叶下珠提取物在体外HBV瞬时转染模型中抑制HBV复制及表达的实验研究

o observe the effects of the extract from Phyllanthus urinaria L. on HBV replication and expression in HBV transient transfection model in vitro. Methods The eukaryotic expression plasmid pHBV1.1, which contains 1.1-fold-overlength genome of HBV, was transfected into the human hepatoma cell line, HepG2, to establish and assess the HBV transient transfection model. The extract from Phyllanthus urinaria L. was prepared in different oncentration and MTT method was used to detect the maximum nontoxic concentration of the drug. The extract from Phyllanthus urinaria L. in the concentration of 0.8 g/ L、0.2 g/ L、0.05 g/ L, respectively, was added into the transfected cells. On the 4th day after drug application, ELISA method was used to detect the concentration of HBsAg in the supernatants; Southern blot was used to detect the level and HBV DNA; Western blot was used to detect the expression of HBcAg in cells. Results After the transfection of plasmid pHBV1.1 into HepG2 cells, the concentration of HBsAg in supernatants was increased obviously compared to the normal cells(P<0.05), and all expected HBV replicative intermediates were verified by Southern blot analysis, which demonstrated that the HBV transient transfection model was established successfully. After the application of drugs with the concentration of 0.8 g/ L、0.2 g/ L, the expression level of HBsAg was obviously decreased in the supernatants, compared to the virus group (P<0.05); Southern blot showed that the level of HBV rc DNA, ds DNA, ss DNA was obviously reduced compared to the virus group (P<0.05); Western blot showed the expression HBcAg in the drug group was obviously inhibited, compared to the virus group (P<0.05). Conclusion The extract from Phyllanthus urinaria L. obviously inhibited HBV replication and expression in HBV transfected cell lines in vitro, and exerted distinctive anti-HBV effects.?????