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中国生物工程杂志

China Biotechnology
China Biotechnology  2015, Vol. 35 Issue (11): 29-35    DOI: 10.13523/j.cb.20151105
    
The Inhibition of HBsAg of Natural Compounds and Bortezomib on Primary Hepatocytes of HBV-Tg Mice and the Proteome Analysis
YI Xue-rui1, YUAN You-cheng1, GONG Liang1, ZHANG Xin-rui1,2, LI Na1,3, KONG Xiang-ping1
1. The Liver Center of PLA, No.458 Hospital of PLA, Guangzhou 510600, China;
2. The Department of Infection & Immunity of No.2 Hospital, Tianjin Medical University, Tianjin 300211, China;
3. The Health Deptartment of People's Hospital of Qinhai Province, Xining 810007, China
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Abstract  

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 HBsAg in the supernatant of cells after treated with drugs for 24h by ELISA.Bortezomib with 1μmol/L treated primary liver cells for 24h, 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.RT-PCR 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 RT-PCR,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 HBsAg.Conclusion: Regulation of protein degradation and folding of host cells will be beneficial to the search for new anti-HBV drugs.



Key wordsHBV-Tg mice      Natural compounds      Bortezomib      HSP      Proteome      Proteasome     
Received: 10 August 2015      Published: 24 November 2015
ZTFLH:  Q816  
Cite this article:

YI Xue-rui, YUAN You-cheng, GONG Liang, ZHANG Xin-rui, LI Na, KONG Xiang-ping. The Inhibition of HBsAg of Natural Compounds and Bortezomib on Primary Hepatocytes of HBV-Tg Mice and the Proteome Analysis. China Biotechnology, 2015, 35(11): 29-35.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20151105     OR     https://manu60.magtech.com.cn/biotech/Y2015/V35/I11/29

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