3种抗HBV核苷(酸)类似物体外抗乙肝病毒作用

比较3种抗HBV核苷(酸)类似物(拉米夫定、恩替卡韦、替诺福韦)对HepG 2.2.15细胞的毒性作用和对HBsAg、HBeAg和HBV-DNA表达的抑制效果。用四甲基噻唑蓝(MTT)法检测药物的细胞毒性;酶联免疫(ELISA)法检测HBsAg和HBeAg表达水平;PCR-荧光探针法定量检测HBV-DNA含量。结果:在最高浓度200 mg/L时,拉米夫定组存活率为74.72%,恩替卡韦组存活率仅为36.74%,替诺福韦组存活率为95.22%;拉米夫定和替诺福韦对HBsAg、HBeAg表达无明显抑制,恩替卡韦对HBsAg、HBeAg表达有抑制并与其细胞毒性相一致。在第9天,3种药物对HBV-DNA抑制作用明显,在6.25 mg/L浓度时,抑制率均高于90%;3种药物在相同药物浓度下,恩替卡韦对HepG 2.2.15细胞毒性最大,替诺福韦几乎没有细胞毒性,拉米夫定有一定毒性。

Effect of three anti-HBV nucleoside(acid) analogues on hepatitis B virus in vitro

The comparison on the toxicity of three anti-HBV nucleoside(acid) analogues(lamivudine, entecavir and tenofovir) on HepG 2.2.15 cells and their inhibitory effects on the expression of HBsAg, HBeAg and HBV-DNA is carried out Tetramethylthiazole blue(MTT) assay is used to detect the cytotoxicity of drugs, enzyme-linked immunosorbent assay(ELISA) is adopted to detect the expression level of HBsAg and HBeAg, and quantitative detection of HBV-DNA content by PCR-fluorescence probe is conducted. The results are that at the highest concentration of 200 mg/L, the survival rate of lamivudine group is 74.72%, the survival rate of entecavir group is 36.74%, and the survival rate of tenofovir group is 95.22%. Lamivudine and tenofovir don’t inhibit the expression of HBs Ag and HBe Ag, but entecavir has inhibited the expression of HBsAg and HBeAg, which is consistent with their cytotoxicity. In the 9 th day of PCR experiment, the inhibition of HBV-DNA by three drugs is obvious, and at 6.25 mg/L concentration, the inhibition rate is higher than 90%. Under the same concentration of three drugs, entecavir has the greatest cytotoxicity to HepG 2.2.15, tenofovir had almost no cytotoxicity, lamivudine had certain toxicity.