Comparative Proteomics Analysis Of Hepatitis B Virus Related Hepatic Fibrosis Tissues

Objective: In this study, we tried to apply the fluorescence differential in-gel electrophoresis (DIGE) technique for the identification of HBV related fibrosis specific protein markers and for the exploration of new molecular markers for early-diagnosis and drug-targed of diseases.Methods: HBV related fibrosis tissues and normal liver tissues were examined by two-dimensional gel electrophoresis (2-DE) after labeled with DIGE fluors Cy3, Cy5 and Cy2. Intensity changes of protein spots detected with statistical significance were identified by MALDI-TOF MS or MS/MS. To evaluate the reliability and accurate of the proteomic results, we used immunohistochemical technique to detect expression of some differently expressed proteins in the development of HBV related hepatic fibrosis tussues.Results: Intensity changes of 26 spots were detected with statistical significance. 15 protein spots of them up-regulated otherwise 11 down-regulated. A total of 19 different proteins were identified by MALDI-TOF MS or MS/MS successfully. There were 12 proteins including cytokeratin 8, shock protein 70 and high-mobility group box 1 up-regulation in HBV related hepatic tissues and other 7 proteins including thioredoxin peroxidase, ATP synthase and neuropolypeptide h3 down- regulation in HBV related hepatic tissues. These different proteins refer to oxidative stress, molecular chaperones, cystoskeleton, energy metabolism and signal transduction. Verifing the expression of the HSP70, CK8 and HMGB1 by using immunohistochemistry, it indecated that these proteins in hepatic fibrosis tissues were higher than that in normal liver tissues and the similar results were with proteomic analysis.Conclusions: Our results suggest that DIGE is a usfull technique for screening and identifying differential expressed proteins in HBV related fibrisis tissues. Through screening and identified related proteins of HBV related fibrosis, it may be useful for the exploration of new molecular markers for early-diagnosis and drug-targed of diseases. It also may provide a new system for study of the pathogenesis mechanism of HBV related hepatic fibrosis.