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采用体外培养大鼠肾小球系膜细胞(rat mesangial cells,RMCs),MTT法检测RMCs增殖,DCFH-DA探针检测活性氧(ROS)生成,RT-PCR和Western blot检测铜锌超氧化物歧化酶(CuZnSOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(Gpx)、锰超氧化物歧化酶(MnSOD)和纤维连接蛋白(FN)mRNA和蛋白表达,探讨不同浓度通络方剂对高糖条件下大鼠肾小球系膜细胞(RMCs)体外增殖作用和氧化应激的影响。结果提示高糖条件下,通络方剂可抑制RMCs增殖(P<0.05),抑制活性氧生成(P<0.05),该抑制作用呈浓度依赖模式;高糖作用于RMC后,MnSOD和FN mRNA和蛋白水平升高,通络方剂可抑制高糖条件下FN mRNA的升高和提高正常糖浓度条件下MnSOD蛋白水平,浓度依赖性地抑制高糖条件下FN蛋白的表达。因此,通络方剂可抑制高糖条件下大鼠肾小球系膜细胞增殖,抑制氧自由基生成,提高抗氧化酶活性。
Rat mesangial cells (RMCs) were cultured in vitro. Proliferation of RMCs was detected by MTT assay. Reactive oxygen species (ROS) production was detected by DCFH-DA probe. The contents of copper and zinc superoxide (CuZnSOD), catalase (CAT), glutathione peroxidase (Gpx), manganese superoxide dismutase (MnSOD) and fibronectin (FN) Effect of Tongluo Prescription on Proliferation and Oxidative Stress of Rat Glomerular Mesangial Cells (RMCs) in High Glucose. The results indicated that Tonglu Prescription could inhibit the proliferation of RMCs (P <0.05) and inhibit the production of ROS (P <0.05) in a concentration-dependent manner under high glucose condition. After high glucose treatment, the expression of MnSOD and FN mRNA and Tongluo Recipe can inhibit the increase of FN mRNA under high glucose condition and increase the MnSOD protein level under normal glucose concentration, and inhibit the expression of FN protein under high glucose condition in a concentration-dependent manner. Therefore, Tongluo Prescription can inhibit the proliferation of rat glomerular mesangial cells in high glucose, inhibit the generation of oxygen free radicals and increase the activity of antioxidant enzymes.