The In Vitro Anti HBV Activity And Immune Regulation Of Gankang Granule

Objective:The aim of this study was to observe the antiviral action in vitro and immune regulation of traditional Chinese medicine Gankang Granule (GKGL) in treating chronic viral hepatitis B induced by hepatitis B virus (HBV) so as to lay the foundation for the optimization process of the subsequent in-depth study of the preparation.Methods:Using HepG2.2.15 cell model, MTT experiment and apoptosis dyeing experiment were made to detect cytotoxicity of GKGL and observe their dose-effect relationship. Through enzyme-linked immunosorbent assay (ELISA), the expression level of HBeAg and HBsAg by HBV was detected. Using fluorescence quantitative PCR technology, quantitative analysis of replication concentration of HBV DNA was determined. The macrophage phagocytosis function and the serum hemolysin level were measured to research the immune adjustment function.Results:The MTT and ELISA test results found that the GKGL could inhibit the growth of HepG2.2.15 cell line and reduce the expression level of HBeAg and HBsAg along with the increase of drug concentrations. On days 3,6,9, the therapy indexes (TI) of GKGL were all larger than 2, which showed that this compound is low-toxic, secure and effective.Time fluorescence quantitative PCR test found that, compared with control group, GKGL had a significant inhibitory effect of HBV DNA (P< 0.05,P<0.01). And with the increase of concentration and action time, its effect presented a clear dosage-effect and time-effect relationship.The animal model of immunological liver injury induced by injecting iv BCG and LPS was built successfully.In these mice model, GKGL played an important role in reducing level of AST and ALT, protecting immunity hepatic injury. The results of mice macrophage phagocytosis function assay showed that the large dosage and medium dosage of GKGL could obviously strengthened the phagocytosis function (P<0.01) so as to encourage the nonspecifc immunity. The mice serum hemolysin tests revealed that the large dosage and medium dosage of GKGL could increase the level of mice serum hemolysin (P<0.05 P<0.01) to promote the pecific humoral immune function.Conclusions:GKGL plays an important role in reducing HBeAg and HBsAg, inhibiting HBV DNA replication and enhancing the mice immune function. This provides a solid basis for our further development on this new compound.