黄芪甲苷的抗氧化应激作用对体外高糖刺激足细胞的保护作用

Effects of Astragaloside IV Protect Against Podocyte Lesion Exposed to High Glucose

目的:观察黄芪甲苷的抗氧化应激作用对体外高糖刺激下的足细胞的保护作用。方法:将条件性永生的小鼠足细胞随机分为正常对照组(NG)、高糖组(HG)、甘露醇高渗对照组(MA)、DMSO组及HG+不同剂量(5、15、30μg/ml)AS-IV干预组。采用CCK-8法检测各组足细胞活性,氧化应激试剂盒检测细胞中丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)水平;同时应用western印迹法检测细胞整合素连接激酶ILK蛋白表达量。结果:(1)足细胞活性:与NG组相比,MA组差异无统计学意义,HG组足细胞活性显著降低(P<0.05);与HG组相比,HG+30μg/mlAS-IV组足细胞活性显著升高(P<0.05),DMSO组、HG+15μg/mlAS-IV组和HG+5μg/mlAS-IV组差异无统计学意义(P<0.05)。(2)氧化应激及整合素连接激酶蛋白表达量:与NG组相比,HG组足细胞MDA含量升高,SOD、CAT和GSH-Px活性下降,ILK蛋白表达量下降(P<0.05)。与HG组相比,HG+30μg/mlAS-IV组MDA含量下降,SOD、CAT和GSH-Px活性升高,ILK蛋白表达量水平升高(P<0.05);HG+15μg/mlAS-IV组及HG+5μg/mlAS-IV组差异无统计学意义(P<0.05)。结论:黄芪甲苷(30μg/ml)的抗氧化应激作用对高糖诱导的足细胞损伤具有一定保护作用,可能与下调整合素连接激酶ILK系统有关。

Objective：The antioxidative effects of Astragaloside IV protect against podocyte lesion exposed to high glucose. Methods ： Conditionally immortalized mouse podocytes were randomly divided into 7 group： normal glucose group, high glucose group, mannitol group, DMSO group, HG ＋30 μg/ml AS- IV group, HG ＋ 15 μg/ml AS -IV group and HG ＋5μg/ml AS- IV group. Cell viability was examined by cell counting Kit - 8 （ CCK - 8） assay. Expression of ILK protein were observed by western blot. SOD , MDA, CAT and GSH -Px were measured by the assay kits, respectively. Results：Compared with incubated in NG, exposure to HG for 72 h significantly increase in levels of MDA and the expression of ILK, while a dramatical reduction in cell viability and activities of CAT, SOD and GSH- Px. However, HG- induced oxidative stress was partially reduced by AS -IV in a dose -dependent man- ner. No significant difference were found between NG group and MA group, as well as HG group and DMSO group. Conclusion：The antioxidative effect of AS - IV on podocyte injury under HG stimulation may be involved in ILK inhibition, and maybe a new treatment target in diabetic nephropathy.