论文部分内容阅读
目的 探索β-L-D4A体外抗乙型肝炎病毒(HBV)作用及其靶点,以明确其抗HBV复制作用的机制。 方法 β-L-D4A干预培养2.2.15细胞后,检测其抗HBV作用;提取细胞内复制核心颗粒,通过内源性聚合酶反应和活性胶实验检测聚合酶和逆转录酶的活性。 结果 β-L-D4A体外明显抑制HBV DNA的复制,并呈现浓度依赖性;内源性聚合酶反应显示β-L-D4A对聚合酶和逆转录酶活性均存在抑制作用,其IC50分别为0.51 μmol/L和0.5 5 μmol/L;外源性核酸为模板的活性胶实验显示聚合酶和逆转录酶活性无变化。 结论 β-L-D4A体外抗HBV作用环节可能在于其代谢物对HBV DNA复制的始动或DNA链延伸过程的不可逆抑制。需行引物结合实验进一步验证。
Objective To explore the anti-hepatitis B virus (HBV) effect of β-L-D4A and its target in order to clarify its anti-HBV replication mechanism. Methods β-L-D4A was used to inoculate 2.2.15 cells and the anti-HBV activity was detected. The intracellular replication of core particles was extracted and the activity of polymerase and reverse transcriptase was detected by endogenous polymerase reaction and activity gel assay. Results β-L-D4A inhibited the replication of HBV DNA in vitro in a concentration-dependent manner. The endogenous polymerase reaction showed that β-L-D4A inhibited the activity of both polymerase and reverse transcriptase, with IC50 of 0.51 μmol / L and 0.5 5 μmol / L. Activity experiments with exogenous nucleic acids as templates showed no change in the activity of polymerase and reverse transcriptase. Conclusion The anti-HBV effect of β-L-D4A in vitro may be due to its irreversible inhibition of HBV DNA replication or DNA strand elongation. Need to be combined with the primer further verified.