Separation And Identification Of Proteomics In Colonic Mucosa Of Intestinal Tract Sub-health People

Objective Comparing and identification the colonic mucosa proteomic profiles difference between intestinal tract sub-health constipation and the bowel does not shape people, providing theoretical basis for understanding the pathogenesis of intestinal mucosa in intestinal tract sub-health constipation and the bowel does not shape people.Method Twenty intestinal tract sub-health constipation people, twenty the bowel does not shape people and twenty healthy volunteers are enrolled in this study. Biopsies were taken from the colonic mucosa under colonoscopy. The samples were washed by cold saline water, which contained 0.1% PMSF, and frozen in liquid nitrogen immediately. For the proteomic assay, total proteins were extracted and appled to the two-dimensional gel electrophoresis (2-DE). Proteins on 2-DE were visualized by the Coomassie blue stain, and the gels were scanned and analyzed with the Image Master 2D Elite. Mass spectrometry and bioinformatics analysis were employed to identify the proteins which were differently expressed.Results 535 protein spots were identified in the gel of the healthy controls, and 501 in the people with constipation, and 324 protein spots can be matched in these two groups (matching rate 64.67%).There were 175 protein spots that were significantly differentially expressed between the two groups, and 20 spots were picked for identification. Among the 20 spots,17 proteins were identified by mass spectrometry. In those 17 proteins, seven were decreased significantly, while ten were increased significantly. The up-regulated proteins were Sialic acid synthase, Poly(rC)-binding protein 1, Glyceraldehyde 3 phosphate dehydrogenase GAPDH, Esterase D, Carbonic anhydrase I, Tryptase beta I, MAWD binding protein, Proteasome activator complex subunit 2, Phosphatidylethanolamine-binding protein I, Peptidyl-prolyl cis-trans isomerase A. The down-regulated proteins were Cytokeratin-8, Aldehydedehydrogenasel A1 ZNF658 protein, Thioredoxin like protein p46,β-actin, Thiosulfate sulfurtransferase, YWHAZ protein.517 protein spots were identified in the gel of the bowel does not shape.366 protein spots can be matched to the control group, and the mean matching rate was 70.80%. There are 150 protein spots were significantly differentially expressed between the two groups, and 10 spots were picked for identification. Among the 10 spots,8 proteins were identified by mass spectrometry. In those 8 proteins, three were increased significantly, while five were decreased. The up-regulated proteins were Carbonic anhydrase I, Annexin A5 and Cyclophilin A. The down-regulated proteins were y-actin, Apolipoprotein A-I preproprotein, Aldehyde dehydrogenase 1A1 isoform 1,3-hydroxy-3-methylglutaryl-Coenzyme A synthase 2(mitochondrial) and Heat shock 70kDa protein 8.Conclusion 1.The standard, reproducible 2-DE patterns of intestinal tract sub-health constipation and the bowel does not shaped people are established.2. The 2-DE patterns of intestinal tract sub-health constipation group and control group were compared and 17 proteins were identified by mass spectrometry.10 proteins were increased significantly, while 7 proteins were decreased significantly. Among them, ZNF658 protein,β-actin, thiosulfate sulfurtransferase and YWHAZ protein may be associated with the pathogenesis of constipation; Aldehyde dehydrogenase 1, Peptidyl-prolyl cis-trans isomerase A, phosphatidylethanolamine-binding protein I and carbonic anhydrase I may be associated with the progress of long-term constipation to colorectal cancer.3.2-DE patterns of the bowel does not shaped group and control group were compared and 8 proteins were identified by mass spectrometry.3 proteins were increased, while 5 were decreased. Their function is under investigation.