整合素连接激酶在肾小管上皮细胞转分化中的作用  

Role of integrin-linked kinase in renal tubular epithelial-to-mesenchymal transition

作　　者：林琳[1] 张桦[1] 张慧涛[1] 贾宁[1] 汪悠悠[1] 朱晔[1] 周文英[2] 彭晓[2] 

机构地区：[1]中山大学附属第五医院肾内科,广东珠海519000 [2]中山大学附属第五医院中心实验室,广东珠海519000

出　　处：《中华临床医师杂志（电子版）》2009年第7期57-61,共5页Chinese Journal of Clinicians(Electronic Edition)

基　　金：广东省珠海市科技计划项目(PC20061077)

摘　　要：目的探讨整合素连接激酶(ILK)在转化生长因子β1(TGF-β1)介导的肾小管上皮细胞转分化(EMT)中的表达及作用。方法采用不同浓度的TGF-β1刺激体外培养的人肾小管上皮细胞(HKC),观察HKC形态学改变,并应用Western blotting方法检测其ILK、α-平滑肌肌动蛋白(α-SMA)的表达,实时RT-PCR法测定ILK、α-SMA、纤维连接蛋白(FN)、E钙黏蛋白(E-cadherin)mRNA的表达。结果(1)细胞形态学改变:在TGF-β1刺激下HKC细胞由正常状态下的圆形或卵圆形变为梭形,且随着孵育时间的延长,梭形细胞比例增大。(2)Western blotting和实时RT-PCR检测结果:在TGF-β1刺激下,ILK和α-SMA的蛋白表达量显著增加;ILK、α-SMA和FN的mRNA表达量也明显升高,而E-cadherin mRNA表达量明显减少。TGF-β1呈时间和剂量依赖性刺激HKC表达ILK蛋白和mRNA。(3)相关分析:ILK蛋白表达量与α-SMA蛋白表达量呈正相关(r=0.940,P<0.01);ILK mRNA表达量与α-SMA mRNA、FN mRNA表达量呈正相关(r=0.926,r=0.915,P<0.01),与E-cadherin mRNA表达量呈负相关(r=-0.820,P<0.01)。结论ILK参与TGF-β1诱导的肾小管EMT,并在其中发挥关键作用。Objective To investigate the expression and role of integrin-linked kinase （ILK） in the renal tubular epithelial to mesenchymal transition （EMT） induced by TGF-β1. Methods Human kidney proximal tubular cell line （HKC） was cultured in vitro. Cells were then treated with TGF-β1 at different concentrations. ILK, α-smooth muscle actin （ α-SMA ）, fibronectin （ FN ） and E-cadherin expression in protein and mRNA level were detected by Western blotting and real-time RT-PCR respectively. Results （ 1 ） Western blot analysis revealed that TGF-β1 significantly induced ILK and α-SMA protein expression in HKC cells in a dose- and time-dependent manner. Real-time RT-PCR analysis illustrated that expression for ILK mRNA,α-SMA mRNA, and FN mRNA were significantly increased, while E-cadherin mRNA was markedly decreased in a dose- and time-dependent manner in HKC cells after stimulation by TGF-β1. （2）Correlation analysis：expression of ILK protein was positively correlated with α-SMA protein （ r = 0. 940, P 〈 0. 01 ） ; expression of ILK mRNA was positively correlated with expression of α-SMA mRNA and FN mRNA （ r = 0.926,r = 0.915, P 〈 0.01 ）,while was negatively correlated with the E-cadherin mRNA expression （r = -0. 820,P 〈0. 01 ）. Conclusions ILK plays a crucial role in regulating TGF-β1 mediated renal tubular EMT.

关 键 词：转化生长因子Β1 细胞转分化 信号传导 整合素连接激酶 

分 类 号：R692[医药卫生—泌尿科学]