Preparation, Purification and Regioselective Functionalization of Protoescigenin—The Main Aglycone of Escin Complex

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Krzysztof Woźniak

2013-04-01

Full Text Available A two-step chemical process for controlled degradation of escin, affording a mixture of olean-12-ene sapogenins, was elaborated and scaled up. The main component of the mixture—protoescigenin—was isolated and purified, in the form of its corresponding monohydrate, without resource to chromatographic methods. This material was further converted into the high purity 3,24;16,22-di-O,O-isopropylidene derivative in a validated large scale laboratory process.

Isolation, structural determination, and evaluation of the biological activity of 20(S)-25-methoxyl-dammarane-3beta, 12beta, 20-triol [20(S)-25-OCH3-PPD], a novel natural product from Panax notoginseng.

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Zhao, Y; Wang, W; Han, L; Rayburn, E R; Hill, D L; Wang, H; Zhang, R

2007-01-01

Ginseng has been used extensively for medicinal purposes, with suggested utility for indications as diverse as diabetes, cardiovascular disease and cancer. Herein we report the discovery and characterization of 20(S)-25-OCH3-PPD, a ginsenoside that inhibits growth and survival of cancer cells. The novel dammarane triterpene sapogenin (C31H56O4; molecular weight 492) was isolated from the total hydrolyzed saponins extracted from the leaves of Panax notoginseng using conventional and reverse-phase silica gel chromatography. Based on physicochemical characteristics and NMR data, the compound was identified as 20(S)-25-OCH3-PPD. The biological activities of 20(S)-25-OCH3-PPD and its known analogs, 20(S)-PPD and Rg3, were evaluated in 12 human cancer cell lines. In all cell lines, the order of cytotoxicity of the test compounds was 20(S)-25-OCH3-PPD > 20(S)-PPD > Rg3. 20(S)-25-OCH3-PPD also induced apoptosis and cell cycle arrest in the G1 phase, and inhibited proliferation in breast cancer cell lines, demonstrating its potent biological effects. In regard to cytotoxicity, the IC50 values of 20(S)-25-OCH3-PPD for most cell lines were in the lower microM range, a 5-15-fold greater cytotoxicity relative to 20(S)-PPD and a 10-100-fold increase over Rg3. These findings suggest a structure-activity relationship among dammarane-type sapogenins. The data presented here may provide a basis for the future development of 20(S)-25-OCH3-PPD as a novel anti-cancer agent.

Saponin Isolation as Main Ingredients of Insecticide and Collagen Type I From Crown of Thorn-Starfish (Acanthaster planci)

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Wijanarko, Anondho; Januardi Ginting, Mikael; Sahlan, Muhamad; Krisanta Endah Savitri, Imelda; Florensia, Yunita; Sudiarta, Maria Regina; Pastika, Satria; Rafiki, Fakhri; Hermansyah, Heri

2017-10-01

The outbreaks of crown of thorns starfish (Acanthaster planci) resulted in the severe destruction of coral reefs in a large number of Indonesia’s marine ecosystem, especially in the western part. At the moment, control efforts are proven to be ineffective because of its high cost and labor intensive. Recent research found that A. planci contain saponins that act as cytotoxic compound and can be used as an environment-friendly insecticide to eradicate Kalotermitidae pest. Saponins extracted by maceration using ethanol 96.0% with a total yield of saponins 9.04% and 4.66% for two test. Purification of saponin was achieved by utilization of activated carbon with a mass of carbon:volume sample 1:2 (w/v) and stirred for 20 minutes. Sapogenin can be isolated by hydrolyzing using hydrochloric acid, and thus 168.4 mg sapogenin is obtained. In addition to saponins, A. planci also contains collagen Type I. Collagen isolation by multistage extraction began with extracting the collagen with alkaline solvent, with water, NaOH 0.1 M, and Ca(OH)2 0.2 M as the solvent variations. The second step is acid-enzymatic extraction by pepsin digestion in 0.5 M acetic acid. Collagen extract will be further purified by salting out and dialysis method to obtain pure collagen yield called Pepsin Solubilized Collagens (PSC). Characterization of PSC consists of quantitative and qualitative analysis such as Lowry method, gel electrophoresis, UV spectroscopy, amino acid composition analysis, and Scanning Electron Microscopy (SEM). The result shows Ca(OH)2 0.2 M as the best extraction solvent with 2.26% yield of PSC.

Identification and genome organization of saponin pathway genes from a wild crucifer, and their use for transient production of saponins in Nicotiana benthamiana.

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Khakimov, Bekzod; Kuzina, Vera; Erthmann, Pernille Ø; Fukushima, Ery Odette; Augustin, Jörg M; Olsen, Carl Erik; Scholtalbers, Jelle; Volpin, Hanne; Andersen, Sven Bode; Hauser, Thure P; Muranaka, Toshiya; Bak, Søren

2015-11-01

The ability to evolve novel metabolites has been instrumental for the defence of plants against antagonists. A few species in the Barbarea genus are the only crucifers known to produce saponins, some of which make plants resistant to specialist herbivores, like Plutella xylostella, the diamondback moth. Genetic mapping in Barbarea vulgaris revealed that genes for saponin biosynthesis are not clustered but are located in different linkage groups. Using co-location with quantitative trait loci (QTLs) for resistance, transcriptome and genome sequences, we identified two 2,3-oxidosqualene cyclases that form the major triterpenoid backbones. LUP2 mainly produces lupeol, and is preferentially expressed in insect-susceptible B. vulgaris plants, whereas LUP5 produces β-amyrin and α-amyrin, and is preferentially expressed in resistant plants; β-amyrin is the backbone for the resistance-conferring saponins in Barbarea. Two loci for cytochromes P450, predicted to add functional groups to the saponin backbone, were identified: CYP72As co-localized with insect resistance, whereas CYP716As did not. When B. vulgaris sapogenin biosynthesis genes were transiently expressed by CPMV-HT technology in Nicotiana benthamiana, high levels of hydroxylated and carboxylated triterpenoid structures accumulated, including oleanolic acid, which is a precursor of the major resistance-conferring saponins. When the B. vulgaris gene for sapogenin 3-O-glucosylation was co-expressed, the insect deterrent 3-O-oleanolic acid monoglucoside accumulated, as well as triterpene structures with up to six hexoses, demonstrating that N. benthamiana further decorates the monoglucosides. We argue that saponin biosynthesis in the Barbarea genus evolved by a neofunctionalized glucosyl transferase, whereas the difference between resistant and susceptible B. vulgaris chemotypes evolved by different expression of oxidosqualene cyclases (OSCs). © 2015 The Authors The Plant Journal © 2015 John Wiley & Sons

Break-through in seed fertility of autotetraploid Trigonella foenum graecum

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Raghuvanshi, S S; Agrawal, Poonam; Singh, Jaya; Singh, Anil K [Plant Genetics Unit, Botany Department, Lucknow University, Lucknow (India)

1990-01-01

Full text: Fenugreek is an important legume crop in tropical and subtropical zones of India, used as green vegetable, cattle fodder, food flavourant and for pharmaceutical purposes. The seeds are a source of diosgenin, a steroid sapogenin used for synthesis of steroidal drugs like corticosteroide sex hormone and oral contraceptives. Seeds valuing 15-20 million rupees are exported annually. Polyploids are vegetatively superior but have poorer seed fertility. Cross breeding among locally collected germplasm and autopolyploidisation did not give satisfactory results. Mutation induction using gamma rays or EMS alone and in combination was undertaken to broaden the variability for increased grain yield. In M{sub 3}, mutants were selected with up to 143 pods per plant, compared to 80 in the diploid progenitors and up to 123 In advanced hybrids. 11 promising high yielding lines will be further evaluated. (author)

Break-through in seed fertility of autotetraploid Trigonella foenum graecum

International Nuclear Information System (INIS)

Raghuvanshi, S.S.; Agrawal, Poonam; Singh, Jaya; Singh, Anil K.

1990-01-01

Full text: Fenugreek is an important legume crop in tropical and subtropical zones of India, used as green vegetable, cattle fodder, food flavourant and for pharmaceutical purposes. The seeds are a source of diosgenin, a steroid sapogenin used for synthesis of steroidal drugs like corticosteroide sex hormone and oral contraceptives. Seeds valuing 15-20 million rupees are exported annually. Polyploids are vegetatively superior but have poorer seed fertility. Cross breeding among locally collected germplasm and autopolyploidisation did not give satisfactory results. Mutation induction using gamma rays or EMS alone and in combination was undertaken to broaden the variability for increased grain yield. In M 3 , mutants were selected with up to 143 pods per plant, compared to 80 in the diploid progenitors and up to 123 In advanced hybrids. 11 promising high yielding lines will be further evaluated. (author)

Synthesis and in vitro anticancer activity of 23(23′)E-benzylidenespirostanols derived from steroid sapogenins

Czech Academy of Sciences Publication Activity Database

Ramos-Enríquez, M. A.; Vargas-Romero, K.; Rárová, L.; Strnad, Miroslav; Iglesias-Arteaga, M. A.

2017-01-01

Roč. 128, DEC (2017), s. 85-88 ISSN 0039-128X R&D Projects: GA ČR GA17-14007S Institutional support: RVO:61389030 Keywords : Aldol condensation * Benzylidenespirostanols * Cancer cell lines * Cytotoxicity * Fibroblasts Subject RIV: FD - Oncology ; Hematology OBOR OECD: Oncology Impact factor: 2.282, year: 2016

Inhibitory Effect of Triterpenoids from Panax ginseng on Coagulation Factor X

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Lingxin Xiong

2017-04-01

Full Text Available Enzymes involved in the coagulation process have received great attention as potential targets for the development of oral anti-coagulants. Among these enzymes, coagulation factor Xa (FXa has remained the center of attention in the last decade. In this study, 16 ginsenosides and two sapogenins were isolated, identified and quantified. To determine the inhibitory potential on FXa, the chromogenic substrates method was used. The assay suggested that compounds 5, 13 and 18 were mainly responsible for the anti-coagulant effect. Furthermore, these three compounds also possessed high thrombin selectivity in the thrombin inhibition assay. Furthermore, Glide XP from Schrödinger was employed for molecular docking to clarify the interaction between the bioactive compounds and FXa. Therefore, the chemical and biological results indicate that compounds 5 (ginsenoside Rg2, 13 (ginsenoside Rg3 and 18 (protopanaxtriol, PPT are potential natural inhibitors against FXa.

Medicinal significance, pharmacological activities, and analytical aspects of solasodine: A concise report of current scientific literature

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Kanika Patel

2013-01-01

Full Text Available Alkaloids are well known phytoconstituents for their diverse pharmacological properties. Alkaloids are found in all plant parts like roots, stems, leaves, flowers, fruits and seeds. Solasodine occurs as an aglycone part of glycoalkloids, which is a nitrogen analogue to sapogenins. Solanaceae family comprises of a number of plants with variety of natural products of medicinal significance mainly steroidal lactones, glycosides, alkaloids and flavanoids. It is a steroidal alkaloid based on a C27 cholestane skeleton. Literature survey reveals that solasodine has diuretic, anticancer, antifungal, cardiotonic, antispermatogenetic, antiandrogenic, immunomodulatory, antipyretic and various effects on central nervous system. Isolation and quantitative determination was achieved by several analytical techniques. Present review highlights the pharmacological activity of solasodine, with its analytical and tissue culture techniques, which may be helpful to the researchers to develop new molecules for the treatment of various disorders in the future.

Nutrient composition of Dacryodes edulis seed and seed coat mixture

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C.U. OGUNKA-NNOKA

2017-07-01

Full Text Available This study investigated the nutrient composition of D. edulis seed and seed coat mixture. Qualitative and quantitative phytochemicals, proximate, and vitamin compositions were evaluated using standard methods. Saponins were very high, alkaloids, flavonoids, and tannins were high, while terpenoids were low, and glycosides, aldehydes, and steroids were absent. The quantitative phytochemical determination followed the order; saponin > kaempferol > rutin > catechin > tannin > sapogenin > lunamarine > phenol > ribalinidine > anthocyanin > oxalate > phytate. For the proximate composition, carbohydrates had the highest concentration, followed by lipids and fibre, while, protein concentration was the lowest. Vitamin E (5.42 mg/100g, vitamin C (3.24 mg/100g, and vitamin A (2.84 mg/100g were the highest occurring constituent vitamins while vitamin B12 (0.035 mg/100g and vitamin B2 (0.075 mg/100g were the least occurring vitamins. This study has shown the rich phytochemical composition of D. edulis seed and seed coat mixture while showing deficiencies in proteins, distinct vitamins, and ash contents.

Steroidal Saponins

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Sahu, N. P.; Banerjee, S.; Mondal, N. B.; Mandal, D.

The medicinal activities of plants are generally due to the secondary metabolites (1) which often occur as glycosides of steroids, terpenoids, phenols etc. Saponins are a group of naturally occurring plant glycosides, characterized by their strong foam-forming properties in aqueous solution. The cardiac glycosides also possess this, property but are classified separately because of their specific biological activity. Unlike the cardiac glycosides, saponins generally do not affect the heart. These are classified as steroid or triterpenoid saponins depending on the nature of the aglycone. Steroidal glycosides are naturally occurring sugar conjugates of C27 steroidal compounds. The aglycone of a steroid saponin is usually a spirostanol or a furostanol. The glycone parts of these compounds are mostly oligosaccharides, arranged either in a linear or branched fashion, attached to hydroxyl groups through an acetal linkage (2, 3). Another class of saponins, the basic steroid saponins, contain nitrogen analogues of steroid sapogenins as aglycones.

Direct detection of saponins in crude extracts of soapnuts by FTIR.

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Almutairi, Meshari Saad; Ali, Muhammad

2015-01-01

Direct detection of saponins in soapnuts (Sapindus mukorossi) using Fourier transform infrared (FTIR) spectroscopy is investigated in this project. Potassium bromide powder was mixed with extracted powder of soapnuts and compressed to a thin pellet for examination process. The outcome of the FTIR spectra of saponin demonstrated characteristic triterpenoid saponin absorptions of OH, C = O, C-H, and C = C, while the glycoside linkages to the sapogenins were indicated by the absorptions of C-O. The significance of this study is that saponin absorption peaks are directly detectable in crude aqueous and 95% ethanol extracts of soapnuts powder using FTIR spectroscopy, thereby eliminating the need of further expensive and exhaustive purification steps. The extracts of soapnuts were screened for saponins along with controls by phytochemical tests, and advanced spectroscopic techniques such as ultra fast liquid chromatography and ultra performance liquid chromatography quadrupole-time of flight-mass spectrometry were also implemented to validate the saponins.

Effect of Agave americana and Agave salmiana Ripeness on Saponin Content from Aguamiel (Agave Sap).

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Leal-Díaz, Ana María; Santos-Zea, Liliana; Martínez-Escobedo, Hilda Cecilia; Guajardo-Flores, Daniel; Gutiérrez-Uribe, Janet Alejandra; Serna-Saldivar, Sergio Othón

2015-04-22

Steroidal saponins have shown beneficial health effects. Agave spp. leaves and rhizomes are sources of these compounds, but their presence has not been reported in the aguamiel. Aguamiel is the sweet edible sap from mature agave, and its quality is influenced by the plant ripening stage. The purpose of this research was to identify and quantitate saponins in aguamiel from Agave americana and Agave salmiana at two ripening stages. Saponins and sapogenins were identified with HPLC/ESI-MS/TOF and quantitated with HPLC/ELSD. Results proved the presence of saponins derived from kammogenin, manogenin, gentrogenin, and hecogenin. The saponin content in aguamiel from immature A. salmiana was 2-fold higher (478.3 protodioscin equivalents (PE) μg/g aguamiel (DM)) compared with A. americana (179.0 PE μg/g aguamiel (DM)). In both species, saponin content decreased when plants reached sexual maturity. This should be considered before evaluating the effects of Agave spp. as a source of bioactive saponins.

Comparative analysis of diosgenin in Dioscorea species and related medicinal plants by UPLC-DAD-MS.

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Yi, Tao; Fan, Lan-Lan; Chen, Hong-Li; Zhu, Guo-Yuan; Suen, Hau-Man; Tang, Yi-Na; Zhu, Lin; Chu, Chu; Zhao, Zhong-Zhen; Chen, Hu-Biao

2014-08-09

Dioscorea is a genus of flowering plants, and some Dioscorea species are known and used as a source for the steroidal sapogenin diosgenin. To screen potential resource from Dioscorea species and related medicinal plants for diosgenin extraction, a rapid method to compare the contents of diosgenin in various plants is crucial. An ultra-performance liquid chromatography (UPLC) coupled with diode array detection (DAD) and electrospray ionization mass spectrometry (ESI-MS) method was developed for identification and determination of diosgenin in various plants. A comprehensive validation of the developed method was conducted. Twenty-four batches of plant samples from four Dioscorea species, one Smilax species and two Heterosmilax species were analyzed by using the developed method.The present method presented good sensitivity, precision and accuracy. Diosgenin was found in three Dioscorea species and one Heterosmilax species, namely D. zingiberensis, D. septemloba, D. collettii and H. yunnanensis. The method is suitable for the screening of diosgenin resources from plants. D. zingiberensis is an important resource for diosgenin harvesting.

PERAN PEPOHONAN DALAM PENINGKATAN PRODUKSI TERNAK RUMINANSIA: PENDEKATAN ILMIAH

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I.M Mastika

2014-06-01

Full Text Available Trees and shrubs plantation are very potential and important in producing sustainable green biomass, and have high protein content and high digestibility.  Some of them are legume which are important in catching nitrogen from the air to enriched the soil nitrogen. Trees are also catching more sun energy radiation which is latter converted into animal feeds. Other roles of this plant is part of them such as leaves, fruits, barks and roots containing saponin/sapogenin or tannin which are useful as defaunating agent. It was well documented that rumen protozoa reduced ruminant productivity. From the reports available either in vitro or in vivo it was proved that saponin could to some extent reduced or depressed protozoa rumen population. Indonesia needs some  experts effort to identify and recording those plants that could play roles as natural defaunating agent, as well as provide green biomass for ruminant. The overall conditions above will provide an ample opportunities for research for students or experties.

Pengaruh Fermentasi Bakteri Asam Laktat Terhadap Sifat Fisikokimia Tepung Gadung Modifikasi (Dioscorea hispida

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R. Haryo Bimo Setiarto

2016-06-01

Full Text Available Yam (Dioscorea hispida is one of the tubers belonging to the family dioscoreaceae. Yam tubers not only have high content of carbohydrate, but also contain some toxic compounds such as: cyanogenic glycosides, alkaloids dioscorin, dehydrodioscorin, saponin and sapogenin. Fermentation of yam tubers can affected for physicochemical and amylography characteristics of modified yam flour and reduced toxic compounds. This study was aimed to determine effect of lactic acid bacteria (LAB fermentation on the physicochemical properties and characteristics of yam flour amilography modification. From the 15 isolates of Lactobacillus plantarum had been selected two isolate (L. plantarum B291 and B307 with the highest amylase enzyme activity to be used as starter fermentation to produce modified yam flour. Fermentation of LAB could reduce levels of HCN on modified yam flour, although decreased levels of HCN was not significant with requirements of SNI. Fermentation of LAB increased levels of protein, fat and lactic acid, but decreased pH value and carbohydrate content in modified yam flour. Based on results of amylography analysis, control of yam flour with shredded without fermentation had the best gelatinization profile because it most resistant about heating.ABSTRAKGadung (Dioscorea hispida merupakan salah satu jenis umbi-umbian yang tergolong dalam family Dioscoreaceae.  Ubi gadung memiliki kandungan karbohidrat yang tinggi, akan tetapi juga mengandung beberapa senyawa racun berupa glikosida sianogenik, alkaloid dioscorin, dehydrodioscorin, saponin dan sapogenin yang berbahaya bagi kesehatan. Fermentasi umbi gadung dapat mempengaruhi sifat fisikokimia dan amilografi tepung gadung modifikasi serta menurunkan senyawa toksiknya. Penelitian ini bertujuan untuk mengetahui pengaruh bakteri asam laktat (BAL penghasil amilase terhadap sifat fisikokimia dan amilografi tepung gadung modifikasi. Sebanyak 15 isolat Lactobacillus plantarum telah diseleksi, sehingga

Diosgenin: Recent Highlights on Pharmacology and Analytical Methodology.

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Jesus, Mafalda; Martins, Ana P J; Gallardo, Eugenia; Silvestre, Samuel

2016-01-01

Diosgenin, a steroidal sapogenin, occurs abundantly in plants such as Dioscorea alata , Smilax China, and Trigonella foenum graecum . This bioactive phytochemical not only is used as an important starting material for the preparation of several steroidal drugs in the pharmaceutical industry, but has revealed also high potential and interest in the treatment of various types of disorders such as cancer, hypercholesterolemia, inflammation, and several types of infections. Due to its pharmacological and industrial importance, several extraction and analytical procedures have been developed and applied over the years to isolate, detect, and quantify diosgenin, not only in its natural sources and pharmaceutical compositions, but also in animal matrices for pharmacodynamic, pharmacokinetic, and toxicological studies. Within these, HPLC technique coupled to different detectors is the most commonly analytical procedure described for this compound. However, other alternative methods were also published. Thus, the present review aims to provide collective information on the most recent pharmacological data on diosgenin and on the most relevant analytical techniques used to isolate, detect, and quantify this compound as well.

Diosgenin: Recent Highlights on Pharmacology and Analytical Methodology

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Mafalda Jesus

2016-01-01

Full Text Available Diosgenin, a steroidal sapogenin, occurs abundantly in plants such as Dioscorea alata, Smilax China, and Trigonella foenum graecum. This bioactive phytochemical not only is used as an important starting material for the preparation of several steroidal drugs in the pharmaceutical industry, but has revealed also high potential and interest in the treatment of various types of disorders such as cancer, hypercholesterolemia, inflammation, and several types of infections. Due to its pharmacological and industrial importance, several extraction and analytical procedures have been developed and applied over the years to isolate, detect, and quantify diosgenin, not only in its natural sources and pharmaceutical compositions, but also in animal matrices for pharmacodynamic, pharmacokinetic, and toxicological studies. Within these, HPLC technique coupled to different detectors is the most commonly analytical procedure described for this compound. However, other alternative methods were also published. Thus, the present review aims to provide collective information on the most recent pharmacological data on diosgenin and on the most relevant analytical techniques used to isolate, detect, and quantify this compound as well.

New compounds from acid hydrolyzed products of the fruits of Momordica charantia L. and their inhibitory activity against protein tyrosine phosphatas 1B.

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Zeng, Ke; He, Yan-Ni; Yang, Di; Cao, Jia-Qing; Xia, Xi-Chun; Zhang, Shi-Jun; Bi, Xiu-Li; Zhao, Yu-Qing

2014-06-23

Four new cucurbitane-type triterpene sapogenins, compounds 1-4, together with other eight known compounds were isolated from the acid-hydrolyzed fruits extract of Momordica charantia L. Their chemical structures were established by NMR, mass spectrometry and X-ray crystallography. Compounds 1-7 and 9-12 were evaluated for their inhibitory activities toward protein tyrosine phosphatase 1B (PTP1B), a tyrosine phosphatase that has been implicated as a key target for therapy against type II diabetes. Compounds 1, 2, 4, 7 and 9 were shown inhibitory activities of 77%, 62%, 62% 60% and 68% against PTP1B, respectively. All of these tested compounds were exhibited higher PTP1B inhibition activities than that of the Na3VO4, a known PTP1B inhibitor used as positive control in present study. Structure activity relationship (SAR) analysis indicated that the inhibition activity of PTP1B was associated with the presence and number of -OH groups. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

PENGARUH FERMENTASI BAKTERI ASAM LAKTAT LACTOBACILLUS PLANTARUM B307 TERHADAP KADAR PROKSIMAT DAN AMILOGRAFI TEPUNG TAKA MODIFIKASI (TACCA LEONTOPETALOIDES

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Raden Haryo Bimo Setiarto

2016-04-01

Full Text Available Tacca (Tacca leontopetaloides is plant that grows in coastal areas and high salinity, especially in the south coast of West Java. Tacca tubers have high content of carbohydrate, but it also contains some toxic compounds such as: taccaline, β – sitosterol, alcohol cerylic, and steroid sapogenin that are harmful for health. Fermentation on tacca tubers can change amylograph properties and proximate levels of modified tacca flour and reduce it toxic compounds. This study aimed at determining the effects of lactic acid bacteria (LAB Lactobacillus plantarum B307 fermentation on the proximate levels and amylograph characteristics of modified tacca flour. Moisture and ash content of modified tacca flour still meet the requirements of SNI. Fermentation LAB Lactobacillus plantarum B307 led to increased levels of protein and lactic acid in the modified tacca flour, but it decreased pH value and carbohydrate content. Based on the analysis of amylograph, it can be concluded that tacca flour control without fermentation has the best gelatinization profile because it has good ability of setback viscosity.

Effect of Hecogenin on DNA instability

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Marina Sampaio Cruz

Full Text Available Hecogenin is a sapogenin found in Agave species in high quantities and is responsible for the many therapeutic effects of these medicinal plants. In addition, this compound is also widely used in the pharmaceutical industry as a precursor for the synthesis of steroidal hormones and anti-inflammatory drugs. Despite Hecogenin being widely used, little is known about its toxicological properties. Therefore, the present study aimed to investigate the cytotoxic, genotoxic and mutagenic effects of Hecogenin on HepG2 cells. Cytotoxicity was analyzed using the MTT test. Then, genotoxic and mutagenic potentials were assessed by comet assay and cytokinesis-block micronucleus assay, respectively. Cytotoxic effect was observed only when cells were exposed to concentrations of Hecogenin equal or higher than 100 μM. Although a lower concentration of Hecogenin caused DNA damage, a reduction on nuclear mutagenic markers in HepG2 cells was observed. The results indicated that Hecogenin treatment generated DNA damage, but in fact it would be repaired, avoiding dissemination of the damage throughout the cell division. Further studies need to be performed to confirm the observed protective effect of Hecogenin against genomic instability. Keywords: Hecogenin, CBMN, Genotoxicity, Comet assay

Selection of mutants of Chenopodium quinoa Willd. in the generation M2, quantification of saponins on the Islunga and Barandales varieties adapted to the Toluca Valley

International Nuclear Information System (INIS)

Fernandez V, S.O.

1996-01-01

The saponins and total lipids was measured in the variants of Barandales and Isluga of Chenopodium quinoa Willd and the mutants of both variants in the M 2 lineage. These two variants were gamma radiated at different levels: 0, 50, 100, 150, 200, 250, 300, 350, 400 and 500 Gy in the M1 generation. At the same time the mutants with favorable agronomic characteristics were selected. The samples were growth on the field (cultivated) and the seeds were recovered (harvested), measuring the emergence agronomic variables, plant height, density and seed yield. Lipids totals and saponins were measured in the seeds. The samples of radiated seeds with gamma-ray at different levels, were dried and grinded until to obtain a powder. The lipids was extracted with diethyl-ether, and in this solution was measured the total lipid content. The saponins were extracted in a later step, concentrated and precipitated. After the precipitation step the sapogenins were hydrolized and extracted with methyl alcohol. The extracts were centifugated and the solid material was dried and weighted. Finally, the measurement of the oleanolic acid was carried out by high pressure liquid chromatography. One conversion factor permits to relate this acid with the total saponin content. (Author)

Synthesis of specifically deuterium-labelled pregnanolone and pregnanediol sulphates for metabolic studies in humans.

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Baillie, T A; Sjövall, J; Herz, J E

1975-10-01

A synthesis is reported of 3beta-hydroxy-5alpha-pregnan-20-one sulphate and the disulphate and 3-monosulphate of 5alpha-pregnane-3beta,20alpha-diol, labelled specifically with deuterium in high isotopic purity for metabolic studies in humans. Base-catalyzed equilibration of 3beta-hydroxy-5alpha-25R-spirostan-12-one (hemcogenin, II) with deuterium oxide, followed by removal of the 12-keto group and degradation of the sapogenin side-chain afforded 3beta-hydroxy-5alpha-[11,11-2H2]pregn-16-en-20-one (VII). Further deuterium atoms were introduced at the 3alpha and 20beta positions by reductions with sodium borodeuteride and lithium aluminum deuteride, respectively. These reactions led to 3beta-hydroxy-5alpha-[3alpha,11,11-2H3]pregnan-20-one (X; isotopic purity 87.2%) and 5alpha-[3alpha,11,11,20beta-2H4]pregnane-3beta,20alpha-diol (XIV; isotopic purity 83.9%). The 3-sulphate of the pregnanolone and the 3,20-disulphate of the pregnanediol were prepared directly form the free alcohols, while the 3-monosulphate of the pregnanediol was obtained via 5alpha-[3alpha,11,11,20beta-2H4]pregnane-3beta,20alpha-diol 20-acetate (XVII).

Selection of mutants of Chenopodium quinoa Willd. in the generation M{sub 2}, quantification of saponins on the Islunga and Barandales varieties adapted to the Toluca Valley; Seleccion de mutantes de Chenopodium quinoa Willd. en la generacion M{sub 2}, cuantificacion de saponinas en las variedades Isluga y Barandales adaptadas al Valle de Toluca

Energy Technology Data Exchange (ETDEWEB)

Fernandez V, S O

1997-12-31

The saponins and total lipids was measured in the variants of Barandales and Isluga of Chenopodium quinoa Willd and the mutants of both variants in the M{sub 2} lineage. These two variants were gamma radiated at different levels: 0, 50, 100, 150, 200, 250, 300, 350, 400 and 500 Gy in the M1 generation. At the same time the mutants with favorable agronomic characteristics were selected. The samples were growth on the field (cultivated) and the seeds were recovered (harvested), measuring the emergence agronomic variables, plant height, density and seed yield. Lipids totals and saponins were measured in the seeds. The samples of radiated seeds with gamma-ray at different levels, were dried and grinded until to obtain a powder. The lipids was extracted with diethyl-ether, and in this solution was measured the total lipid content. The saponins were extracted in a later step, concentrated and precipitated. After the precipitation step the sapogenins were hydrolized and extracted with methyl alcohol. The extracts were centifugated and the solid material was dried and weighted. Finally, the measurement of the oleanolic acid was carried out by high pressure liquid chromatography. One conversion factor permits to relate this acid with the total saponin content. (Author).

Water-soluble derivatives of 25-OCH3-PPD and their anti-proliferative activities.

Science.gov (United States)

Zhou, Wu-Xi; Sun, Yuan-Yuan; Yuan, Wei-Hui; Zhao, Yu-Qing

2017-05-01

(20R)-25-Methoxyl-dammarane-3β,12β,20-triol (25-OCH 3 -PPD, AD-1) is a dammarane-type sapogenin showing anti-tumor potential. In the search for new anti-tumor agents with higher potency than our previously identified compound 25-OCH 3 -PPD, 11 novel sulfamic acid and diacid derivatives that could improve water solubility and contribute to good drug potency and pharmacokinetic profiles were designed and synthesized. Their in vitro anti-tumor activities in MCF-7, A-549, HCT-116, and BGC-823 cell lines and one normal cell line were tested by standard MTT assay. Results showed that compared with compound 25-OCH 3 -PPD, compounds 1, 4, and 5 exhibited higher cytotoxic activity on almost all cell lines, together with lower toxicity in the normal cell. In particular, compound 1 exhibited the best anti-tumor activity in the in vitro assays. The water solubility of 25-OCH 3 -PPD and its derivatives was tested and the results showed that the solubility of 25-OCH 3 -PPD sulfamic acid and diacid derivatives were better than that of 25-OCH 3 -PPD in water, which may provide valuable data for the research and development of new anti-tumor agents. Copyright © 2017 Elsevier Inc. All rights reserved.

[Studies on preparative technology and quantitative determination for extracts of total saponin in roof of Panax japonicus].

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He, Yu-min; Lu, Ke-ming; Yuan, Ding; Zhang, Chang-cheng

2008-11-01

To explore the optimum extraction and purification condition of the total saponins in the root of Panax japonicus (RPJ), and establish its quality control methods. Designed L16 (4(5)) orthogonal test with the extraction rate of total saponins as index, to determine the rational extraction process, and the techniques of water-saturated n-butanol extraction and acetone precipitation were applied to purify the alcohol extract of RPJ. Total saponins were detected by spectrophotometry and its triterpenoidal sapogenin oleanolic acid detected by HPLC. The optimum conditions of total saponins from RPJ was as follows: the material was pulverized, dipped in 60% ethanol aqueous solution as extract solvent at 10 times of volume, and refluxed 3 times for 3 h each time. Extractant of water-saturated n-butanol with extraction times of 3 and precipitant of acetone with precipitation amount of 4-5 times were included in the purification process, which would obtain the quality products. The content of total saponins could reach to 83.48%, and oleanolic acid to 38.30%. The optimized preparative technology is stable, convenient and practical. The extract rate of RPJ was high and steady with this technology, which provided new evidence for industrializing production of the plant and developing new drug.

Synthesis of specifically deuterated pregnanolone and pregnanediol sulphates for metabolic studies in pregant women

International Nuclear Information System (INIS)

Baillie, T.A.; Herz, J.E.; Anderson, R.A.; Sjovall, J.

1975-01-01

A series of specifically deuterated isomers of pregnanolone sulphate and pregnanediol mono- and disulphate have been synthesized for use in a study of the formation and metabolism of C 21 -steroid sulphates in the plasma of pregnant women. Hecogenin, the starting material for the synthesis, was equilibrated with NaO 2 H- 2 H 2 O and the resulting 11,11- 2 H 2 derivative converted directly to its tosylhydrazone. Reduction with sodium borohydride and degradation of the sapogenin side-chain gave 3β-hydroxy-5α-[11,11- 2 H 2 ] pregn-16-en-20-one (isotopic purity 91.6%). Further deuterium atoms were introduced in the 3α and 3α,20β positions by reductions with sodium borodeuteride and lithium aluminum deuteride, respectively. These reactions led to 3β-hydroxy-5α-[3α,11,11- 2 H 3 ] pregnan-20-one (isotopic purity 87.2%) and 5α-[3α,11,11,20β- 2 H 4 ] pregnane-3β,20α-diol (isotopic purity 83.9%). Steroids possessing the less readily accessible 3α-hydroxy-5α-[3β- 2 H] configuration were obtained, without loss of label, by epimerization of the 3β isomers. Pregnanolone sulphates and pregnanediol disulphates were prepared directly from the free alcohols, while the pregnanediol 3-monosulphates were synthesized from the appropriate pregnanediol 20-monoacetate derivatives

Cell wall integrity, genotoxic injury and PCD dynamics in alfalfa saponin-treated white poplar cells highlight a complex link between molecule structure and activity.

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Paparella, Stefania; Tava, Aldo; Avato, Pinarosa; Biazzi, Elisa; Macovei, Anca; Biggiogera, Marco; Carbonera, Daniela; Balestrazzi, Alma

2015-03-01

In the present work, eleven saponins and three sapogenins purified from Medicago sativa were tested for their cytotoxicity against highly proliferating white poplar (Populus alba L.) cell suspension cultures. After preliminary screening, four saponins with different structural features in terms of aglycone moieties and sugar chains (saponin 3, a bidesmoside of hederagenin; saponins 4 and 5, monodesmoside and bidesmoside of medicagenic acid respectively, and saponin 10, a bidesmoside of zanhic acid) and different cytotoxicity were selected and used for further investigation on their structure-activity relationship. Transmission Electron Microscopy (TEM) analyses provided for the first time evidence of the effects exerted by saponins on plant cell wall integrity. Exposure to saponin 3 and saponin 10 resulted into disorganization of the outer wall layer and the effect was even more pronounced in white poplar cells treated with the two medicagenic acid derivatives, saponins 4 and 5. Oxidative burst and nitric oxide accumulation were common hallmarks of the response of white poplar cells to saponins. When DNA damage accumulation and DNA repair profiles were evaluated by Single Cell Gel Electrophoresis, induction of single and double strand breaks followed by effective repair was observed within 24h. The reported data are discussed in view of the current issues dealing with saponin structure-activity relationship. Copyright © 2015 Elsevier Ltd. All rights reserved.

Saponinas esteroidales de la planta Agave brittoniana (Agavaceae con actividad contra el parásito Trichomona vaginalis

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José Orestes Guerra

2008-12-01

the genital tract. To test in vitro the plant against Trichomona vaginalis, the dried and powdered leaves were extracted three times with ethanol-water (7 : 3 by maceration at room temperature. The solvent was removed under reduced pressure and the extract was suspended in distilled water, defatted with n-hexane, and extracted with water-saturated n-butanol. After solvent removal, a portion of the n-butanol extract was hydrolyzed. After extraction with ethyl acetate the hydrolysis products were compared with authentic sapogenins samples using thin layer chromatography (TLC. Most of the sapogenins (yuccagenin and diosgenin were isolated and their structures were confirmed. using nuclear magnetic resonance (NMR experiments. The n-butanol extract was subjected to a separation process through column chromatography to obtain five fractions. After multiple separation processes by reversed phase high performance liquid chromatography (HPLC, the most active one produced one refined fraction that contained two saponins with the same aglycone (diosgenin and one yuccagenin based saponin. Best results of the activity were obtained with the yuccagenin derived glycoside. Rev. Biol. Trop. 56 (4: 16451652. Epub 2008 December 12.

Hemolytic and cytotoxic properties of saponin purified from Holothuria leucospilota sea cucumber.

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Soltani, Mozhgan; Parivar, Kazem; Baharara, Javad; Kerachian, Mohammad Amin; Asili, Javad

2014-10-01

Holothuroids (sea cucumbers) are members of the phylum echinodermata, which produce saponins. Saponins exhibit a wide spectrum of pharmacological and biological activities. In this study, we isolated the crude saponins from the body wall of the dominant Iranian species of sea cucumber, Holothuria leucospilota (H. leucospilota). The purpose of this study was to confirm the presence of saponins in the Persian Gulf H. leucospilota and study the hemolytic and cytotoxic activities of these compounds. The body wall of sea cucumber was dried and powdered and the crude saponins were isolated using various solvents. The crude saponins were further purified by column chromatography using HP-20 resin. The foam test, Thin Layer Chromatography (TLC), hemolytic assay, and Fourier Transform Infrared Spectroscopy (FTIR) confirmed the presence of saponins. Cytotoxicity was analyzed using a 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay on A549 cells, a human lung cancer cell line. The foam test, hemolytic assay, and TLC supported the presence of saponin compounds in the 80% ethanol fraction of H. leucospilota. The infrared (IR) spectrum of the extract showed hydroxyl (-OH), alkyl (C-H), ether (C-O) and ester (-C=O) absorption characteristic of teriterpenoid saponins. The C-O-C absorption indicated glycoside linkages to the sapogenins. The crude saponin extracted from sea cucumber was cytotoxic to A549 cells. The 80% ethanol fraction of saponin isolated from H. leucospilota exhibited hemolytic activity and offers promise as an anti-cancer candidate.

Medicago truncatula CYP716A12 Is a Multifunctional Oxidase Involved in the Biosynthesis of Hemolytic Saponins[W

Science.gov (United States)

Carelli, Maria; Biazzi, Elisa; Panara, Francesco; Tava, Aldo; Scaramelli, Laura; Porceddu, Andrea; Graham, Neil; Odoardi, Miriam; Piano, Efisio; Arcioni, Sergio; May, Sean; Scotti, Carla; Calderini, Ornella

2011-01-01

Saponins, a group of glycosidic compounds present in several plant species, have aglycone moieties that are formed using triterpenoid or steroidal skeletons. In spite of their importance as antimicrobial compounds and their possible benefits for human health, knowledge of the genetic control of saponin biosynthesis is still poorly understood. In the Medicago genus, the hemolytic activity of saponins is related to the nature of their aglycone moieties. We have identified a cytochrome P450 gene (CYP716A12) involved in saponin synthesis in Medicago truncatula using a combined genetic and biochemical approach. Genetic loss-of-function analysis and complementation studies showed that CYP716A12 is responsible for an early step in the saponin biosynthetic pathway. Mutants in CYP716A12 were unable to produce hemolytic saponins and only synthetized soyasaponins, and were thus named lacking hemolytic activity (lha). In vitro enzymatic activity assays indicate that CYP716A12 catalyzes the oxidation of β-amyrin and erythrodiol at the C-28 position, yielding oleanolic acid. Transcriptome changes in the lha mutant showed a modulation in the main steps of triterpenic saponin biosynthetic pathway: squalene cyclization, β-amyrin oxidation, and glycosylation. The analysis of CYP716A12 expression in planta is reported together with the sapogenin content in different tissues and stages. This article provides evidence for CYP716A12 being a key gene in hemolytic saponin biosynthesis. PMID:21821776

Rapid Screening for α-Glucosidase Inhibitors from Gymnema sylvestre by Affinity Ultrafiltration–HPLC-MS

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Mingquan Guo

2017-04-01

Full Text Available Gymnema sylvestre R. Br. (Asclepiadaceae has been known to posses potential anti-diabetic activity, and the gymnemic acids were reported as the main bioactive components in this plant species. However, the specific components responsible for the hypoglycemic effect still remain unknown. In the present study, the in vitro study revealed that the extract of G. sylvestre exhibited significant inhibitory activity against α-glucosidase with IC50 at 68.70 ± 1.22 μg/mL compared to acarbose (positive control at 59.03 ± 2.30 μg/mL, which further indicated the potential anti-diabetic activity. To this end, a method based on affinity ultrafiltration coupled with liquid chromatography mass spectrometry (UF-HPLC-MS was established to rapidly screen and identify the α-glucosidase inhibitors from G. sylvestre. In this way, 9 compounds with higher enrichment factors (EFs were identified according to their MS/MS spectra. Finally, the structure-activity relationships revealed that glycosylation could decrease the potential antisweet activity of sapogenins, and other components except gymnemic acids in G. sylvestre could also be good α-glucosidase inhibitors due to their synergistic effects. Taken together, the proposed method combing α-glucosidase and UF-HPLC-MS presents high efficiency for rapidly screening and identifying potential inhibitors of α-glucosidase from complex natural products, and could be further explored as a valuable high-throughput screening (HTS platform in the early anti-diabetic drug discovery stage.

Medicago truncatula CYP716A12 is a multifunctional oxidase involved in the biosynthesis of hemolytic saponins.

Science.gov (United States)

Carelli, Maria; Biazzi, Elisa; Panara, Francesco; Tava, Aldo; Scaramelli, Laura; Porceddu, Andrea; Graham, Neil; Odoardi, Miriam; Piano, Efisio; Arcioni, Sergio; May, Sean; Scotti, Carla; Calderini, Ornella

2011-08-01

Saponins, a group of glycosidic compounds present in several plant species, have aglycone moieties that are formed using triterpenoid or steroidal skeletons. In spite of their importance as antimicrobial compounds and their possible benefits for human health, knowledge of the genetic control of saponin biosynthesis is still poorly understood. In the Medicago genus, the hemolytic activity of saponins is related to the nature of their aglycone moieties. We have identified a cytochrome P450 gene (CYP716A12) involved in saponin synthesis in Medicago truncatula using a combined genetic and biochemical approach. Genetic loss-of-function analysis and complementation studies showed that CYP716A12 is responsible for an early step in the saponin biosynthetic pathway. Mutants in CYP716A12 were unable to produce hemolytic saponins and only synthetized soyasaponins, and were thus named lacking hemolytic activity (lha). In vitro enzymatic activity assays indicate that CYP716A12 catalyzes the oxidation of β-amyrin and erythrodiol at the C-28 position, yielding oleanolic acid. Transcriptome changes in the lha mutant showed a modulation in the main steps of triterpenic saponin biosynthetic pathway: squalene cyclization, β-amyrin oxidation, and glycosylation. The analysis of CYP716A12 expression in planta is reported together with the sapogenin content in different tissues and stages. This article provides evidence for CYP716A12 being a key gene in hemolytic saponin biosynthesis.

Diosgenin induces apoptosis in IGF-1-stimulated human thyrocytes through two caspase-dependent pathways

International Nuclear Information System (INIS)

Mu, Shumin; Tian, Xingsong; Ruan, Yongwei; Liu, Yuantao; Bian, Dezhi; Ma, Chunyan; Yu, Chunxiao; Feng, Mei; Wang, Furong; Gao, Ling; Zhao, Jia-jun

2012-01-01

Highlights: ► Diosgenin induces apoptosis in IGF-1-treated thyrocytes through two caspase pathways. ► Diosgenin inhibits FLIP and activates caspase-8 in FAS related-pathway. ► Diosgenin increases ROS, regulates the ratio of Bax/Bcl-2 in mitochondrial pathway. -- Abstract: Insulin-like growth factor-1 (IGF-1) is a growth factor of the thyroid that has been shown in our previous study to possess proliferative and antiapoptotic effects in FRTL-5 cell lines through the upregulation of cyclin D and Fas-associated death domain-like interleukin-1-converting enzyme (FLICE)-inhibitory protein (FLIP). Diosgenin, a natural steroid sapogenin from plants, has been shown to induce apoptosis in many cell lines, with the exception of thyroid cells. In this report, we investigated the apoptotic effect and mechanism of diosgenin in IGF-1-stimulated primary human thyrocytes. Primary human thyrocytes were preincubated with or without IGF-1 for 24 h and subsequently exposed to varying concentrations of diosgenin for different times. We found that diosgenin induced apoptosis in human thyrocytes pretreated with IGF-1 in a dose-dependent manner through the activation of caspase cascades. Moreover, diosgenin inhibited FLIP and activated caspase-8 in the FAS-related apoptotic pathway. Diosgenin increased the production of ROS, regulated the balance of Bax and Bcl-2 and cleaved caspase-9 in the mitochondrial apoptotic pathway. These results indicate that diosgenin induces apoptosis in IGF-1-stimulated primary human thyrocytes through two caspase-dependent pathways.

Hemolytic and Cytotoxic Properties of Saponin Purified from Holothuria leucospilota Sea Cucumber

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Mozhgan Soltani

2014-10-01

Full Text Available Background: Holothuroids (sea cucumbers are members of the phylum echinodermata, which produce saponins. Saponins exhibit a wide spectrum of pharmacological and biological activities. In this study, we isolated the crude saponins from the body wall of the dominant Iranian species of sea cucumber, Holothuria leucospilota (H. leucospilota. The purpose of this study was to confirm the presence of saponins in the Persian Gulf H. leucospilota and study the hemolytic and cytotoxic activities of these compounds. Methods: The body wall of sea cucumber was dried and powdered and the crude saponins were isolated using various solvents. The crude saponins were further purified by column chromatography using HP-20 resin. The foam test, Thin Layer Chromatography (TLC, hemolytic assay, and Fourier Transform Infrared Spectroscopy (FTIR confirmed the presence of saponins. Cytotoxicity was analyzed using a 3-(4, 5-dimethylthiazol-2-yl-2, 5-diphenyltetrazolium bromide (MTT assay on A549 cells, a human lung cancer cell line. Results: The foam test, hemolytic assay, and TLC supported the presence of saponin compounds in the 80% ethanol fraction of H. leucospilota. The infrared (IR spectrum of the extract showed hydroxyl (-OH, alkyl (C-H, ether (C-O and ester (–C=O absorption characteristic of teriterpenoid saponins. The C-O-C absorption indicated glycoside linkages to the sapogenins. The crude saponin extracted from sea cucumber was cytotoxic to A549 cells. Conclusion: The 80% ethanol fraction of saponin isolated from H. leucospilota exhibited hemolytic activity and offers promise as an anti-cancer candidate.

Diosgenin induces apoptosis in IGF-1-stimulated human thyrocytes through two caspase-dependent pathways

Energy Technology Data Exchange (ETDEWEB)

Mu, Shumin [Provincial Hospital Affiliated to Shandong University, Jinan 250021 (China); Hospital Affiliated to Shandong Traditional Chinese Medicine University, Jinan 250011 (China); Institute of Endocrinology, Shandong Academy of Clinical Medicine, Jinan 250021 (China); Tian, Xingsong; Ruan, Yongwei [Provincial Hospital Affiliated to Shandong University, Jinan 250021 (China); Liu, Yuantao [The Second Hospital of Shandong University, Jinan 250033 (China); Bian, Dezhi [Provincial Hospital Affiliated to Shandong University, Jinan 250021 (China); Jining Medical College, Jining 272013 (China); Ma, Chunyan [Provincial Hospital Affiliated to Shandong University, Jinan 250021 (China); Yu, Chunxiao [Provincial Hospital Affiliated to Shandong University, Jinan 250021 (China); Institute of Endocrinology, Shandong Academy of Clinical Medicine, Jinan 250021 (China); Feng, Mei [Provincial Hospital Affiliated to Shandong University, Jinan 250021 (China); Wang, Furong [Shandong University of Traditional Chinese Medicine, Jinan 250011 (China); Gao, Ling [Provincial Hospital Affiliated to Shandong University, Jinan 250021 (China); Institute of Endocrinology, Shandong Academy of Clinical Medicine, Jinan 250021 (China); Zhao, Jia-jun, E-mail: jjzhao@medmail.com.cn [Provincial Hospital Affiliated to Shandong University, Jinan 250021 (China); Institute of Endocrinology, Shandong Academy of Clinical Medicine, Jinan 250021 (China)

2012-02-10

Highlights: Black-Right-Pointing-Pointer Diosgenin induces apoptosis in IGF-1-treated thyrocytes through two caspase pathways. Black-Right-Pointing-Pointer Diosgenin inhibits FLIP and activates caspase-8 in FAS related-pathway. Black-Right-Pointing-Pointer Diosgenin increases ROS, regulates the ratio of Bax/Bcl-2 in mitochondrial pathway. -- Abstract: Insulin-like growth factor-1 (IGF-1) is a growth factor of the thyroid that has been shown in our previous study to possess proliferative and antiapoptotic effects in FRTL-5 cell lines through the upregulation of cyclin D and Fas-associated death domain-like interleukin-1-converting enzyme (FLICE)-inhibitory protein (FLIP). Diosgenin, a natural steroid sapogenin from plants, has been shown to induce apoptosis in many cell lines, with the exception of thyroid cells. In this report, we investigated the apoptotic effect and mechanism of diosgenin in IGF-1-stimulated primary human thyrocytes. Primary human thyrocytes were preincubated with or without IGF-1 for 24 h and subsequently exposed to varying concentrations of diosgenin for different times. We found that diosgenin induced apoptosis in human thyrocytes pretreated with IGF-1 in a dose-dependent manner through the activation of caspase cascades. Moreover, diosgenin inhibited FLIP and activated caspase-8 in the FAS-related apoptotic pathway. Diosgenin increased the production of ROS, regulated the balance of Bax and Bcl-2 and cleaved caspase-9 in the mitochondrial apoptotic pathway. These results indicate that diosgenin induces apoptosis in IGF-1-stimulated primary human thyrocytes through two caspase-dependent pathways.

Study and development of three jaboncillo (Sapindus saponaria L.) products as grounding in its industrialisation

International Nuclear Information System (INIS)

Flechas, Henry; Arango, Camilo; Morales, Nelly B; Jimenez, Jhon

2009-01-01

This research included the chemical and phyto chemical studies and the industrial scaling in pilot plant of three products of Sapindus saponaria, commonly known as jaboncillo, Michu. This work is part of an interdisciplinary research project aimed at the exploitation and commercialization of its harvested products: oil, gum, and triterpenoidal sapogenins. These three products were obtained through physical and chemical processes both in laboratory (phase 1) and in the industrial scaling stage (phase 2). During the laboratory stage different methods of extraction and purification were studied and assessed, along with the corresponding methods of chemical analysis for the three origins (Palmira, Valle; Viota, Cundinamarca, and Santa Maria, Huila). Based on laboratory studies, extraction and purification processes were designed in a pilot plant supplied by the QUINACOL Ltda. company, whose business is the production, research and development of products for the phyto pharmaceutical industry. Developed products were characterized by methods of instrumental analysis of nuclear magnetic resonance (1H NMR and 13C), flame ionization detector gas chromatography (GC), infrared spectroscopy (IR) and specific chemical analysis techniques designed for these products. The mature fruit of Santa Maria presented increased performance of oil and rubber; as well as the green fruit of Viota, in a substance called Ss1, possibly hederagenina. Of the products obtained in the pilot phase, oil presented a high percentage of unsaturated fatty acids and rubber showed a high thickening capacity in aqueous solutions. Oil and rubber could be marketed from this species' fruit, because its composition and characteristics, as well as its performance, meets the main parameters of marketing, for which studies on industrial perfectibility are needed.

25-OCH3-PPD induces the apoptosis of activated t-HSC/Cl-6 cells via c-FLIP-mediated NF-κB activation.

Science.gov (United States)

Wu, Yan-ling; Wan, Ying; Jin, Xue-Jun; OuYang, Bing-Qing; Bai, Ting; Zhao, Yu-Qing; Nan, Ji-Xing

2011-11-15

25-OCH(3)-PPD is a dammarane-type triterpene sapogenin isolated from the roots, leaves and seeds of Panax notoginseng, which has shown anti-tumor effects in several human cancer lines. In this study, we evaluated the effects of 25-OCH(3)-PPD on apoptosis of activated t-HSC/Cl-6 cells induced by tumor necrosis factor-α (TNF-α). The inhibitory effects of eleven compounds isolated from Panax ginseng and P. notoginseng were detected in activated t-HSC/Cl-6 cells. 25-OCH(3)-PPD produced a significant inhibitory effect on activated t-HSC/Cl-6 cells. However, 25-OCH(3)-PPD showed almost no effect on the cell viability of Chang liver cells, a type of normal human hepatic cell line. Therefore, we aimed to determine the anti-fibrotic potential of 25-OCH(3)-PPD and to characterize the signal transduction pathways involved in activated HSCs. 25-OCH(3)-PPD decreased the fibrosis markers, including α-smooth muscle actin (α-SMA), transforming growth factor β-1 (TGF-β1) and tissue inhibitors of metalloproteinases-1 (TIMP-1). 25-OCH(3)-PPD elevated the level of cellular GSH in activated HSCs, which demonstrated that 25-OCH(3)-PPD might inhibit HSC activation by its antioxidant capacity. Further analyses revealed that 25-OCH(3)-PPD increased the levels of cleaved caspase-3, decreased the ratio of Bcl-2/Bax and the expression of survivin via c-FLIP-mediated NF-κB activation and shed light on the regulation of apoptosis. Therefore, 25-OCH(3)-PPD may prove to be an excellent candidate agent for the therapy of hepatic fibrosis. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

Spectrum-Effect Relationships between Fingerprints of Caulophyllum robustum Maxim and Inhabited Pro-Inflammation Cytokine Effects.

Science.gov (United States)

Lü, Shaowa; Dong, Shuyu; Xu, Dan; Duan, Jixin; Li, Guoyu; Guo, Yuyan; Kuang, Haixue; Wang, Qiuhong

2017-10-26

Caulophyllum robustum Maxim (CRM) is a Chinese folk medicine with significant effect on treatment of rheumatoid arthritis (RA). This study was designed to explore the spectrum-effect relationships between high-performance liquid chromatography (HPLC) fingerprints and the anti-inflammatory effects of CRM. Seventeen common peaks were detected by fingerprint similarity evaluation software. Among them, 15 peaks were identified by Liquid Chromatography-Mass Spectrometry (LC-MS). Pharmacodynamics experiments were conducted in collagen-induced arthritis (CIA) mice to obtain the anti-inflammatory effects of different batches of CRM with four pro-inflammation cytokines (TNF-α, IL-β, IL-6, and IL-17) as indicators. These cytokines were suppressed at different levels according to the different batches of CRM treatment. The spectrum-effect relationships between chemical fingerprints and the pro-inflammation effects of CRM were established by multiple linear regression (MLR) and gray relational analysis (GRA). The spectrum-effect relationships revealed that the alkaloids ( N -methylcytisine, magnoflorine), saponins (leiyemudanoside C, leiyemudanoside D, leiyemudanoside G, leiyemudanoside B, cauloside H, leonticin D, cauloside G, cauloside D, cauloside B, cauloside C, and cauloside A), sapogenins (oleanolic acid), β-sitosterols, and unknown compounds (X3, X17) together showed anti-inflammatory efficacy. The results also showed that the correlation between saponins and inflammatory factors was significantly closer than that of alkaloids, and saponins linked with less sugar may have higher inhibition effect on pro-inflammatory cytokines in CIA mice. This work provided a general model of the combination of HPLC and anti-inflammatory effects to study the spectrum-effect relationships of CRM, which can be used to discover the active substance and to control the quality of this treatment.

Crystallization-induced dynamic resolution R-epimer from 25-OCH3-PPD epimeric mixture.

Science.gov (United States)

Zhang, Sainan; Tang, Yun; Cao, Jiaqing; Zhao, Chen; Zhao, Yuqing

2015-11-15

25-OCH3-PPD is a promising antitumor dammarane sapogenin isolated from the total saponin-hydrolyzed extract of Panax ginseng berry and Panax notoginseng leaves. 20(R)-25-OCH3-PPD was more potent as an anti-cancer agent than 20(S)-25-OCH3-PPD and epimeric mixture of 25-OCH3-PPD. This paper describes the rapid separation process of the R-epimer of 25-OCH3-PPD from its epimeric mixture by crystallization-induced dynamic resolution (CIDR). The optimized CIDR process was based on single factor analysis and nine well-planned orthogonal design experiments (OA9 matrix). A rapid and sensitive reverse phase high-performance liquid chromatographic (HPLC) method with evaporative light-scattering detector (ELSD) was developed and validated for the quantitation of 25-OCH3-PPD epimeric mixture and crystalline product. Separation and quantitation were achieved with a silica column using a mobile phase consisting of methanol and water (87:13, v/v) at a flow rate of 1.0mL/min. The ELSD detection was performed at 50°C and 3L/min. Under conditions involving 3mL of 95% ethanol, 8% HCl, and a hermetically sealed environment for 72h, the maximum production of 25(R)-OCH3-PPD was achieved with a chemical purity of 97% and a total yield of 87% through the CIDR process. The 25(R)-OCH3-PPD was nearly completely separated from the 220mg 25-OCH3-PPD epimeric mixture. Overall, a simple and steady small-batch purification process for the large-scale production of 25(R)-OCH3-PPD from 25-OCH3-PPD epimeric mixture was developed. Copyright © 2015 Elsevier B.V. All rights reserved.

Improving the antiprotozoal effect of saponins in the rumen by combination with glycosidase inhibiting iminosugars or by modification of their chemical structure

Science.gov (United States)

de la Fuente, Gabriel; Nash, Robert J.; Braganca, Radek; Duval, Stephane; Bouillon, Marc E.; Lahmann, Martina; Newbold, C. Jamie

2017-01-01

The antiprotozoal effect of saponins is transitory, as when saponins are deglycosylated to sapogenins by rumen microorganisms they become inactive. We hypothesised that the combination of saponins with glycosidase-inhibiting iminosugars might potentially increase the effectiveness of saponins over time by preventing their deglycosylation in the rumen. Alternatively, modifying the structure of the saponins by substituting the sugar moiety with other small polar residues might maintain their activity as the sugar substitute would not be enzymatically cleaved. The aim of this in vitro study was to evaluate the acute antiprotozoal effect and the stability of this effect over a 24 h incubation period using ivy saponins, a stevia extract rich in iminosugars, ivy saponins with stevia extract, and a chemically modified ivy saponin, hederagenin bis-succinate (HBS). The effects on fermentation parameters and rumen bacterial communities were also studied. Ivy saponins with stevia and HBS had a greater antiprotozoal effect than ivy saponins, and this effect was maintained after 24 h of incubation (Pstevia extracts was more effective in shifting the fermentation pattern towards higher propionate (+39%) and lower butyrate (-32%) and lower ammonia concentration (-64%) than the extracts incubated separately. HBS caused a decrease in butyrate (-45%) and an increase in propionate (+43%) molar proportions. However, the decrease in ammonia concentration (-42%) observed in the presence of HBS was less than that caused by ivy saponins, either alone or with stevia. Whereas HBS and stevia impacted on bacterial population in terms of community structure, only HBS had an effect in terms of biodiversity (Pstevia and the modified saponin HBS had a strong antiprotozoal effect, although they differed in their effects on fermentation parameters and bacteria communities. Ivy saponins combined with an iminosugar-rich stevia extract and/or HBS should be evaluated to determine their antiprotozoal

Simultaneous determination of notoginsenoside R₁, ginsenoside Rg₁, ginsenoside Re and 20(S) protopanaxatriol in beagle dog plasma by ultra high performance liquid mass spectrometry after oral administration of a Panax notoginseng saponin preparation.

Science.gov (United States)

Wu, Huichao; Liu, Huimin; Bai, Jie; Lu, Yang; Du, Shouying

2015-01-01

20(S) protopanaxatriol is the main metabolite of notoginsenoside R1, ginsenoside Rg1, ginsenoside Re in Panax notoginseng and has significant activities. A ultra high performance liquid mass spectrometry method has been developed and validated for the simultaneous determination of notoginsenoside R₁ (R1), ginsenoside Rg₁ (Rg₁), ginsenoside Re (Re) and 20(S) protopanaxatriol (PPT) in beagle dog plasma after oral administration of a Panax notoginseng saponin preparation. After the addition of the internal standard (digoxin), plasma samples were subjected to liquid-liquid extraction with acetone and methanol and separated on a 100 × 2.1 mm ACQUITY 1.7 μm C₁₈ column (Waters, USA), with acetonitrile and water as the mobile phase, within a runtime of 7.0 min. The analytes were detected without interference in Selected Reaction Monitoring mode with a change in the electrospray ionization from positive to negative. The detection limits were 0.01 to 0.04 mg/L and the calibration curves of the peak areas for the four ingredients were linear over four orders of magnitude with a correlation coefficient greater than 0.9957. The intra-day and inter-day precision values (relative standard deviation, RSD, %) were within 10.25% and 13.51%, respectively, and the accuracy (relative error, RE, %) was less than 7.81%. The validated method was successfully applied to a comparative pharmacokinetic study of four saponins in beagle dogs after oral administration of a Panax Notoginseng Saponins preparation. The pharmacokinetic parameters were calculated with DAS 3.20. The Tmax and Cmax values indicate a dose-dose relationship between the saponins (R1, Rg1, and Re) and their sapogenin (PPT). Copyright © 2014 Elsevier B.V. All rights reserved.

Balanites Aegyptiaca (L.: A Multipurpose Fruit Tree in Savanna Zone Of Western Sudan

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Kamal Eldin Mohammed Fadl

2015-02-01

Full Text Available Underutilized fruit trees play a vital role in food security and economy of the rural population in a number of African countries. Beside the significant important of the underutilized fruit trees in food security and livelihood of the local population many applications can be summarized such as using of leaves for fodder, branches for fencing materials, fire wood and charcoal making, timber for furniture and constructing huts, controlling soil erosion and competing desert encroachments . In spite of their great potential little attention has been given to this species. Balanites aegyptiaca “soap berry tree; thorn tree, desert date” is an important multipurpose trees species in dry land Africa. The tree is a potential source of medicines, pesticides, edible oil, animal feed, nuts, soap, and fuel wood. The edible fruits are rich in saturated fatty acids which are used as cooking oil. The fruit also contains Stereoids (Saponins, Sapogenins, and Disogenins which are used as row materials for industrial production of contraceptive pills and other sexual hormones. The excessive uses of the tree for fruit production and for other uses combined with scarcity of natural regeneration lead to drastic depletion of this species. The desert date tree is adapted to dry and hot climatic environment which are characterized by increasing of land and water resources. However, little information is available about propagation and domestication of this valuable tree species; therefore, studies are needed for sustainable use of underutilized fruit trees in general and for Balanites aegyptiaca in particular. This article aims at highlighting and summarizing information on different aspect of B. aegyptiaca to stimulate the scientist interest in this valuable tree species which is of economical importance for rural inhabitants of western Sudan and other African countries.DOI: http://dx.doi.org/10.3126/ije.v4i1.12188International Journal of Environment Volume-4

Improving the antiprotozoal effect of saponins in the rumen by combination with glycosidase inhibiting iminosugars or by modification of their chemical structure.

Directory of Open Access Journals (Sweden)

Eva Ramos-Morales

Full Text Available The antiprotozoal effect of saponins is transitory, as when saponins are deglycosylated to sapogenins by rumen microorganisms they become inactive. We hypothesised that the combination of saponins with glycosidase-inhibiting iminosugars might potentially increase the effectiveness of saponins over time by preventing their deglycosylation in the rumen. Alternatively, modifying the structure of the saponins by substituting the sugar moiety with other small polar residues might maintain their activity as the sugar substitute would not be enzymatically cleaved. The aim of this in vitro study was to evaluate the acute antiprotozoal effect and the stability of this effect over a 24 h incubation period using ivy saponins, a stevia extract rich in iminosugars, ivy saponins with stevia extract, and a chemically modified ivy saponin, hederagenin bis-succinate (HBS. The effects on fermentation parameters and rumen bacterial communities were also studied. Ivy saponins with stevia and HBS had a greater antiprotozoal effect than ivy saponins, and this effect was maintained after 24 h of incubation (P<0.001. The combination of ivy and stevia extracts was more effective in shifting the fermentation pattern towards higher propionate (+39% and lower butyrate (-32% and lower ammonia concentration (-64% than the extracts incubated separately. HBS caused a decrease in butyrate (-45% and an increase in propionate (+43% molar proportions. However, the decrease in ammonia concentration (-42% observed in the presence of HBS was less than that caused by ivy saponins, either alone or with stevia. Whereas HBS and stevia impacted on bacterial population in terms of community structure, only HBS had an effect in terms of biodiversity (P<0.05. It was concluded that ivy saponins with stevia and the modified saponin HBS had a strong antiprotozoal effect, although they differed in their effects on fermentation parameters and bacteria communities. Ivy saponins combined with an

A strategy for efficient discovery of new natural compounds by integrating orthogonal column chromatography and liquid chromatography/mass spectrometry analysis: Its application in Panax ginseng, Panax quinquefolium and Panax notoginseng to characterize 437 potential new ginsenosides.

Science.gov (United States)

Yang, Wen-zhi; Ye, Min; Qiao, Xue; Liu, Chun-fang; Miao, Wen-juan; Bo, Tao; Tao, Hai-yan; Guo, De-an

2012-08-20

To discover new natural compounds from herbal medicines tends to be more and more difficult. In this paper, a strategy integrating orthogonal column chromatography and liquid chromatography/mass spectrometry (LC/MS) analysis was proposed, and was applied for rapid discovery of new ginsenosides from Panax ginseng (PG), Panax quinquefolium (PQ), and Panax notoginseng (PN). The ginsenosides extracts were fractionated by MCI gel×silica gel orthogonal column chromatography. The fractions were then separated on a C(18) HPLC column, eluted with a three-component mobile phase (CH(3)CN/CH(3)OH/3mM CH(3)COONH(4)H(2)O), and detected by electrospray ionization tandem mass spectrometry. The structures of unknown ginsenosides were elucidated by analyzing negative and positive ion mass spectra, which provided complementary information on the sapogenins and oligosaccharide chains, respectively. A total of 623 comprising 437 potential new ginsenosides were characterized from the ethanol extracts of PG, PQ and PN. New acylations, diversified saccharide chains and C-17 side chains constituted novelty of the newly identified ginsenosides. An interpretation guideline was proposed for structural characterization of unknown ginsenosides by LC/MS. To confirm reliability of this strategy, two targeted unknown trace ginsenosides were obtained in pure form by LC/MS-guided isolation. Based on extensive NMR spectroscopic analysis and other techniques, they were identified as 3-O-[6-O-(E)-butenoyl-β-D-glucopyranosyl(1,2)-β-D-glucopyranosyl]-20(S)-protopanaxadiol-20-O-β-D-glucopyranosyl(1,6)-β-D-glucopyranoside (named ginsenoside IV) and 3-O-β-D-glucopyranosyl(1,2)-β-D-glucopyranosyl-3β,12β,20(S),24(R)-tetra hydroxy-dammar-25-ene-20-O-β-D-glucopyranosyl(1,6)-β-D-glucopyranoside (ginsenoside V), respectively. The fully established structures were consistent with the MS-oriented structural elucidation. This study expanded our understanding on ginsenosides of Panax species, and the