论文部分内容阅读
目的:筛选红背叶根体外抑制丙型肝炎病毒(HCV)复制的活性部位。方法:以HCV亚基因复制子体外培养体系CBRH7919(Jneo3-5B)为HCV复制模型,选用干扰素α联合利巴韦林为阳性对照药物,观察不同浓度红背叶根总提取物、石油醚部位、乙酸乙酯部位、正丁醇部位的抗HCV作用。采用荧光定量PCR检测HCV亚基因复制子RNA拷贝数,通过Western blot检测HCV功能蛋白NS3的表达,采用CCK-8法观察药物对细胞生长的影响。结果:红背叶根4种提取物中,乙酸乙酯部位对HCVRNA复制的抑制活性最强,对NS3蛋白表达有明显干预作用,对HCVRNA表达表现出剂量依赖的抑制作用(P<0.05)。其比活较初始总提取物提高5.71倍,对HCVRNA半数抑制浓度为14.60mg/L,对CBRH7919(Jneo3-5B)细胞半数细胞毒性浓度为40.30mg/L,治疗指数为2.76。结论:红背叶根乙酸乙酯部位可抑制HCV的复制,为抗HCVRNA表达的活性部位,在抗HCV方面有着潜在的用途。
OBJECTIVE: To screen the active site that inhibits the replication of hepatitis C virus (HCV) in vitro by red dorsal root. Methods: The HCV subgenomic replicon in vitro culture system CBRH7919 (Jneo3-5B) for the HCV replication model, the selection of interferon α and ribavirin as positive control drug, observed the different concentrations of red back root extract, petroleum ether site , Ethyl acetate, n-butanol anti-HCV role. The copy number of HCV subgenomic replicon RNA was detected by real-time PCR. The expression of HCV NS3 protein was detected by Western blot. The effect of drugs on cell growth was observed by CCK-8. Results: Ethyl acetate fraction showed the strongest inhibitory effect on the replication of HCV RNA among the four extracts of red dorsal root. It had a significant effect on the expression of NS3 protein and a dose-dependent inhibition of HCV RNA expression (P <0.05). The specific activity was 5.71 fold higher than that of the original total extract. The half inhibitory concentration of HCVRNA was 14.60 mg / L, the half cytotoxic concentration of CBRH7919 (Jneo3-5B) was 40.30 mg / L and the therapeutic index was 2.76. CONCLUSION: The ethyl acetate fraction of red dorsal root inhibits the replication of HCV, which has potential applications in anti-HCV as an active site against HCV RNA expression.