Effects Of ADPN On Proliferation And Oxidative Stress Level Of HMCs Cultured With D-glucose Of High Concentration

Objective By cultivating human glomerular mesangial cells (HMCs) after intervention, observe high glucose and the added exogenous adiponectin in different time points, the HMCs proliferation, apoptosis, and differences in the levels of oxidative stress, realize that adiponectin could exert its protective effect by lowering sugar envionmenl oxidative stress levels in the cell, reveal the adiponectin has a very broad prospects in the field of prevention and cure of diabetes kidney disease.Methods (1)Test set up three groups, give HMCs different conditions for24h,48h,72h. N group (normal control group):the medium D-glucose concentration is5.5mmol/1; group G (high glucose):D-glucose concentration is30.0mmol/1; Adiponectin intervention group (group A):on the basis of the group G join concentration2.5ug/ml of adiponectin②At24h,48h,72h points each group was determined by MTT (methylazo thiazole blue) method for determining the light absorption (OD value), then observe the changes of cell proliferation in different conditions③Using the method of enzyme-linked immunosorbent (ELISA) determination of24h,48h,72h in each cell supernatant, the content of Caspase-3in observation of cell apoptosis.④Within24h,48h,72h, three time points using the AnnexinV/PI staining method, each cell apoptosis rate was measured by FCM.⑤Using fluorescence probe detection of groups of cells at different time points of reactive oxygen species (ROS), in order to understand oxidative stress levels in the cell.Results①Compared with normal control group, HMCs was inhibited in24h、48h、72h(P<0.05), but the proliferation of HMCs was increased apparently after ADPN being added(P<0.05).②he apoptotic of HMCs increased in high glucose group (P<0.05), but delayed in2.5μg/ml adiponectin group (P<0.05).③High glucose group compared with normal control group, the apoptosis rate of HMCs was obviously increased, after joining adiponectin intervention, HMCs apoptotic rate decreased, obvious difference (P<0.05).④High glucose group compared with normal control group, the FI value significantly increased, indicating the formation of intracellular reactive oxygen species significantly increased intracellular oxidative stress levels (P<0.05). after joining adiponectin, FI value declined obviously, close to the normal control group, the difference was statistically significant (P<0.05).Conclusion Long-term high glucose stimulating could inhibit cell proliferation of HMCs, but ADPN could promote the proliferation of the HMCs and protect them in D-glucose of high concentration. High sugar toxic effects can cause HMCs oxidative stress levels rise, beyond cells themselves antioxidant defense function, and cause cell injury. ADPN could protect HMCs in high density glucose. Adiponectin has hope become diabetic nephropathy early diagnosis and effective treatment of new directions.