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《China Biotechnology》 2015-11
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The Inhibition of HBs Ag of Natural Compounds and Bortezomib on Primary Hepatocytes of HBV-Tg Mice and the Proteome Analysis

YI Xue-rui;YUAN You-cheng;GONG Liang;ZHANG Xin-rui;LI Na;KONG Xiang-ping;The Liver Center of PLA,No.458 Hospital of PLA;The Department of Infection &Immunity of No.2 Hospital,Tianjin Medical University;The Health Deptartment of People’s Hospital of Qinhai Province;  
Objective:To compare the inhibitory effects of bortezomib and several natural compounds on HBV in vitro and to explore the molecular mechanism of bortezomib inhibiting HBV in the primary hepatocytes of HBV-Tg mice.Method:Observing toxicity of drugs with different concentrations on HBV-Tg mice primary liver cells and determine the maximum non-toxic concentration through MTT assay.Cultivated and detected the changes of HBs Ag in the supernatant of cells after treated with drugs for 24 h by ELISA.Bortezomib with 1μmol/L treated primary liver cells for 24 h,the total proteins were extracted from the treated cells,and after the proteins were separated by the dimensional gel electrophoresis the differentially expressed protein spots were found.RTPCR and West blot were used to verify the selected differentially expressed proteins.Results:Among the 8 kinds of natural compounds,pristimerin has the strongest inhibition of HBV,with IC50 being 0.43μmol/L.The IC50 of EGCG being 24.9μmol/L was with good safety.21 differential spots were found through analyzing the proteome on primary hepatic protein treated with bortezomib,and the HSP60 isomers(gi/26353954) was determined in the meantime.The gene expression of HSP60 and 90 was down-regulated while HSP70 was up-regulated with RTPCR,but HSP60 changed little in the protein level with West blot after treated bortezomib on HBV-Tg primary hepatocytes.On the other hand,the inhibition of HSP90 by EGCG was consistent with that of HBs Ag.Conclusion:Regulation of protein degradation and folding of host cells will be beneficial to the search for new anti-HBV drugs.
【Fund】: 国家“艾滋病和病毒性肝炎等重大传染病防治”科技重大专项(2012ZX10004503-010);; 广东省科技计划(2013B060300015)资助项目
【CateGory Index】: R965
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