螺内酯对高糖培养大鼠肾小球系膜细胞P-p38MAPK和TGF-β1变化的影响

Effect of high glucose and spironolactone on P-p38 mitogen-activated protein kinase and TGF-β1 in rat mesangial cells

目的:研究糖尿病肾病(diabetic nephropath,DN)大鼠肾小球系膜细胞p38丝裂原活化蛋白激酶(MAPK)的表达及螺内酯对其的影响。方法:以高糖孵育大鼠肾小球系膜细胞(mesangial rell,MC)24h,用细胞免疫荧光法检测MC的磷酸化p38MAPK(P-p38MAPK)活性,并用ELISA检测转化生长因子-β1(TGF-β1)分泌状况。p38MAPK特异性抑制剂SB203580及不同浓度螺内酯预处理对其影响。结果:高糖激活p38MAPK,增加RMC的P-p38MAPK活性和TGF-β1的表达;SB203580显著抑制TGF-β1表达(P<0.01);螺内酯抑制P-p38MAPK的活化并减少TGF-β1的蛋白表达(P<0.01),并随浓度增高,抑制作用增强。结论:p38MAPK可能是糖尿病肾病发生的始动信号之一,螺内酯可能通过抑制p38MAPK磷酸化而减少TGF-β1分泌。

Objective：To explore the effect of p38MAPK on diabetic nephropathy （DN） and the effect of spironolactone in rat.Methods：Rat mesangial cells（MC） were incubated with high glucose,after pre-treatment of SB203580 （p38MAPK special inhibitor） and spironolactone,the activity of P-p38MAPK was measured by immunofluorescence and TGF-β1 was examined by ELISA.Results：Compared with that of the control group,high glucose significantly activated p38MAPK and increased the activity of P-p38MAPK and the protein expression of TGF-β1 （P0.01）.The protein expression of TGF-β1 was significantly inhibited by SB203580 （P0.01）.Compared with those in MC incubated with high glucose,P-p38MAPK and TGF-β1 were expressed at a lower level in MC pre-treated with spironolactone （P0.01）.Spironolactone also decreased the P-p38MAPK and TGF-β1 expressions in MC at high glucose concentration in dose-dependent manner.Conclusion：p38MAPK may be one of the initial onset signals of diabetic nephropathy.Spironolactone may effectively attenuate DN progression through downregulating the expression of TGF-β1 via p38MAPK.