The Role Of FKBP51 In Mouse Liver Fibrosis And Its Molecular Mechanism

[Objectives] Liver fibrosis is the inevitable stage in the development of chronic liver disease to cirrhosis,which is the common pathological basis of chronic liver disease. Despite the high incidence of liver fibrosis,at present the pathogenesis is unclear and there is no effective anti-fibrosis treatment. Now the target treatment of TGF-beta pathway is rising, but the accurate anti-fibrosis mechanism needs further research.In this study we are going to analyze the phenotype of FKBP51 gene knockout mice to study the relationship between FKBP51 and liver fibrosis.【Methods】 1) Phenotype analysis. The experiment mice were divided into four groups: WT-Con (wild type mice injected with olive oil) s KO-Con (FKBP51 KO mice injected with olive oil)、 WT-CCL4 (wild type mice injected with olive oil and CCL4)、 KO-CCL4 (FKBP51 KO mice injected with olive oil and CCL4). Intraperitoneal injection three times per week for two weeks.2) Apparent pathological observation of liver.To observe the colour,the texture and the surface smoothness of the four group mice.3)Histopathological observation in liver.To study the histopathological change in liver, we do paraffin sections stained with hematoxylin and eosin (H&E),Masson staining and embedding in epon/araldite for electron microscopy of the four group mice.4) Real-time PCR.To test the molecule of liver fibrosis.5) Immunohistochemistry. This result can further validate the expression of liver fibrosis genes among the four group mice.6) Cell apoptosis.7) Western blot.Test the expression of some autophagy,lipid metabolism and TGF-βpathway related genes among the four group mice.【Results】 1) The animal model of hepatic fibrosis is built Successful with immunohistochemical method for testing.2) By Apparent pathological observation of liver, WT-CCL4 mice liver with dim colour,hard texture,surface roughness and full of small particles is more severe than KO-CCL4.3) Histopathological analysis of liver. Compared to WT-CCL4 mice, KO-CCL4 mice has less inflammatory cells and collagen deposition. 4)By Real-time PCR and Immunohistochemistry, we observe that the expression of fibrosis marker related protein Collagen Ⅰ,TIMP1,CTGF and a-SMA downregulated in KO-CCL4 mice than WT-CCL4,as well as cytokines CTGF、 IL6、 INF-Υ、 TGF-β、 TNF-α.5) Cell apoptosis. KO-CCL4 mice has less apoptotic cells than WT-CCL4.6) From Western blot,It was found that autophagy related genes increased in KO-CCL4 and the TGF-β pathway related genes Smad was activated.【Conclusions】 FKBP51 gene knock-out mice are resistant to CCL4-induced liver fibrosis.[Objective] The purpose of this study is to understand the function of Fkbp51 in metabolic pathways and neural pathways by profiling gene expression of liver and hippocampus tissues of both Fkbp51KO and WT mice.[Methods] mRNAs of liver and hippocampus of Fkbp51KO and WT mice were isolated and expression profiling was performed using RNA-seq. Differentially expressed gene between KO and WT were analyzed using BRB-Array Tools. DAVID, STRING, Genecard programs, the Gene Ontology, the Protein-protein Interaction Network and the Gene Annotation were applied to identify significant functional-relevant pathways.[Results] When compared differentially expressed genes between Fkbp51KO and WT in liver, we found that the loss of Fkbp51 in liver has large effect on the genes related to steroid biosynthetic and metabolic process, lipid biosynthetic process and oxidation reduction. When differentially expressed gene in hippocampus was studied between genotypes, we found that elimination of Fkbp51 has much effect on genes related to detection of mechanical stimulus, learning or memory, regulation of synaptic transmission and the pathway of PPAR and amyotrophic lateral sclerosis (ALS) e.g. When intersection of gene lists between liver and hippocampus tissues, we identified 11 common differentially expression genes. In these genes, HMGCS2 and INSIG1 are grouped in one protein-protein interaction network, and USP2, PER, CRY and DBP are grouped in another network.[Conclusions] The role of Fkbp51 in metabolism and nervous system is not only independent but also interactive.