Proteomic Research On Rat Liver Fibrosis Of Decellularization Scaffold

[Objective]With the progress of the tissue engineering,using the proteomics research methods by two-dimensional electrophoresis?2DE?electrophoresis and LC-MS/MS techniques to compare the CCl₄ induced liver fibrosis in rats and normal rats model 2W,4W,6W and 8W group to cell biological scaffold protein,protein screening differences,through the formation process of the model of rat liver observation on the dynamic change of biological scaffold protein,biological scaffold for follow-up study to change the purpose of gene regulation,looking for diagnostic criteria and formulate reasonable classification experiment basis.[Method]Through the back subcutaneous injection of carbon tetrachloride(CCL_4,carbon tetrachloride)jointly a high-fat diet and ethanol induced the formation of SD rat liver tissue fibrosis,building period,a total of eight weeks.On the basis of the pathologic model,using the decellularizedl biological scaffold preparation technology,the liver fibrosis in rats model of 2W,4W,6W and8W with normal rats OW for proteomics research.Using HE and Masson staining to observe the histological changes of liver tissue and serological liver function tests further on liver fibrosis model identification.Followed by HE and Masson staining to observe the histological and ultrastructural changes of biological scaffold,immunofluorescence combined with 4',6-2amidine base-2-phenyl indole observe 5 groups of collagen?and?,Laminin?LN,Laminin?protein and fiber connection?FN,Fibronectin?observation of Extracellular Matrix?ECM,Extracellular Matrix?is the main component of DNA concentration residual DNA in qualitative and quantitative analysis of the organization and the fragment length,Elisa for ECM for detecting residues of various cytokines in the identification of liver fibrosis in preparation of biological scaffold;Finally using LC-MS/MS technology biological scaffold of 5 groups rats mass spectrum identification of the protein expression spectrum,to quantitative appraisal to the protein,in accordance with the quantitative 5 groups within the peptide peak area intensity?Indensity based absolute quantification,or iBAQ?^[1]the expression ratio of values between the specific size evaluation,summarizes the differences in liver fibrosis scaffold protein expression profile.GO?Gene Ontology?function classification analysis and KEGG?kyoko Engoolpedia of Genes and Genomes?signaling pathway enrichment analysis method to identify the differences in protein expression analysis.[Result]?1?Using CCL₄ is successful methods to building liver fibrosis model in rat,?2?using Triton X-100 unite NaOH is successful methods to building liver fibrosis biological scaffolds,?3?the formation of liver fibrosis is associated closely with four signal-channel which the up-regulated expression proteins?CYP4A10,CYP2C12 and CYP2C6?are mediated linoleic acid metabolism and steroid hormone biosynthesis,retinol metabolism and arachidonic acid metabolism,inflammatory mediator regulation TRP channels with retinol metabolism,arachidonic acid metabolism.[Conclusion]Formation of liver fibrosis is a complex process of signal transduction.Three proteins?CYP4A10,CYP2C12 and CYP2C6?are associated closely with liver fibrosis,and they way play critical roles in the process of liver fibrosis.