法舒地尔对高糖培养人腹膜间皮细胞RhoA/Rock1、ROS、GSH-Px及TGF-β1表达的影响

Effect of fasudil on RhoA/Rock1,ROS,GSH-Px and TGF-β1 expression of mesothelial cells with high glucose condition

目的:探讨法舒地尔(FAS)对高糖培养的人腹膜间皮细胞(HMrSV)RhoA/Rock1、活性氧(ROS)、GSH-Px及转化生长因子β1(TGF-β1)表达的影响。方法:将人腹膜间皮细胞株HMrSV分为对照组(葡萄糖浓度为0)、高糖组(葡萄糖浓度分别为1.5%、2.5%和4.25%)和FSA干预组(2.5%葡萄糖+FAS 12.5、25和50μmol/L)。检测方法:Q-PCR法和Western印迹法检测RhoA和Rock1的表达;化学荧光法检测ROS的含量;ELISA法检测GSHPx和TGF-β1的含量。结果:与对照组相比,高糖组可呈浓度依赖方式上调RhoA、Rock1、ROS及TGF-β1的表达,下调GSH-Px的表达;与2.5%高塘组相比,FAS干预可明显下调RhoA、Rock1、ROS及TGF-β1的表达,上调GSH-Px的表达。结论:FAS可能通过抑制Rho激酶活性抑制高糖培养的人腹膜间皮细胞发生氧化应激反应。

Objective：To investigate the influence of fasudil on the expression of RhoA/Rockl, ROS, GSH-Px and TGF-131 of human peritoneal mesothelioma cell line （HMrSV） cells incubated with high glucose. Methodology： HMrSV were divided into control group, high glucose group （the concentrations of glucose were 1.5%, 2. 5% and 4. 25%） and fasudil treatment group （ glucose 2. 5% ＋ the concentrations of fasudil were 12. 5, 25 and 50 μmol/L） .The mRNA and protein expression levels of RhoA and Rockl were detected with Q-PCR and Western blot, the expression of ROS was detected with chemical fluorescence, the protein content of GSH-Px and TGF-β1 in the supernatant were detected with ELISA. Results：Compared with the control group, glucose could increase the expression of RhoA, Rockl, ROS and TGF-β, and down-regulate the expression of GSH-Px in a concentration dependent manner （P〈0. 05）. Fasudil could downregulate the expression of RhoA, Rockl, ROS and TGF-β1, and increase the expression of GSH-Px （P〈 0. 05）. Conclusion：Fasudil can inhibit high glucose-induced oxidative stress response of HMrSV cells and it may produce effect through the inhibition of Rho kinase activity.