The Effect Of High Glucose On Rat Tubular Epithelial Cells By Differential Proteomics Analysis

Objective To understand the effect of high glucose on rat renal tubular epithelial cellsprotein through observing their dynamic proteomics change under high glucoseenvironment, thereby hoping to find mechanisms for the development of diabeticnephropathy.Methods1As the tubular epithelial cells cultured in different based training, includingnormal glucose level （contain glucose5.5mM） and high glucose level（contain glucose30mM）. The total proteins were extracted after cells gathered at8hour,72hours,25days.2Thetotal proteins were separated using two-dimensional electrophoresis technology.3After theseparation gel application ImageMaster2D Platinum6.0on2-DE analysis software to analyzeimages, Evaluation by two-dimensional gel electrophoresis method was reproducible andreliability obtained by the unequal variance t-test p-value P<0.05volume%and the variation ofthe value ofthe point over two times as a standardto identifydifferentiallyexpressed proteins.4Different protein spots are identified by the technology of MS（MALDI-TOF, MALDI-TOF-TOF, LC-MS-MS）, and then the different protein is done bioinformatic analysis.5The resultusing the Mascot database. We judge the protein spots is based on the criterion that scores isgreater than60, sequence coverage is greater than15%, molecular weight （MW）discrepancy is less than30%and isoelectric point discrepancy is less than2.0between thecalculated value and the measured value. At last,the result revealed through searching thefunction ofthese proteins in PUBMEDdatabase.Results1Two-dimensional electrophoresis gel spots on average gained956.33±21.37spots,832points are present on all plastic, matching rate of87%.2There are8differential proteinspots between high glucose and normal at different time are found and are all identified（P<0.05）.:Moesin, annexin A4, voltage-dependent anion-selective channel protein2,phosphor glycerate mutase1, cellular nucleic acid-binding protein isoform2, stromal cell-derived factor2-like protein1precursor, capping protein, gelsolin-like, isoform CRA_b,nucleoside diphosphatekinase A.Conclusion Through the study of proteomics, We find out eight different proteins, whichindicates that the changes of these proteins is closely related to high glucose, It provides a newwayto studythe pathogenesis ofdiabetic nephropathy.