Effects Of Adiponectin On Apoptosis And Oxidative Stress In Humanrenal Tubular Epithelial Cell Cultured With High Concentration Of D-glucose

Objective:This essay is intended to investigate the effects of adiponectin (ADPN) on apoptosis and oxidative stress in human renal tubular epithelial cells cultured with high concentration of D-glucose in vitro, and reveal the protective mechanism of adiponectin on diabetic nephropathy.Methods:①The human renal tubular epithelial cells were cultured with normal control group(5.5mmol/L D-glucose), high glucose group(30.0mmol/L D-glucose), high glucose plus adiponectin group(30.0mmol/L D-glucose+2.5μg/ml adiponectin). After being cultured for24h,48h,72h in each group, flow cytometry were designed to detect cellular apoptosis.②The human renal tubular epithelial cells were cultured with normal control group(5.5mmol/L D-glucose), high glucose group(30.0mmol/L D-glucose), high glucose plus adiponectin group(30.0mmol/L D-glucose+2.5μg/ml adiponectin). After being cultured for24h,48h,72h in each group, Superoxide dismutase (SOD) and maleic dialdehyde (MDA) in the supernatant of culture media were detected by xanthine oxidase method and thibabituric acid method.③The human renal tubular epithelial cells were cultured with normal control group(5.5mmol/L D-glucose), high glucose group(30.0mmol/L D-glucose), high glucose plus adiponectin group(30.0mmol/L D-glucose+2.5μg/ml adiponectin). After being cultured for24h,48h,72h in each group, Heme oxygenase-1(HO-1) mRNA and adaptor protein p66Shc mRNA expression were determined by RT-PCR.Results:①Compared with normal control group, the apoptosis rate of human renal tubular epithelial cells increased significantly in high glucose group, but decreased obviously with the treatment of adiponectin, showing the characteristic of time dependence.The difference has statistical significance(P<0.05).②Cmpared with normal control group, SOD activity decreased and MDA level increased in high glucose group, but SOD activity increased and MDA level decreased in adiponectin group, showing the characteristic of time dependence. The difference has statistical significance(P<0.05).③Compared with normal control group, the expression of HO-1mRNA and p66Shc mRNA both increased in high glucose group, but the expression of p66Shc mRNA decreased and HO-1mRNA continued to increase after adiponectin being added, showing the characteristic of time dependence. The difference has statistical significance(P<0.05).Conclusion:The high density D-glucose could induce HK-2cells apoptosis and oxidative stress. Adiponectin could delay HK-2cells apoptosis, increase SOD activity and the expression of HO-1mRNA, decrease MDA level and the expression of p66Shc mRNA. It might provide a new way for the clinical application of adiponectin in treatment of diabetic nephropathy.