摘要
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 effect and the molecular targets of anti-hepatitis B virus (HBV) by β -L-D4A in vitro. Methods 2.2.15 cells were cultured and treated with various concentrations of β -L-D4A for 6 hours, then the effect of anti-HBV was examined by Southern blot and the replicating core particles from the cells were isolated. The endogenous polymerase reaction and activity gel experiment were performed to monitor the activities of the DNA polymerase and reverse transcriptase. Results The replication of HBV DNA was inhibited in a dose-dependent manner. The endogenous polymerase reaction showed both the two enzymatic activities were irreversibly inactivated in a concentration -dependent manner, with IC50 at 0.51μmol/L and 0.55 μmol/L, respectively. But the activities of DNA polymerase and reverse transcriptase were found to remain active by activity gel with exogenous templates. Conclusion The mechanism of inhibiting HBV replication by β -L-D4A may be in that either the DNA replication priming is blocked or the elongation of DNA chain is terminated irreversibly.
出处
《中华肝脏病杂志》
CAS
CSCD
2003年第5期268-270,共3页
Chinese Journal of Hepatology
基金
国家自然科学基金(39970858)
