Abstract: OBJECTIVE   To investigate the anti-hepatitis B virus (HBV) effect of Compound Huzhang Prescription in vivo and in vitro.METHODS   In vitro, CCK-8 staining method was used to determine the anti-proliferative activity of Compound Huzhang Prescription against HepG2.2.15 cell lines expressing HBV, chemiluminescent microparticle immunoassay method was used to determine hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) levels, and fluorogenic quantitative PCR method was used to determine HBV-DNA level. In vivo, Dot blot assay was used to investigate the content of DHBV-DNA in serum at the 5th, 10th day after administration and the 3th day after drug withdrawal.RESULTS   The results showed that water extract and alcohol extract of Compound Huzhang Prescription had a good inhibitory effect on the growth of HepG2.2.15 cells, with a half inhibitory concentration (IC₅₀) of 51.28 mg/mL and 46.78 mg/mL, respectively. Compound Huzhang Prescription significantly inhibited HBsAg and HBeAg expressions and it showed a significant dose effect relationship. The inhibition of alcohol extract was stronger than that of water extract. The alcohol extract had an inhibitory effect on HBV-DNA replication, but the effect of water extract was not obvious. Compound Huzhang Prescription effectively inhibited DHBV-DNA replication, and significantly inhibited the virus rebound after drug withdrawal in vivo.CONCLUSION   Compound Huzhang Prescription shows a certain anti-hepatitis B virus effect in vivo and in vitro, which lays a foundation for the development and application.