Study On The Anti-HBV Effect And Its Mechanism Of Corilagin Extracted From Phyllanthus Urinaria L.

Objective: Phyllanthus urinaria L.(PUL)has the functions of calming the liver and clearing heat,purifying the poison,improving the eyesight and eliminating the product.,and is an effective anti-HBV drug.Chronic hepatitis B virus(HBV)infection is an important global health problem.Therefore,the purpose of this study is to explore the anti-HBV effect of the extract of PUL.,extract and analyze the effective components of PUL.,and explore its effect on cell growth and anti-HBV effect.Methods: Hep G2.2.15 cells were taken as the research object and PUL.as the target drug.Based on the identification of the main components of the PUL.,using intelligent super-computing innovative drug virtual screening technology system,screening the target of the main components of the PUL.under the target from more than 2,000 potential drug targets,establish the network relationship between the target to the target,the target to the disease,designed to extract each part of herbal medicine PUL.by pharmacochemical method,then detect the total phenol content of each part.Chemiluminescence method was used to detect HBs Ag and HBe Ag,and the effective components were analyzed by HPLC.The effective monomer components of PUL.were determined,and further research was carried out as the research object.CCK-8assay,monoclonal cell growth,cell cycle detection and apoptosis detection were used to analyze the effects of monomer drugs on cell growth.Chemiluminescence method was used again to detect the effect of monomeric drugs on the secretion of HBs Ag and HBe Ag in Hep G2.2.15 cells,and the corresponding indicators of oxidative stress were explored to explore the possibility of anti-HBV of monomeric drugs on cells.Finally,Target Scan,RNA-Seq and RT-PCR techniques were used to detect the effect of monomer drugs on ID3 gene,and to explore the mechanism of drug action.Results: A total of 8 components were extracted from PUL.,among them,the content of phenolic substances in ethyl acetate fraction is higher,and Ethyl acetate fraction from the extracts of PUL.has a better anti-HBV effect.The results of HPLC showed that Corilagin was contained in the ethyl acetate fraction,and the corresponding literature reported that Corilagin had anti-HBV effect.Therefore,Corilagin was selected as the effective monomer of PUL.and subsequent studies were carried out.Corilagin inhibited the proliferation of Hep G2.2.15 cells in a concentration-dependent manner(P < 0.05).Corilagin can block the cell cycle of Hep G2.2.15,induce apoptosis of Hep G2.2.15 cells,and inhibit the growth of Hep G2.2.15 cells.Corilagin can inhibit the secretion of HBs Ag and HBe Ag by Hep G2.2.15 cells in a concentrationdependent manner(P < 0.05),and inhibit the oxidative stress of Hep G2.2.15 cells.Corilagin may play an anti-HBV role by inhibiting the oxidative stress of Hep G2.2.15 cells.Through RNA-Seq sequencing,we found that Corilagin could inhibit ID3 gene expression(consistent with the validation results).Target Scan predicted that the binding site of 3’-UTR of ID3 gene was micro RNA-224.Corilagin could inhibit ID3 gene expression through micro RNA-224.According to KEGG pathway enrichment analysis,Corilagin downregulated Samd1/5/8 in TGF-beta signaling pathway to inhibit ID3 gene expression.Conclusion: This study showed that PUL.has anti-HBV effect.Corilagin,as an effective monomer of PUL.has the effect of inhibiting the growth of Hep G2.2.15 cells and anti-HBV virus,which provides a basis for the development of new drugs for the treatment of hepatitis B.