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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:894  下载量:121
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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2017-09-25 09:55:23
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