Study On The Protective Effect Of Carnosine On Methylglyoxal Injury To HK2 Cells And Its Mechanism

Objective:To study the damage of methylglyoxal on HK2 cells and the protective effect of camosine on HK2 cells,and to further explore the protective mechanism of camosine on MGO-injured renal tubular epithelial cells through iTRAQ mass spectrometry analysis.Use hioinformatics analysis to find possible involvement of MGO and camosine Biological process,providing experimental evidence for new targets for the treatment of diabetic nephropathy.Methods:In this study,we designed to use camosine to interfere with MGO-induced renal tubular epithelial cells.The experiments were divided into control group,MGO group,camosine group,and MGO plus camosine group.Each group was set up with three biological replicates,and a series of experiments were performed after 48 hours of culture.The CCK-E method was used to detect the effects of MGO and camosine on the proliferation of HK2 cells.The cell scratch test was used to detect the effects of MGO and camosine on the migration of HK2 cells.The TUNEL test was used to detect the effects of MGO and camosine on HK2 cell apoptosis.Effects of MGO and Camosine on Apoptosis of HK2 Cells.In addition,iTRAQ mass spectrometry was used to analyze the expression of differential proteins,and bioinformatics analysis was used to find the biological processes that MGO and camosine may participate in,and to further explore the protective mechanism of camosine on MGO-induced renal tubular epithelial cells.Results:(1)CCK8 experiments showed that MGO at different concentrations(400,800,1200?M)all inhibited the proliferation of HK2 cells,and the optimal concentration was 800?M.Camosine can antagonize the toxic effect of MGO on HK2 cells,enhance cell viability,and the maximum protective effect concentration is 60 mM.(2)TUNEL apoptosis experiments showed that MGO promotes apoptosis of HK2 cells,while camosine can inhibit MGO-induced apoptosis.(3)Cell scratch test showed that MGO significantly inhibited the migration ability of HK2 cells,and the addition of camosine could enhance its migration ability.(4)Western blot showed that MGO up-regulated the expression of the apoptosis proteins Caspase3 and Bax in HK2 cell,and down-regulated the expression of camosine.(5)iTRAQ protein profile analysis MGO and camosine can significantly change the protein expression profile of HK2 cells.Conclusion:Camosine can protect HK2 cells from injury induced by methylglyoxal,and iTRAQ profiling analysis of MGO and camosine can significantly change the protein expression profile of HK2 cells.These findings provide new insights into the effects of MGO on proximal tubule cells in the kidney and help to better understand the mechanism by which camosine protects diabetic nephropathy.