基于GC-MS对牛奶菜治疗蛇伤作用机制的初步研究
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摘要
【目的】初步考察牛奶菜(Marsdenia sinensis)治疗蛇伤作用机制。【方法】采用乙醇回流提取法和水回流提取法制备牛奶菜提取物,再采用石油醚、三氯甲烷、乙酸乙酯、正丁醇对水提物依次萃取,对各萃取物进行气相色谱-质谱联用法(GC-MS)分析。【结果】从牛奶菜中共检测出77种组分,其中含有大量具有抑制蛇毒活性的组分,包括十四烷酸、十六烷酸、十六烷酸衍生物、十八烷酸衍生物、十八烯酸及它的衍生物、2,4-二叔丁基苯酚、谷甾醇、豆甾醇等。【结论】牛奶菜主要通过抑制蛇毒氧化损伤以及抑制蛇毒神经毒素发挥治疗功效。
[Purposes]To preliminarily investigate the therapeutic mechanism of Marsdenia sinensis on snake envenomation.[Methods]M.sinensis was extracted by the methods of alcohol and water reflux extraction.Then the water extract was futher fractionated by solvents with increasing polarity in order to obtain the petroleum ether,trichloromethane,ethyl acetate,and butyl alcohol fractions.The five kinds of extracts of M.sinensis were analyzed by gas chromatography-mass spectrometry(GC-MS).[Findings]Based on the analysis from GC-MS,77 components were found in this plant,and several components associated with inhibition activity against snake venom were detected in the extract of M.sinensis including tetradecanoic acid,hexadecanoic acid,hexadecanoic acidderivatives,octadecanoic acid derivatives,octadecenoic acid and its derivatives,2,4-bis tert-butyl phenol,sitosterol,and stigmasterol.[Conclusions]It seems reasonable to suppose that M.sinensis could funtion in preventing the oxidative damage induced by snakebites and inhibiting snake venom neurotoxicity,resulting in therapeutic properties of snake envenomation.
[Purposes]To preliminarily investigate the therapeutic mechanism of Marsdenia sinensis on snake envenomation.[Methods]M.sinensis was extracted by the methods of alcohol and water reflux extraction.Then the water extract was futher fractionated by solvents with increasing polarity in order to obtain the petroleum ether,trichloromethane,ethyl acetate,and butyl alcohol fractions.The five kinds of extracts of M.sinensis were analyzed by gas chromatography-mass spectrometry(GC-MS).[Findings]Based on the analysis from GC-MS,77 components were found in this plant,and several components associated with inhibition activity against snake venom were detected in the extract of M.sinensis including tetradecanoic acid,hexadecanoic acid,hexadecanoic acidderivatives,octadecanoic acid derivatives,octadecenoic acid and its derivatives,2,4-bis tert-butyl phenol,sitosterol,and stigmasterol.[Conclusions]It seems reasonable to suppose that M.sinensis could funtion in preventing the oxidative damage induced by snakebites and inhibiting snake venom neurotoxicity,resulting in therapeutic properties of snake envenomation.
引文
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[12]LEE C Y,CHEN Y M,JOUBERT F J.Protection by atropine against synergistic lethal effects of the angusticepstype toxin F7from eastern green mamba venom and toxin I from black mamba venom[J].Toxicon Official Journal of the International Society on Toxinology,1982,20(3):665-667.
[13]DELATORRE P,ROCHA B A,SANTI-GADELHA T,et al.Crystal structure of Bn IV in complex with myristic acid:a Lys49 myotoxic phospholipase A2from Bothrops neuwiedi venom[J].Biochimie,2011,93(3):513-518.
[14]VICKERS C E,POSSELL M,COJOCARIU C I,et al.Isoprene synthesis protects transgenic tobacco plants from oxidative stress[J].Plant Cell and Environment,2009,32(5):520-531.
[15]APARNA V,DILEEP K V,MANDAL P K,et al.Antiinflammatory property of n-hexadecanoic acid:structural evidence and kinetic assessment[J].Chemical Biology&Drug Design,2012,80(3):434-439.
[16]BELGHIT S,DRICHE E H,BIJANI C,et al.Activity of2,4-Di-tert-butylphenol produced by a strain of Streptomyces mutabilis isolated from a Saharan soil against Candida albicans and other pathogenic fungi[J].Journal De Mycologie Medicale,2016,26(2):160-169.
[17]GOMES A,SAHA A,CHATTERJEE I,et al.Viper and cobra venom neutralization by beta-sitosterol and stigmasterol isolated from the root extract of Pluchea indica Less.(Asteraceae)[J].Phytomedicine International Journal of Phytotherapy&Phytopharmacology,2007,14(9):637-643.
[2]GUTIéRREZ J M,LEóN G,BURNOUF T.Antivenoms for the treatment of snakebite envenomings:the road ahead[J].Biologicals Journal of the International Association of Biological Standardization,2011,39(3):129-142.
[3]SOARES A M,TICLI F K,MARCUSSI S,et al.Medicinal plants with inhibitory properties against snake venoms[J].Current Medicinal Chemistry,2005,12(22):2625-2641.
[4]GOMES A,DAS R,SARKHEL S,et al.Herbs and herbal constituents active against snake bite[J].Indian Journal of Experimental Biology,2010,48(9):865-878.
[5]顾海峰,梁晋鄂,李春美,等.柿子单宁对几种蛇毒中主要酶的抑制作用及其机理初探[J].中国农业科学,2008,41(3):910-917.GU H F,LIANG J E,LI C M,et al.Study on inhibitory effects of persimmon tannins on activities of major enzymes from several snake venoms and its mechanism[J].Scientia Agricultura Sinica,2008,41(3):910-917.
[6]和七一,熊广均,李博,等.中草药徐长卿(Cynanchum paniculatum)治疗毒蛇咬伤的分子机制研究[J].中国科学:生命科学,2017,47(2):218-229.HE Q Y,XIONG G J,LI B,et al.Study on the molecular mechanism of Chinese herbal medicine Cynanchum paniculatumfor treating snakebites[J].Scientia Sinica Vitae,2017,47(2):218-229.
[7]刘丽仙.对牛奶菜原植物描述的补充和修正[J].湖南中医药大学学报,1985(2):37-38.LIU L X.Supplement and correction of the original plant description of Marsdenia sinensis Hemsl[J].Journal of Traditional Chinese Medicine University of Hunan,1985(2):37-38.
[8]田文艺,兰芳,张昌平,等.毛萝藦注射液对蛇毒中毒动物保护作用的研究[J].湖南中医药大学学报,1985(1):54-55.TIAN W Y,LAN F,ZHANG C P,et al.Study on the protective effect of Marsdenia sinensis Hemsl injection on snake venom poisoning animals[J].Journal of Traditional Chinese Medicine University of Hunan,1985(1):54-55.
[9]杜方麓,田文艺.毛萝藦中各部位对眼镜蛇毒中毒小鼠的保护作用研究[J].湖南中医药大学学报,1994(3):51-52.DU F L,TIAN W Y.Study on the protective effect of different parts of Marsdenia sinensis Hemsl on mice poisoned by cobra Venom[J].Journal of Traditional Chinese Medicine University of Hunan,1994(3):51-52.
[10]MARKLAND F S.Snake venoms and the hemostatic system[J].Toxicon,1998,36(12):1749-1800.
[11]ALCARAZ M J,HOULT J R.Effects of hypolaetin-8-glucoside and related flavonoids on soybean lipoxygenase and snake venom phospholipase A2[J].Archives Internationales de Pharmacodynamie et de Thérapie,1985,278(1):4-12.
[12]LEE C Y,CHEN Y M,JOUBERT F J.Protection by atropine against synergistic lethal effects of the angusticepstype toxin F7from eastern green mamba venom and toxin I from black mamba venom[J].Toxicon Official Journal of the International Society on Toxinology,1982,20(3):665-667.
[13]DELATORRE P,ROCHA B A,SANTI-GADELHA T,et al.Crystal structure of Bn IV in complex with myristic acid:a Lys49 myotoxic phospholipase A2from Bothrops neuwiedi venom[J].Biochimie,2011,93(3):513-518.
[14]VICKERS C E,POSSELL M,COJOCARIU C I,et al.Isoprene synthesis protects transgenic tobacco plants from oxidative stress[J].Plant Cell and Environment,2009,32(5):520-531.
[15]APARNA V,DILEEP K V,MANDAL P K,et al.Antiinflammatory property of n-hexadecanoic acid:structural evidence and kinetic assessment[J].Chemical Biology&Drug Design,2012,80(3):434-439.
[16]BELGHIT S,DRICHE E H,BIJANI C,et al.Activity of2,4-Di-tert-butylphenol produced by a strain of Streptomyces mutabilis isolated from a Saharan soil against Candida albicans and other pathogenic fungi[J].Journal De Mycologie Medicale,2016,26(2):160-169.
[17]GOMES A,SAHA A,CHATTERJEE I,et al.Viper and cobra venom neutralization by beta-sitosterol and stigmasterol isolated from the root extract of Pluchea indica Less.(Asteraceae)[J].Phytomedicine International Journal of Phytotherapy&Phytopharmacology,2007,14(9):637-643.
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