山豆根中皂苷等不同组分的体外抗HBV实验研究

Analysis of the saponins in Tonkinensis and its anti-HBV in vitro

目的揭示山豆根中抗HBV的物质基础,并探索建立肝炎灵新的质控标准。方法采用正丁醇萃取方法提取山豆根药材中的总皂苷成分。并以HepG2.2.15细胞株为模型,实验分为空白组(不接种细胞只加培养液)、细胞对照组(接种细胞但不加药物)、药物干预组(分为不同浓度组)、阳性药物(拉米夫丁)对照组,药物作用9天后,采用ELISA法检测药物作用后细胞培养上清液中HBsAg、HBeAg,采用荧光定量PCR法测定上清中病毒DNA含量,采用MTT法检测细胞毒性。结果山豆根中皂苷类组分和其他各组分在体外只有高浓度时对HepH2.2.15细胞具有毒性抑制作用,其他稀释倍数的浓度均无显著毒性;山豆根中皂苷类组分和其他各组在体外都具有一定的抑制HepG2.2.15细胞分泌HBsAg、HBeAg作用,并呈一定的剂量依赖性,各组分之间无显著差异性,其中山豆根中正丁醇水溶液组分的抑制HepG2.2.15细胞分泌HBsAg、HBeAg的作用较优,而山豆根中皂苷和其他组分并未显示出更好的抑制作用。结论山豆根中皂苷和其他组分并未显示出更好的抑制病毒作用,提示山豆根中尚有其他成分或各种不同成分组合的抗HBV作用更佳。

Objective Through the analysis of saponins in tonkinensis and its experimental study of anti-HBV,to reveal its material basis.Methods The total saponins of tonkinensis Radix was extracted with n-butanol extraction.The contents of saponins was measured with exclusive chromogenic agent for saponins.The model of toxic liver injury in mouse was constructed with intraperitoneal injection of CCl4.The mice were divided into normal control group,model control group,hepatitis spiritual group,saponins high,medium and low-dose group.The mice were killed for the observation of liver function and liver histological changes.Results Saponins were rich in tonkinensis herbs.Compared with model group,Ganyanling group showed the obvious difference in downregulating ALT and AST（P〈0.05）.Different doses of saponin group downregulating ALT,AST was more obvious,and the best effect is in the medium dose group（P〈0.05）.Conclusion Saponins are rich in tonkinensis herbs,but almost non in the injection of tonkinensis.Saponins in tonkinensis shows a good chemical protective effect on CCL4-induced liver injury model,but not better than tonkinensis injection（Ganyanling）,suggesting that there are other ingredients in tonkinensis with better effect for anti-liver injury.