptosis was not seen in HCa-1. In the expression of apoptosis regulating molecules, p53 increased in both tumors in response to radiation. Bcl-2 and Bax did not show significant change in both tumors however, the expression of Bcl-2 surpassed that of Bax in 0.05 Gy pretreatment group of OCa-1. Bcl-X was not expressed in OCa-1. In HCa-l, ScI-X showed increased expression even with 0.05 Gy. Adaptive response by low dose radiation is shown in one murine tumor, OCa-I, in relation to radiation induced apoptosis. Apoptosis regulating molecules including Bcl-2/Bax and Bcl-X, appear to related. This study shows an evidence that adaptive response is present, but not a generalized phenomenon in vivo

  20. Neuroprotective Effects of Dexmedetomidine Against Hypoxia-Induced Nervous System Injury are Related to Inhibition of NF-κB/COX-2 Pathways.

    Pan, Wanying; Lin, Lin; Zhang, Nan; Yuan, Fuli; Hua, Xiaoxiao; Wang, Yueting; Mo, Liqiu

    2016-10-01

    Dexmedetomidine has been reported to provide neuroprotection against hypoxia-induced damage. However, the underlying mechanisms remain unclear. We examined whether dexmedetomidine's neuroprotective effects were mediated by the NF-κB/COX-2 pathways. Adult male C57BL/6 mice were subjected to a 30-min hypoxic treatment followed by recovery to normal conditions. They received dexmedetomidine (16 or 160 μg/kg) or 25 mg/kg atipamezole, an α2-adrenoreceptor antagonist, intraperitoneally before exposure to hypoxia. The whole brain was harvested 6, 18, or 36 h after the hypoxia to determine the histopathological outcome and cleaved caspase-3, Bax/Bcl, NF-κB, and COX-2 levels. Hypoxia treatment induced significant neurotoxicity, including destruction of the tissue structure and upregulation of the protein levels of caspase-3, the ratio of Bax/Bcl-2, NF-κB, and COX-2. Dexmedetomidine pretreatment effectively improved histological outcome and restored levels of caspase-3, the Bax/Bcl-2 ratio, NF-κB, and COX-2. Atipamezole reversed the neuroprotection induced by dexmedetomidine. Neuroprotection was achieved by PDTC and NS-398, inhibitors of NF-κB and COX-2, respectively. Dexmedetomidine use before hypoxia provides neuroprotection. Inhibition of NF-κB/COX-2 pathways activation may contribute to the neuroprotection of dexmedetomidine. PMID:26683659