Identification And Bioinformatics Analysis Of Differentially Expression Proteins In Rats With Renal Insufficiency Based On ITRAQ Technology

BackgroundRenal insufficiency is a group of clinical syndrome including glomerular damage,renal function decline,electrolyte and acid-base balance disorders,metabolic disorders that caused by various complex factors.Renal insufficiency is a disease with high incidence,high risk and severely impair the health and life of patients.At present,the mechanism of the occurrence and development of renal insufficiency is not perfect.We expect find out the differential proteins between the renal insufficiency and the normal rats by proteomics,and provide the experimental basis for the development of renal insufficiency.Objectivewe analysed differential proteins of renal insufficiency rats in the course of occurrence and progress by iTRAQ-based proteomics,and identified and verified these related proteins.Differentially expressed proteins were analyzed by bioinformatics analysis including GO analysis,KEGG analysis and STRING analysis on omicsbean software.MethodsThe 44 male SD rats(weight 180-220 g),which were randomly divided into control group(A),low-dose group(B),medium-dose group(C)and high-dose group(D)with 11 rats in each group.The rats in control group were fed normally,the rats in the other groups were fed with 20g/d forage containing adenine of 0.125%(B),0.25%(C),0.5%(D),respectively,for 21 day.On the last day of modeling,the body mass,24 h water intake and urine volume were measured,blood urea nitrogen(BUN)an serum creatinine(SCr)of rats were detected in each group.Histomorphology of glomerulus was detected by HE staining,differentially expression proteins were identified by ITRAQ-based proteomics.The expressions and levels of m RNA of haptoglobin,pentaxin-3,serotransferrin expression were verified by real-time quantitative PCR,Western blotting and enzyme-linked immunosorbent assay.ResultsThe renal insufficiency rats were established and the rats in each model group showed the different symptoms in different periods of renal insufficiency with each dose of adenine.The serum levels of Scr and BUN were increased in different degrees,renal tissues showed different degrees of damage by HE staining.The overall proteins were compared among the four groups.In total,416 proteins were confirmed.29 proteins withdifferential expression were found using stringent criteria.,we found that the 29 proteins mainly distributed in the extracellular region,blood particles and membrane bounded vesicles,function in molecular function regulation and enzyme regulator activity etc,involved in inflammation and response to external stimuli such as biology by omicsbean online software.we selected 3 proteins including HP,PTX 3 and TRF,from the 29 differentially expressed proteins by analyzing the results and reviewing the literature.The expression levels of the 3 proteins were consistent with the results of mass spectrometry by western blot,RT-PCR and ELISA.ConclusionWe have successfully established the rat model of renal insufficiency at different stages,and screened some differentially expression proteins based on iTRAQ technology,these proteins were analyzed by bioinformatics,we found that the occurrence and progress of renal insufficiency is related to multiple proteins,biological process,protein pathway and protein regulation network.It is suggested that HP,PTX3 and TRF may be related to the progression of renal insufficiency,which provides some experimental basis for the treatment of renal insufficiency.