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黄芪甲苷对高胰岛素环境下人肾小球系膜细胞的保护作用及其机制
发表时间：2017-09-15 浏览量：1964 下载量：443
| 全部作者： | 段文婷，李卫平，徐源，黄存东 |
| 作者单位： | 安徽医科大学药理学教研室；安庆医药高等专科学校 |
| 摘 要： | 目的：探究黄芪甲苷（astragaloside IV，AS-IV）对高胰岛素（high insulin，HINS）环境下人肾小球系膜细胞（human mesangial cell，HMC）损伤的调控及保护作用。方法：将HMC作为操作对象，设立如下分组：1）正常组（NG）、胰岛素组（INS）（2，4，8，16，32，64，128 nmol/L）；2）正常组（NG）、HINS组（128 nmol/L）、AS-IV组（25，50，100 μmol/L）、Tempol组（100 μmol/L）、LY294002组（10 μmol/L）。干预12 h后，用噻唑蓝（MTT）法检测HMC细胞状态与细胞数量的受影响情况；2’,7’-二氯荧光黄双乙酸盐（DCFH-DA）法检测每一组中活性氧（reactive oxygen species，ROS）表达量；Western blotting检测每一组HMC中总Akt、p-Akt、总gsk-3β、p-gsk-3β、TRPC6蛋白表达差异；Flu-3/AM荧光探针检测细胞内游离钙离子浓度。结果：不同时间和剂量下，胰岛素对细胞的增殖具有浓度依赖性促进作用；在实验剂量下，AS-IV对HINS环境下HMC的增殖具有随剂量递增的抑制作用。与HINS组相比，AS-IV可在一定程度上减少HMC内ROS含量，显著提高TRPC6蛋白的表达程度，下调p-Akt、p-gsk-3β蛋白表达程度，上升细胞内游离的钙离子浓度。结论：AS-IV对在HINS环境下HMC的生长具有保护作用，其原理可能与AS-IV提高HMC抗氧化能力，降低p-Akt、p-gsk-3β蛋白含量，提高TRPC6表达水平和细胞内游离钙离子浓度有关。 |
| 关 键 词： | 药理学；糖尿病；人肾小球系膜细胞；黄芪甲苷；钙离子；TRPC6 |
| Title： | Protective effects of AS-IV on high insulin-induced human mesangial cell injury and its mechanisms study |
| Author： | DUAN Wenting, LI Weiping, XU Yuan, HUANG Cundong |
| Organization： | Department of Pharmacology, Anhui Medical University; Anqing Medical College |
| Abstract： | Objective: To research the protective effects of astragalosides IV (AS-IV) on high insulin (HINS)-induced human mesangial cell (HMC) injury in vitro and its potential mechanism. Methods: HMC was taken as target cell. The HMCs were divided and treated as follows: 1) normal group (NG), insulin group (INS, respectively treated with 2, 4, 8, 16, 32, 64, 128 nmol/L insulin); 2) normal group (NG), high insulin group (HINS, 128 nmol/L), AS-IV group (25, 50, 100 μmol/L), Tempol group (100 μmol/L), LY294002 group (10 μmol/L). After treating for 12 h, the proliferation and viability were analyzed by MTT assay. The expression of reactive oxygen species (ROS) produced in HMC was measured by DCFH-DA method. Western blotting was used to test the expression of total Akt, p-Akt, total gsk-3β, p-gsk-3β and TRPC6 proteins. Flu-3/AM was used as a fluorescent probe for detecting intracellular calcium ion concentration in each group. Results: The proliferation of HMC was enhanced in dose- and time-dependent manners by increasing concentrations and time of insulin. AS-IV inhibited proliferation of HINS-induced HMC in a dose-dependent manner compared to HINS group, AS-IV treatments significantly inhibited HMC proliferation, reduced intracellular ROS contents and decreased the expression of p-Akt and p-gsk-3β, increased the expression of TRPC6 and intracellular calcium ion concentration. Conclusion: AS-IV has protective effects on HMC injury induced by HINS, which relate to increasing antioxidant ability of HMC, reducing p-Akt, p-gsk-3β protein contents, increasing expression level of TRPC6 and intracellular calcium ion concentration. |
| Key words： | pharmacology; diabetic mellitus; human mesangial cell; astragaloside IV; calcium ion; TRPC6 |
| 发表期数： | 2017年9月第17期 |
| 引用格式： | 段文婷，李卫平，徐源，等. 黄芪甲苷对高胰岛素环境下人肾小球系膜细胞的保护作用及其机制[J]. 中国科技论文在线精品论文，2017，10（17）：1919-1926. |
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