Proteomic Analysis Of Cocktail Therapy Liver Fibrosis In Rat

Objective:In the present study a rat model of CCl4-induced liver fibrosis was constructed, proteomic analysis of liver tissue was built up. The main purpose is to analysis differentially expressed proteins and clarify the molecular mechanism of cocktail therapy with taurine, epigallocatechin gallate and genistein against liver fibrosis, expecting to seek reliable biomarkers for the diagnosis,treatment and prognosis of liver fibrosis and to lay the foundation for the next experiment.Methods:(1) A rat model of liver fibrosis was constructed by CCl4. After six weeks’treatment of cocktail therapy, HE staining was performed to evaluate histopathologic changes. (2) Deplete the high-abundance proteins in tissue by ProteoMiner Protein Enrichment Kit and evaluate the effect of depletion by SDS-PAGE. The tissue of CCl4 model group, cocktail M and H group were labeled with iTRAQ reagents, then analyzed by SCX and LC-ESI-MS/MS quantitative proteomics. The resulting MS/MS spectra were searched against the rat sequence databases with MASCOT software, and analyzed by DAVID Functional Annotation Tool, STRING database and PATHWAY database.Results:(1) HE staining showed that cocktail therapy can alleviate the progression of liver fibrosis. (2) A total of 192 distinct proteins were identified. Among them,135 differential expressed proteins were identified in cocktail M group, of which 80 proteins were up-regulated and 55 were down-regulated.125 differential expressed proteins were identified in cocktail H group, of which 89 proteins were up-regulated and 36 were down-regulated. DAVID and STRING database analysis revealed that these proteins were involved in transcription, restructuring and signaling pathways, etc.Conclusion:Cocktail therapy can alleviate the progression of liver fibrosis effectively by affecting multiple biological processes, including glycolysis pathway, antioxidant defense system and coagulation cascade pathway. The molecular mechanism of cocktail therapy liver fibrosis may be through regulation of a variety of protein expression and activation or inhibit biological signaling pathways.