Rac1抑制剂对高糖诱导大鼠肾小球系膜细胞损伤的影响及其作用机制

Effect of Rac1 inhibitor on high glucose induced injury of rat mesangial cells and its mechanism

目的探讨Rac1抑制剂(NSC23766)对高糖诱导大鼠肾小球系膜细胞损伤的影响及其机制。方法以大鼠肾小球系膜细胞为研究对象,将大鼠肾小球系膜细胞随机分为正常糖组(NG组)、高糖刺激组(HG组)、高糖刺激+NSC23766组(HG+NSC组)、溶剂对照组(HG+DMSO组)和渗透压对照甘露醇组(MG组)。应用CCK-8试剂盒检测不同浓度NSC23766对高糖诱导的大鼠肾小球系膜细胞活力的影响,采用流式细胞仪检测各组大鼠肾小球系膜细胞凋亡情况,采用Western blot法检测各组大鼠肾小球系膜细胞磷酸化氨基末端蛋白激酶(p-JNK)、Cleaved caspase-3蛋白表达水平、核因子κB(NF-κB)核转入水平,应用智能激光共聚焦显微镜分析各组细胞NF-κB表达情况。结果与NG组比较,HG组细胞活力降低(P<0.05),细胞凋亡率升高(P<0.05),p-JNK、Cleaved caspase-3蛋白表达量增加,NF-κB核转入增加(P<0.05);与HG组比较,HG+NSC(10μmol/L)组细胞活力明显增加(P<0.05),细胞凋亡率降低(P<0.05),p-JNK、Cleaved caspase-3蛋白表达量降低,NF-κB核转入减少(P<0.05);与HG+NSC(10μmol/L)组比较,HG+DMSO组细胞活力降低(P<0.05),细胞凋亡率升高(P<0.05),p-JNK、Cleaved caspase-3、蛋白表达水平增加,NF-κB核转入增加(P<0.05)。Rac1抑制剂(NSC23766)可使高糖诱导大鼠肾小球系膜细胞活力增加,凋亡率下降,降低p-JNK、Cleaved caspase-3蛋白表达水平和NF-κB核转入量。结论 NSC23766可通过抑制Rac1/JNK/NF-κB信号通路减轻高糖诱导的肾小球系膜细胞损伤。

Objective To investigate the effect of Rac1 inhibitor(NSC23766)on high glucose induced Rat glomerular mesangial cells injury and its mechanism.Methods Glomerular mesangial cells were used as the research object,rat mesangial cells were randomly divided into five groups:normal glucose group(NG group),high glucose stimulation group(HG group),high glucose stimulation+NSC23766 group(HG+NSC group),solvent control group(HG+DMSO group)and osmotic mannitol control group(MG group).The effect of different drug concentrations on high glucose induced mesangial cells injury was detected by CCK-8 kit.The apoptosis rate of rat mesangial cells was detected by flow cytometry.Western blot was used to detected the expression of phosphorylated amino terminal protein kinase(p-JNK)protein,nuclear factor-κB(NF-κB)nuclear transfer and Cleaved caspase-3 activation in glomerular mesangial cells of each group.The expression of NF-κB in each group was analyzed by intelligent laser confocal microscop.Results The cell viability of HG group was decreased,and the apoptosis rate,the expression of p-JNK,Cleaved caspase-3 protein,and the NF-κB nuclear transfer was increased compared with NG group(P<0.05).The cell viability of HG+NSC(10μmol/L)group was increased,and the apoptosis rate,the expression of p-JNK,Cleaved caspase-3 protein,and the NF-κB nuclear transfer was decreased compared with HG group(P<0.05).The cell viability of HG+DMSO group was decreased,and the apoptosis rate,the expression of p-JNK,cleaved caspase-3 protein,and the NF-κB nuclear transfer was increased compared with HG+NSC(10μmol/L)group.Rac1 inhibitor(NSC23766)attenuated high glucose induced glomerular mesangial cell injury in rats,decreased apoptosis rate,decreased expression of p-JNK and cleaved caspase-3 protein,and decreased NF-κB nuclear transduction.Conclusion NSC23766 can attenuate high glucose induced mesangial cell injury by inhibiting Rac1/JNK/NF-κB signaling pathway.