Quantitative Proteomics Study Of Nephritis Induced By HBs On HBs Transgenic Mouse

Hepatitis B virus(HBV)is a globally distributed infectious human pathogen,especially in sub-Saharan Africa,the Pacific,and especially Asia.The population living in the chronic high HBV prevalence rate accounts for 45% of the global population,and the population of HBs infected has reached 400 million,making HBV become a common human pathogen,seriously affecting social and economic development.HBV is a member of the hepadnavirus family.Chronic infection(Chronic HBV,CHB)causes a variety of liver diseases,including asymptomatic HBV carriers(ASC),chronic hepatitis(CH),and liver.Hardening(Liver Cirrhosis,LC)and hepatocellular carcinoma(HCC),etc.A variety of extrahepatic clinical manifestations have been observed in patients with acute and chronic liver disease infected with HBV.Renal dysfunction remains the most common extrahepatic clinical manifestation.This kidney disease caused by hepatitis B virus infection is called HBV-associated glomerulonephritis(HBV-GN).Its possible pathogenic mechanism hypothesis includes the typical immune complex glomerulonephritis mediated by HBV antigen(HBAg)and antibody(HBAb)immune complexes,defects in the body’s immune function caused by host and viral genetic factors,and autoimmune or viral direct effects on kidney organs,etc.Positive HBV-DNA could be detected in the nucleus of mesangial cells and renal tubular epithelial cells of HBV-GN patients,demonstrating the existence of a direct viral effector mechanism in the body.However,there are few studies on how to cause kidney disease after HBs directly affect the kidney,and the molecular mechanism is unclear.In this study,we used a stable HBs transgenic mouse(Mus musculus)as a model to explore the possible pathogenic mechanism of HBs by means of quantitative proteomics and weighted co-expressed bioinformatics network analysis of TMT method.wild-type mouse and HBs transgenic mouse at 6 and 12 months were labelled by TMT method,respectively.6 months wild-type mouse as biological replicates.All samples were mixed in equal amounts and then separated by offline liquid chromatography,and then the large-scale identification and quantification of protein were performed by LC-MS to observe the pathogenic mechanism of HBs and the difference in proteome caused by changes in infection time.We identified a total of 5127 proteins,the total number of MS2 was 665,687;the resolution of the MS2 was 23.4%.The proportion of the number of unique peptides greater than or equal to 2 is more than 85%.The Weighted Co-Expression Analysis Network(WGCNA)uses Pearson to calculate the correlation coefficient between the two proteins,and together the highly correlated proteins are defined as a module.The categorical variable,whether it is a wild type/transgenic sample,is converted into a numerical variable(0/1)for calculation.If the proteins that are clustered together in a module are highly correlated with the phenotype,then the protein of this module is very important.The differential protein was divided into three modules of brown,green and blue by means of a weighted co-expression analysis network algorithm.Correlation analysis was performed between the module and whether it was a transgenic infection phenotype.The green module was found to be significantly related to the phenotype.Among them,there were 127 significantly differential proteins associated with phenotypic.Further,the differential proteins were divided into 4 types as time of injection.The differential proteins were performed the enrichment analysis by Metascape,IPA and Reactome,this revealed three possible mechanisms of HBs transgene-induced nephropathy.(1)HBs effect the solute carrie(SLC)-related proteins which belongs to the down-regulated proteins,causing kidney damage;(2)HBs act as immunogens to activate the renal complement response,which in turn mediates the inflammatory pathway of the kidney;(3)HBs expression triggers apoptosis,releases cytochromes,and mediates abnormalities in PPAR pathway and inflammatory pathway.Not only that,but we also found five proteins with long-lasting expression of HBs,such as FABP1,PNKD,ECI3,PRL15 and HACL1.According to the literature,high expression of ribosomal protein L15(PRL15)is known to promote the proliferation of cancer cells;and liver fatty acid binding protein(hepatic fatty acid binding protein,L-FABP or FABP1)is associated with the development of chronic kidney disease,and can be used as tubulointerstitial Quality damage sensitive predictor.