小叶三点金不同萃取相对尖吻蝮蛇毒主要酶类的抑制作用

Inhibitory Effects of Different Extractions of Desmodium microphyllumon Activities of Major Enzymes from Deinagkistrodon acutus Venom

【目的】考察小叶三点金(Desmodium microphyllum)乙醇提取物不同萃取相对尖吻蝮蛇(Deinagkistrodon acutus)毒主要酶类的抑制作用。【方法】采用不同极性溶剂对小叶三点金乙醇提取物进行萃取,检测各萃取相对尖吻蝮蛇毒中蛋白水解酶、磷脂酶A2、透明质酸酶和类凝血酶的抑制作用,再结合气相色谱-质谱法(GC-MS)对抑制作用最强的萃取相进行组分分析。【结果】乙酸乙酯萃取相能够明显抑制尖吻蝮蛇毒中的蛋白水解酶、磷脂酶A2和透明质酸酶和类凝血酶的活性,经GC-MS分析检测出26种成分,其中正十六烷酸、十八碳烯酸、豆甾醇和谷甾醇具有抑制蛇毒活性的作用。【结论】小叶三点金醇提物乙酸乙酯萃取相对尖吻蝮蛇毒中主要酶类具有明显抑制作用,能够作为有效部位用于抗蛇毒功效组分的筛选研究。

[Purposes]To explore inhibitory effects of different extraction portions of Desmodium microphyllum ethanol extracts on activities of major enzymes fromDeinagkistrodon acutus venom.[Methods]The 75%ethanol extracts obtained fromD.microphyllum were futher fractionated by solvents with increasing polarity into five portions including petroleum ether extracts,trichloromethane extracts,ethyl acetate extracts,butyl alcohol extracts,and water extracts.The inhibitory effects of all extracts on proteolytic enzymes,phospholipase A2,hyaluronidase,and thrombin-like enzyme from D.acutus venom were detected,and the extracts with the strongest inhibitory effect was analyzed using gas chromatography-mass spectrometry（GC-MS）.[Findings]The ethyl acetate extracts significantly inhibited the activities of proteolytic enzyme,phospholipase A2,hyaluronidase,and thrombin-like enzyme from D.acutus venom.Based on the analysis of GC-MS,26 components were found in the extracts.Among which,several components were associated with inhibitory activity against snake venom,inculding n-hexadecanoic acid,vaccenic acid,sitosterol,and stigmasterol.[Conclusions]The ethyl acetate portion of ethanol extracts from D.microphyllum significantly inhibited major enzymes from D.acutus venom,and it can be used as an effective source for screening of anti-sanke venom components.