中文摘要：

目的研究黄芪甲苷（As-IV）对高胰岛素诱导人肾小球系膜细胞（HMCs）损伤的保护作用及其机制。方法以HMCs为研究对象,用MTT法检测不同浓度胰岛素作用不同时间后对HMCs增殖的影响,筛选胰岛素的量效与时效关系;实验设正常组、高胰岛素（64 nmol/L）组、二碘苯（DPI 10μmol/L）组、抗氧化剂Tempol（100μmol/L）组、PI3K/Akt信号通路抑制剂LY294002（10μmol/L）组与黄芪甲苷（As-Ⅳ25、50、100μmol/L）组。培养48 h后,MTT法检测HMCs的细胞增殖状况,DCFH-DA法测定细胞内活性氧（ROS）含量,Western blot法检测NADPH氧化酶4（NOX4）、磷酸化蛋白激酶B（p-PKB）（又称p-Akt）、磷酸化哺乳动物雷帕霉素靶蛋白（p-m TOR）蛋白表达的变化。结果随着浓度增加与作用时间延长,胰岛素对HMCs增殖有促进作用且有量效与时效关系;与高胰岛素组比较,DPI组、Tempol组、LY294002组与As-IV 25、50、100μmol/L组均能有效抑制高胰岛素诱导的细胞增殖,减少细胞内ROS含量,降低NOX4、p-Akt、pm TOR蛋白的高表达（P〈0.05,P〈0.01）。结论胰岛素呈时间和浓度依赖性促HMCs细胞增殖;As-IV对高胰岛素诱导的HMCs有保护作用,其作用机制可能与抑制HMCs的过度增殖,减少细胞内ROS生成,降低NOX4、p-Akt、p-m TOR蛋白高表达有关。

英文摘要：

Objective To investigate the protective effects of astragalosides IV（ As-IV） on high insulin induced human mesangial cells（ HMCs） injury in vitro and its mechanisms. Methods We used HMCs as research object,treated HMCs with different concentrations and time of insulin,analyzed the proliferation by MTT assay to select the dose-and time-dependent manners of insulin. The HMCs was divided and treated as follows： normal group（ NG）,high insulin group（ HINS,64 nmol / L）,DPI（ 10 μmol / L）,Tempol（ 100 μmol / L）,LY294002（ 10 μmol / L）and As-IV（ 25,50,100 μmol / L）. After treatment for 48 hours,the proliferation of HMCs was analyzed by MTT assay. The reactive oxygen species（ ROS） production was measured by 2,7-dichlorodihydto fluorescin diacetate（ DCFH-DA）. The expressions of NADPH oxidase 4（ NOX4）,phosphor-protein kinase b（ p-PKB） also named pAkt and phosphor-mammalian target of rapamycin（ p-m TOR） were analyzed by Western blot. Results The proliferation of HMCs was enhanced in a dose-and time-dependent manner by increasing concentrations and time of insulin. Compared with HINS group,DPI,Tempol,LY294002,As-IV treatment significantly inhibited HMCs proliferation,reduced intracellular ROS contents and decreased expression of NOX4,p-Akt and p-m TOR（ P〈0. 05,P〈0. 01）. Conclusion Insulin promotes proliferation of HMCs in a time-and dose-dependent manner. As-IV has protective effects on high insulin induced HMCs injury due to inhibition of HMCs proliferation,oxidative damage,NOX4,p-Akt and p-m TOR expression.