ITRAQ-based Proteomic Analysis Of Tuberculous Contracted Bladder & Stenosed Ureter And Differentially Expressed Protein Verification

Objective:In this study,proteomics techniques were used to screen for the differentially expressed protein between normal bladder and contracted bladder,normal ureter and stenosed ureter.Read the literature and screening of differentially expressed proteins that may be linked to bladder and ureteral fibrosis,development.Through the WB technique for differential protein validation,and provide new methods and targets for contracted bladder and stenosed ureter grading,diagnosisMethods:Part Ⅰ: Collected contracted bladder and stenosed ureter tissue which patient had been diagnosed as urinary tuberculosis in the PLA 309 hospital urology from January 2015 to December 2016;access to DCD donor bladder and ureter tissue as a normal control in our hospital in the same period;each group have 2 sample.After the four tissues were mixed,the total protein were analyzed by iTRAQ labeling technique combined with LC-MSMS.The proteins were identified as differentially expressed proteins respectively in normal bladder and contracted bladder,normal ureter and stenosed ureter tissues that over-expressed or under-expressed with 2times and more than 2 times.The common differentially expressed proteins was that both expressed in two kinds tissues.Use Uni Prot,David,string and Wego database to analyze those differential protein function,where its localization and interaction between other protein.Part II: selecting 3 blocks in four groups according to the preamble described.Each group go on Masson staining that can identify the collagen fibers expression in the ureteral tissues and bladder tissues.The expression of MFAP4 and FMOD were determined by WB which chose from common differentially proteins.Results:Part Ⅰ: A total of 188 differential proteins were identified between the normal bladder and contracted bladder and identified 130 different protein between the normal ureter and ureter stenosis.There are 48 kinds common differentially expressed proteins belong the two groups,including 20 kinds of proteins down regulated,28 up-regulated proteins.Among those 48 protein,there are 28 kinds of proteins were labeled in the David database.Those 28 proteins mainly involved in 17 types of biological process,5 types of molecular function and 13 kinds of cells;39 proteins were detected in the string database,three interaction networks were composed.Part II: Masson staining suggests that the collagen fibers were expressed muscular layer in contracted bladder and stenosed ureter.Contracted bladder versus normal bladder,stenosed ureter compared with normal ureter,collagen expression was significantly increased,the difference has statistical significance;Western blot results showed that MFAP4 and FMOD were significantly higher in the diseased bladder and ureter,and the difference was statistically significantConclusion:1、iTRAQ united LC-MSMS can qualitatively and quantitatively identify the protein and selected the differentially expressed protein,furthermore we can analysis its biology information;2 、The up regulation of FMOD in diseased tissues is an important factor affecting the synthesis of collagen fibers,and is closely related to the fibrosis of the bladder and ureter.3、MFAP4 is highly expressed in diseased tissues and can be used in a walking experiment to investigate its application in the diagnosis,grading and prognosis of tuberculous fibrosis...