整合素连接激酶介导的高糖诱导肾小管上皮细胞纤连蛋白的表达  

High-glucose up-regulates the expression of fibronectin mediated by integrin-linked kinase in renal tubular epithelial cells

作　　者：熊明霞[1] 谭若芸[1] 张飞飞[2] 王晓华[2] 方丽[2] 樊伟[2] 王笑云[1] 杨俊伟[1] 

机构地区：[1]南京医科大学附属第一医院肾内科江苏省糖尿病医学中心,210003 [2]南京医科大学附属第二医院肾脏病医学中心,210003

出　　处：《中华肾脏病杂志》2008年第8期571-574,共4页Chinese Journal of Nephrology

基　　金：国家自然科学基金（30270616,30470800）;科技部“973”重点项目（2006CB503909）;江苏省自然科学基金（BK20060403）;国家人事部回国人员科研启动基金;江苏省“六大人才”高峰项目（苏政办发[20011124号）;江苏省“科教兴卫”工程医学领军人才项目（苏卫科教[2007127号）

摘　　要：目的观察在高糖刺激下纤连蛋白（FN）与整合素连接激酶（ILK）在肾小管上皮细胞的表达情况，探讨糖尿病肾病小管间质纤维化的发病机制。方法以人肾小管上皮细胞株（HKC）细胞和链脲佐菌素（STZ）诱导的CD-1小鼠糖尿病动物模型为研究对象，采用Western印迹的方法检测细胞或肾组织ILK和FN蛋白的表达。通过基因转染的方法，将含无激酶活力的人ILK基因表达质粒pCMV-kdlLK转染HKC细胞，观察抑制ILK活力对高糖诱导的HKC合成FN的影响。结果STZ注射后4周，CD-1小鼠血糖水平显著高于对照组[（20．3±2．7）mmol／L比（6．1±1．4）mmol／L，P〈0．01]，同时，肾组织ILK和FN的表达量亦显著高于对照组，且ILK与FN表达量呈正相关（P〈0．01）。细胞培养实验证实，高糖能够刺激HKC上调ILK和FN蛋白的表达，并且呈时间和剂量依赖性。HKC细胞转染pCMV—kdILK质粒以抑制ILK的活力，能够显著地拮抗高糖刺激HKC表达FN的作用。结论高糖通过上调ILK蛋白增加肾小管表达并合成FN，抑制ILK能够拮抗高糖刺激HKC合成FN的作用。Objective To investigate the relationship between high-glucose-induced fibronectin（FN） expression and up-regulation of integrin-linked kinase（ILK） in human kidney tubular epithelial cells （HKC） and kidney of CD-1 mice. Methods Cultured human kidney tubular epithelial cells and streptozotocin （STZ）-induced diabetic model of CD-1 mice were enrolled in this study. Western blot was used to detect the expression of FN and ILK. The kinase dead ILK plasmid （pCMV-kdlLK） were transferred to HKC. Results Four weeks after injection of STZ, CD-1 mice had higher blood glucose level as compared to the control [（20.3±2.7） mmol/L vs （6.1±1.4） mmol/L, P〈0.01]. Meanwhile, expression of FN and ILK was significantly increased in diabetic mice as compared to the control （P〈0.01）. There was positive correlation between the expression of FN and ILK （r=0.899, P〈0.01）. High-glucose could up-regulate FN and ILK expression in cultured HKC in a time- and dose-dependent manner. Blockage of ILK activation by pCMV-kdlLK abrogated high-glucose-incuced FN expression in HKC. Conclusions Highglucose can induce FN expression through up-regulating ILK expression. Blockage of ILK activation abrogates this effect.

关 键 词：高糖 肾小管 整合素连接激酶 纤维连接蛋白 

分 类 号：R692.6[医药卫生—泌尿科学]