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目的:考察雷公藤红素对HBV转基因小鼠原代肝细胞的抗HBV作用。方法:改良的两步灌流法分离HBV转基因小鼠原代肝细胞;MTT法检测细胞毒性作用;药物作用24 h后取上清,分别应用ELISA法和荧光定量PCR法测定HBsAg和HBV-DNA。结果:雷公藤红素对原代肝细胞TC50为(6.49±0.8)μmol·L-1;雷公藤红素浓度低于0.8μmol·L-1时,对原代肝细胞没有明显毒性;在0.08～0.8μmol·L-1浓度范围内对HBsAg和HBV-DNA有显著抑制作用(P<0.05);随着雷公藤红素浓度的增加,对HBsAg和HBV-DNA的抑制作用增强。结论:雷公藤红素不仅能够抑制转基因小鼠原代肝细胞HBV-DNA的复制,而且可以有效抑制其HBsAg的表达。
Objective: To investigate the anti-HBV effect of tripterine on primary hepatocytes of HBV transgenic mice. Methods: Primary liver cells of HBV transgenic mice were isolated by modified two-step perfusion method. Cytotoxicity was detected by MTT assay. Supernatants were harvested 24 hours after drug treatment. ELISA and fluorescent quantitative PCR were used to detect HBsAg and HBV-DNA respectively. Results: Tripterine had no significant cytotoxicity on primary hepatocytes when TC50 was (6.49 ± 0.8) μmol·L-1 and tripterine concentration was lower than 0.8 μmol·L-1. ~ 0.8 μmol·L-1 concentration (P <0.05). The inhibitory effect of HBsAg and HBV-DNA was enhanced with the increasing of tripterine concentration. Conclusion: Tripterine not only can inhibit the replication of HBV-DNA in the primary hepatocytes of transgenic mice, but also can effectively inhibit the expression of HBsAg.