白藜芦醇对高糖诱导足细胞损伤的改善作用及机制探讨

Protective effect and mechanisms of resveratrol on hyperglycemia-induced podocyte apoptosis

关键词:  足细胞  细胞凋亡  白藜芦醇  氧化应激  线粒体功能障碍

Key words:  podocyte  cell apoptosis  resveratrol  oxidative stress  mitochondrial dysfunction

摘要:

[目的]观察白藜芦醇（RSV）对高糖诱导足细胞凋亡的影响及作用机制。[方法]采用体外高糖诱导作为足细胞损伤模型，细胞分为3组：对照组为5 mmol/L低糖培养液（LG）；模型组为30 mmol/L高糖培养液（HG）；RSV干预组：HG+5 μmol/L RSV；HG+10 μmol/L RSV，培养48 h后采用流式细胞术检测足细胞的凋亡情况；DHE探针观察细胞内活性氧自由基（ROS）生成；激光共聚焦检测线粒体膜电位及线粒体内ROS合成；Western blot检测线粒体凋亡途径相关蛋白含半胱氨酸的天冬氨酸蛋白水解酶（caspase）3、caspase 9的表达情况。[结果]与HG组相比，RSV可呈剂量依赖性的抑制高糖诱导的足细胞凋亡；荧光探针结果显示，RSV干预后细胞内及线粒体ROS生成明显低于HG组；JC-1染色显示，与HG组相比，RSV干预可显著提高线粒体膜电位；Western blot结果显示，随RSV干预浓度的提高，caspase 3、caspase 9表达水平明显下调，差异具有统计学意义（P<0.05）。[结论]RSV呈浓度依赖性的抑制高糖诱导的足细胞凋亡，其保护作用可能与抑制氧化应激，改善线粒体功能障碍相关。

Abstract:

[Objective] To observe the effect and mechanism of resveratrol (RSV) on podocyte apoptosis induced by hyperglycemia.[Methods] High glucose induction was used as a model of podocyte apoptosis in vitro. Podocytes were divided into three groups:Control group:5 mM D-glucose(LG);Model group:30 mM high glucose(HG);RSV intervention group:HG+5 μmol/L RSV;HG+10 μmol/L RSV. After podocyte cultured 48 h,the apoptosis rate was detected by flow cytometry. The intracellular ROS generation was evaluated using the dihydroethidium (DHE). Mitochondrial membrane potential andsuperoxide production of mitochondria were monitored with a confocal microscope. Western blot was used to observe the expression of caspase 3 and caspase 9,which were related with the mitochondria-mediated apoptotic pathway.[Results] Compared with the HG group,RSV protected against HG-induced podocyte apoptosis in a dose-dependent manner. The results of fluorescent probe showed intracellular and mitochondrial ROS generation after RSV intervention was lower than that in HG group. JC-1 staining showed that compared with HG group,RSV can significantly improve the mitochondrial membrane potential.Western blot results showed that compared with HG group. Cleaved caspase 9 and caspase 3 were significantly attenuated by RSV treatment in a dose-dependent manner. The difference was statistically significant(P<0.05).[Conclusion] RSV has the efficacy in protecting podocyte apoptosis induced by HG in a dose-dependent manner,and the mechanism may correlate with inhibiting oxidative stress and improving mitochondrial dysfunction.