黄芪甲苷对高糖诱导人肾小球系膜细胞损伤的保护作用及其机制  

Protective effects of As-IV on high glucose induced human mesangial cell injury and its mechanisms

作　　者：熊莉[1] 李维组 孙立[1] 李贵平[1] 李卫平[1,2] 

机构地区：[1]安徽医科大学药理学教研室,合肥230032 [2]安庆医药高等专科学校

出　　处：《安徽医科大学学报》2014年第9期1274-1278,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81173624);安徽省自然科学基金(编号:11040606M201);安徽省国际合作项目(编号:1230603007);安徽省教育厅自然科学基金(编号:KJ2012A192)

摘　　要：目的研究黄芪甲苷(As-IV)对高糖诱导人肾小球系膜细胞(HMC)损伤的保护作用及其机制。方法用HMC为目的细胞,实验分为正常对照组、甘露醇对照组、高糖组、Tempol(100μmol/L)组、转化生长因子-β(TGF-β)阻断剂SB431542(10μmol/L)组与As-IV(25、50、100μmol/L)组。各组细胞培养48 h后,用MTT法检测HMC增殖状况,二氢二氯荧光素(DCFH-DA)法检测细胞内活性氧(ROS)含量,ELISA法检测细胞上清液中IV型胶原(ColⅣ)的表达,Western blot法检测细胞中转化生长因子-β1(TGF-β1)、pSmad2/3、NADPH氧化酶4(NOX4)、瞬时受体电位阳离子通道蛋白6(TRPC6)蛋白的表达。结果与高糖组比较,Tempol组、SB431542组、As-IV 25、50、100μmol/L组均能显著抑制高糖诱导的细胞增殖,减少细胞内ROS及上清液中ColⅣ的含量,降低高糖诱导细胞内TGF-β1、p-Smad2/3、NOX4蛋白的高表达,并提高TRPC6蛋白的表达(P<0.05,P<0.01)。结论 As-IV对高糖诱导HMC损伤具有保护作用,其机制可能与抑制细胞增殖,减少细胞内ROS生成,下调TGF-β1、p-Smad2/3、NOX4蛋白的表达及上调TRPC6蛋白的表达有关。Objective To investigate the protective effects of astragaloside IV（As-IV） on high glucose induced hu- man mesangial cell （HMC） injury in vitro and its underlying mechanisms. Methods HMC was divided into eight groups and treated as follows： normal group （ NG）, mannitol （MA） , high glucose group （HG） , tempol （ 100 μmol/L）, SB431542 （10 μmol/L）, As-IV （25, 50, 100 p, mol/L）. After treatment for 48 hours ,the proliferation was analyzed by MTT assay. The reactive oxygen species （ROS） production was measured by 2, 7-dichlorodihydro fluorescin diacetate （DCFH-DA）. The level of Col IV in the supernatant was detected by ELISA. The expressions of transforming growth factor-βl （TGF-βl） ,p-Smad2/3, NADPH oxidase 4 （NOX4） and transient receptor poten- tial cation channel 6 （TRPC6） were analyzed by using Western blot. Results Compared with HG group, As-IV, Tempol, SB431542 treatment significantly inhibited HMC proliferation and overproduction of Col IV induced by high glucose. As-IV, Tempol and SB431542 treatment also decreased expression of TGF-βl, p-Smad2/3, and NOX4, but significantly increased expression of TRPC6 in HG-cultured HMC （ P 〈 0. 05 or P 〈 0. 01 ）. Conclu- sion As-IV has protective effects on high glucose induced HMC injury due to inhibit HMC proliferation, enhance anti-oxidant activity, suppress the expression of TGF-βl, p-Smad2/3, and NOX4 and up-regulation of TRPC6 ex- pression.

关 键 词：肾小球系膜细胞 糖尿病肾病 黄芪甲苷 TGF-β1 TRPC6 

分 类 号：R587.1[医药卫生—内分泌] R285.5[医药卫生—内科学]