ILK介导TGF-β/Smad通路诱导肾小球内皮细胞-间充质细胞转分化的研究  

Effect of TGF-β/Smad signal pathway mediated by integrin linked kinase in the progress of renal fibrosis

作　　者：林琳[1] 郑晶[1] 朱伟平[1] 

机构地区：[1]中山大学附属第五医院肾内科,广东珠海519000

出　　处：《重庆医学》2015年第28期3911-3914,共4页Chongqing medicine

基　　金：珠海市科技计划项目(2013D0401990002)

摘　　要：目的观察TGF-β1在肾小球内皮细胞-间充质细胞转分化(EndMT)中的作用,探讨ILK介导TGF-β/Smad信号通路在EndMT过程中的作用。方法以体外培养人肾小球内皮细胞为研究对象,分为空白对照组、TGF-β1 12.5、25.0、50.0ng/mL组、TGF-β1 50.0ng/mL+Ⅰ型受体抑制剂(LY364749)5.0μmol/L组和ILK抑制剂(QLT-0267)5.0μmol/L组,各组细胞在上述处理因素下培养48、72h,以RT-PCR测定P-Smad2/3和ILK mRNA的表达水平,以Western blot测定P-Smad2/3、ILK及E-cadherin、CD31、α-SMA和FSP-1蛋白的表达水平。结果 (1)TGF-β1可剂量依赖性诱导HGEnCP-Smad2/3和ILK mRNA水平显著升高(P<0.01)以及P-Smad2/3、ILK、α-SMA和FSP-1蛋白水平显著升高(P<0.01),但是剂量依赖性诱导E-cadherin和CD31蛋白水平显著降低(P<0.01);(2)TGF-β1Ⅰ型受体抑制剂和ILK抑制剂可显著抑制TGF-β1诱导ILK mRNA水平的升高(P<0.01)以及ILK、α-SMA和FSP-1蛋白水平的升高(P<0.01),抑制E-cadherin和CD31蛋白水平的降低(P<0.01);(3)TGF-β1Ⅰ型受体抑制剂可显著抑制TGF-β1诱导P-Smad2/3 mRNA和蛋白水平的升高(P<0.01),但是ILK抑制剂对PSmad2/3mRNA和蛋白无显著影响(P>0.05)。结论 TGF-β1可促进肾小球内皮-间充质细胞转分化,ILK作为下游效应因子介导TGF-β/Smad信号通路参与了该过程,可能在肾脏纤维化过程中发挥重要作用。Objective To observe the effect of TGF-β1in the endothelial-mesenchymal transition EndMT of glomeruli,and to explore the effect of TGF-β/Smad signaling pathway mediated by integrin linked kinase（ILK）in the progress of renal fibrosis.Methods Human glomerular endothelial cells（HGEnC）incubated in vitro were divided into blank control group,TGF-β1（12.5,25.0,50.0ng/mL）group.TGF-β1 50.0ng/mL receptor type one antagonist（LY364749）5.0μmol/L group,ILK（QLT-0267）5.0μmol/L antagonist group.The mRNA level of P-Smad 2/3and ILK was determined by RT-PCR,and the protein level of P-Smad 2/3,ILK,E-cadherin,CD31,α-SMA and FSP-1was determined by Western blot after 48 hand 72hafter incubation in each group under the above-mentioned condition.Results（1）TGF-β1could significantly increased the mRNA level of P-Smad2/3and ILK（P〈0.01）,and the protein level of P-Smad2/3,ILK,E-cadherin,CD31,α-SMA as well as FSP-1（P〈0.01）in the concentration-dependent manner,but significantly decreased the protein level of E-cadherin and CD31（P〈0.01）in the concentration-dependent manner;（2）TGF-β1receptor type one antagonist and ILK antagonist significantly inhibited the increasing mRNA level of ILK（P〈0.01）,and the protein level of ILK,E-cadherin,CD31,α-SMA as well as FSP-1（P〈0.01）induced by TGF-β1in the concentrationdependent manner,and significantly inhibited the decreasing protein level of E-cadherin and CD31 induced by TGF-β1in the concentration-dependent manner（P〈0.01）;（3）TGF-β1receptor type one antagonist significantly inhibited the increasing mRNA and protein level of P-Smad2/3（P〈0.01）,but ILK antagonist had no effect to the increased mRNA and protein level of P-Smad2/3（P〈0.05）.Conclusion TGF-β1as the effector molecule in downstream can promote endothelial-mesenchymal transition of HGEnC,and TGF-β/Smad signaling pathway mediated integrin linked kinase participate in this process,which probably play important role in the progress of renal fibrosis.

关 键 词：整合素连接激酶 人肾小球内皮细胞 肾小球内皮细胞-间充质细胞转分化 转化生长因子-Β1 

分 类 号：R692[医药卫生—泌尿科学]