黄芪甲苷对高胰岛素环境下肾小球系膜细胞的保护作用及其机制

Objective To research the protective effects of Astragalosides IV (AS-IV) on high insulin induced human mesangial cells (HMCs) injury in vitro and its potential mechanisdisms. Methods HMCs was taken as target cells. Treated HMCs with different concentrations and time of insulin, analyzed the proliferation and viability by MTT assay to select the dose-and time -dependent manners of insulin. The HMCs was divided and treated as follows: (1) normal group (NG), insulin (INS, respectively treated with 2, 4, 8, 16, 32, 64, 128 nmol/L insulin) group; (2) normal group (NG), high insulin group (HINS, 128 nmol/L), As-IV (25, 50, 100 μmol/L) and Tempol (100 μmol/L), LY294002 (10 μmol/L). After treating for 12 hours, the proliferation and viability was analyzed by MTT assay. The reactive oxygen species (ROS) producted in HMC was measured by DCFH-DA. Western blot was used to test the expression of p-Akt、p-gsk-3? and TRPC 6 protein. Flu-3 / AM as a fluorescent probe for detecting intracellular calcium ion concentration in each group. Results The proliferation of MHCs was enhanced in a dose-and time-dependent manners by increasing concentrations and time of insulin. Compared with HINS group, Tempol, LY294002, As-IV treatment significantly inhibited HMCs proliferation, reduced intracellular ROS contents and decreased the expression of p-Akt and p-gsk-3? ( P＜0.05，P＜0.01 ), increased the expression of TRPC6 and increased intracellular calcium ion concentration. Conclusion Insulin can promote the proliferation of HMC cultured in vito. As-IV has protective effects on HMCs induced by high insulin injury relate to inhibit HMCs proliferation, against oxidative damage, reduces p-Akt, p-gsk-3β protein content, increased expression of TRPC 6 and increased calcium ion concentration in cells.