Research And Development Of Esculetin As An Anti-hepatitis B Virus Drug

Objective:To study the inhibitory effects of esculetin from the Microsorium fortunei on hepatitis B virus?HBV?and duck hepatitis B virus?DHBV?in vitro and in vivo.Methods:?1?Anti-hepatitis B virus experiment in vitro:HepG2.2.15cell line was used as cell model.MTT assay was used to evaluate the cytotoxicity effect of esculetin from microsorium fortunei on HepG2.2.15 cell.The levels of HBsAg,HBeAg and HBV DNA in HepG2.2.15 cell supernatant were detected by ELISA and fluorescence quantification PCR.?2?Anti-hepatitis B virus experiment in vivo:five-day-old Guangxi brown spot ducklings with congenital infection of DHBV determined by quantitative real-time PCR were used to establish a duck hepatitis B animal model in the study.The levels of DHBV DNA,Alanine aminotransferase,Aspartate aminotransferase,DHBsAg and DHBeAg in the duck serum were measured to evaluate the inhibitory effect of positive drugs?lamivudine?and esculetin on DHBV in vivo.At the end of the experiment,the liver tissue of the duck was taken for pathological examination to determine the protective effect of the esculetin the liver of the positively infected duck.Results:?1?Esculetin has anti-HBV effect in vitro:Esculetin was less toxic to HepG2.2.15cells,and its TC₅₀?the 50%cytotoxic concentration?was 273.08 mg/L.When treated the cells with 80 mg/L of esculetin for 9 days,the inhibition rates on HBsAg,HBeAg and HBV DNA were?94.74±2.19?%??93.88±2.36?%??90.95±0.67?%,respectively.And there was a significant difference between the experiments group and the negative control group.(^*P?0.05;^**P?0.01).TI>2,the results show that esculetin has anti-HBV effect in vitro,and it is highly effective and low toxicity.?2?Esculetin has anti-HBV effect in vivo:After treatment,each dose of esculetin significantly reduced the levels of DHBV DNA,ALT,AST,DHBsAg,and DHBeAg in the serum of positively infected ducks.compared with the model control group(^*P?0.05;^**P?0.01).Moreover,high doses of esculetin significantly improve the symptoms of inflammation in the liver.Conclusions:The esculetin isolated from Microsorium fortunei are not only inhibit hepatitis B virus in vitro,but also have better anti-duck hepatitis B virus activity in vivo and protective effect on liver damage caused by DHBV.