[Study of the inhibitory effect of cell entry inhibitory ezetimibe on hepatitis B virus in vitro].

Objective: To study the inhibitory effect of ezetimibe in an experimental model of human hepatoma cell line (HepaRG) infected with hepatitis B virus (HBV) positive human serum in vitro. Methods: Mature HepaRG cells were divided into a treatment group (received drugs) and a control group (did not receive drugs). In the ezetimibe prevention experiment, the cells in the treatment group was treated with drugs 2 h before infection and 24 h during infection. In the ezetimibe treatment experiment, the cells in the treatment group were treated with drugs for 6 ~ 10 days continuously after 24 hours of HBV infection. The expression of HBV DNA and intracellular cccDNA in the supernatant was detected by fluorescence quantitative PCR. Hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) content in the cell supernatant were detected by chemiluminescence. Analysis of variance was used to compare the differences between multiple groups. Pairwise comparisons among groups were followed by t- test with normal distribution. P < 0.05 was considered as statistically significant. Results: Ezetimibe prevention experiment showed that compared with control group, the treatment group was added with 20, 60, and 100 μmol/L ezetimibe before and during infection, and the HBV DNA content in the supernatant 2 days before was significantly reduced (P < 0.05) in the treatment group. Compared with the control group, the HBsAg expression level 2 days before was significantly reduced (P < 0.05) with the addition of 60 μmol/L ezetimibe in the treatment group. Compared with the control group, the expression level of intracellular cccDNA was significantly reduced (P < 0.05) after 10 days with the addition of 100μmol/L ezetimibe in the treatment group. Ezetimibe treatment experiment showed that cccDNA content in the cells were significantly lowered with the immediate addition of 60μmol/L ezetimibe 24 hours after infection for 10 days when compared to control group (P < 0.05). Conclusion: Ezetimibe, as a cytosolic inhibitor, has a certain inhibitory effect on hepatitis B virus infection in both prevention and treatment experiment.目的： 在乙型肝炎病毒（HBV）阳性人血清体外感染人肝癌细胞株（HepaRG）的实验模型中进行依折麦布对HBV的抑制作用研究。 方法： 将成熟HepaRG细胞分成加药物的处理组和不加药物的对照组。在依折麦布预防实验中，处理组均是在感染前2 h及感染24 h期间对细胞进行药物处理；在依折麦布治疗实验中，处理组均是在细胞被HBV感染24 h后，立即用药物对其进行连续6~10 d的处理。用荧光定量PCR方法检测上清液中HBV DNA及细胞内cccDNA的表达量，用化学发光法检测细胞上清液中乙型肝炎表面抗原（HBsAg）、乙型肝炎e抗原（HBeAg）的含量。多组间差异比较用方差分析，组间两两对比中服从正态分布的采用t检验，以P < 0.05为差异有统计学意义。 结果： 依折麦布预防实验显示感染前及感染时加20、60、100 μmol/L依折麦布的处理组与对照组相比，处理组前2 d上清液HBV DNA含量明显下降（P < 0.05）；60 μmol/L依折麦布处理组与对照组相比，前2 d上清液中HBsAg表达水平明显下降（P < 0.05）；100 μmol/L依折麦布处理组与对照组相比，10 d后细胞内cccDNA表达水平明显降低（P < 0.05）。依折麦布治疗实验显示细胞被感染24 h后，立即添加60 μmol/L依折麦布处理10 d，细胞中cccDNA含量明显低于对照组（P < 0.05）。 结论： 无论是在预防实验还是治疗实验中，入胞抑制剂依折麦布对HBV的感染均存在一定的抑制作用。.