目的研究从狭叶五味子Schisandra lancifolia中分离化合物扁枝杉香豆素(phyllocoumarin)和(-)-表儿茶酸[(-)-epicatechin]的体外抗乙型肝炎病毒(HBV)活性。方法为了筛选和确认扁枝杉香豆素和(-)-表儿茶酸体外抗HBV活性,以HepG2.2.5细胞为体外研究HBV模型,分别用RPMI640完全培养基稀释不同质量浓度的药物作用于细胞,培养3d后收集上清,采用MTT法检测药物对HepG2.2.5细胞的生长影响和ELISA法检测培养上清中HBsAg和HBeAg的水平,评价扁枝杉香豆素和(-)-表儿茶酸对HBsAg和HBeAg的影响。结果扁枝杉香豆素和(-)-表儿茶酸具有一定的体外抗HBV活性,其细胞毒性非常小,CC50均大于200μg/mL。扁枝杉香豆素具有较强的抑制HBsAg和HBeAg分泌作用。阳性对照药物阿德福韦酯也抑制HBVHBsAg和HBeAg分泌,但在相同质量浓度(.6μg/mL)下其抑制作用较扁枝杉香豆素弱。结论狭叶五味子中化合物扁枝杉香豆素和(-)-表儿茶酸具有一定体外抑制HBV HBsAg和HBeAg分泌的作用,从而起到抗HBV作用。

Objective To evaluate anti-HBV activities of phyllocoumarin (Ⅰ) and (-)-epicatechin (Ⅱ) from Schisandra lancifolia in vitro. Methods To screen and confirm the anti-HBV activities of compounds, phyllocoumarin and (-)-picatechin, HepG2.2.15 cells were used as HBV model in vitro. Different concentration of phyllocoumarin and (-)-epicatechin were added to the wells with the growth of HepG2.2.15 cells and the supernatant was collected at the third day. The cytotoxic activity on the growth of HepG2.2.15 cells was detected by MTT. ELISA for HBsAg and HBeAg assays was used to evaluate the effects of phyllocoumarin and (-)-epicatechin on HBsAg and HBeAg secretion. Results Both phyllocoumarin and (-)-epicatechin showed anti-HBV activity and exerted cytotoxicity against HepG2.2.15 cells with CC50200 μg/mL. But phyllocoumarin decreased the levels of HBsAg and HBeAg secretion more effectively than (-)-epicatechin did. Compared with Adefovir Dipivoxil (ADV), phyllocoumarin inhibited the secretion of HBsAg and HBeAg more effectively at the same concentration (1.6 μg/mL). Conclusion Phyllocoumarin and (-)-epicatechin are potential anti-HBV agents ad show lower cytotoxicity.

国家重点基础研究发展计划(973)资助项目(2009CB522306);中国科学院知识创新工程重要方向资助项目(KSCX-YW-R-24)