AP-1活化在高糖诱导肾小管上皮细胞转分化中的作用  

Effect of AP-1 on high glucose induced epithelial-mesenchymal-transition of human kidney proximal tubular epithelial cell

作　　者：吕智美[1] 邓华聪[1] 诸葛福媛[1] 刘金波[1] 陈丹燕[1] 南静[1] 

机构地区：[1]重庆医科大学附属第一医院内分泌科,重庆400016

出　　处：《第三军医大学学报》2008年第22期2107-2110,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30570414;30771038;30370670);教育部博士培养基金(20050631009);教育部第二批"春晖计划"(KJ050314);重庆市教委基金(2003-7)~~

摘　　要：目的探讨转录活化蛋白-1(activator protein-1,AP-1)在高糖诱导的肾小管上皮细胞表型转化中的作用。方法将体外培养的人肾小管上皮细胞(human kidney proximal tubular epithelial cell,HK2)分为正常对照组、高糖组、AP-1阻断组。光镜、透射电镜观察细胞形态的改变,免疫细胞化学法检测α-平滑肌肌动蛋白(α-SMA)和细胞角蛋白(cytokera-tin,CK)的表达,用凝胶电泳迁移率改变分析法(electrophoretic mobility shift assay,EMSA)检测HK2细胞AP-1的改变,RT-PCR法检测CK mRNA的表达,Western blot检测α-SMA蛋白的表达。结果高糖作用48h后,HK2细胞由椭圆形贴壁生长变为长梭形,胞体拉长,透射电镜下,细胞表面微绒毛明显减少,胞质内粗面内质网丰富,而AP-1阻断组较高糖组明显减轻;EMSA结果分析表明,在高糖刺激下,AP-1结合活力较正常对照组增加79.22%(P<0.05),而AP-1阻断剂可明显抑制AP-1的活化,较高糖组降低51.19%(P<0.05);免疫细胞化学和RT-PCR检测显示,高糖组和AP-1阻断组CKmR-NA和蛋白水平明显降低(P<0.05),而AP-1阻断组显著高于高糖组(P<0.05);免疫细胞化学和Western blot检测显示,高糖组和AP-1阻断组α-SMA蛋白表达明显高于正常组(P<0.05),而AP-1阻断组显著低于高糖组(P<0.05)。结论高糖能够导致肾小管上皮细胞AP-1活化,诱导其表型转化。Objective To examine the effect of AP-1 on high glucose induced epitheliat-mesenchymaltransition in cultured human kidney proximal tubular epithelial cell （HK2）. Methods The cultured HK2 cells were divided into three groups： normal glucose group （NG）, high glucose group （HG）, activator protein-1 （AP-1） inhibited group （AG）. The morphological changes of HK2 cells were observed by inverted microscope and electron microscope. The proteins of α-smooth muscle actin （α-SMA） and cytokeratin （CK） were determined by immunocytochemistry. The activity of activator protein-1 （ AP-1 ） was assessed with electrophoretic mobility shift assay （EMSA）. RT-PCR was used to detect the mRNA expression of CK. Western blot was used to detect the protein expression of α-SMA. Results The HK2 cells stimulated with high glucose became elongated and the number of microvilli decreased, but the number of rough endoplasmic reticulum increased. The level of AP-1 binding activity was increased by 79.22% after high glucose treatment, compared with that of normal glucose group, but decreased by 51.19% after the treatment of AP-1 inhibitor, compared with that in high glucose group. Cultured with high glucose, the expressions of CK mRNA and protein significantly decreased （P 〈 0. 05 ）, but they significantly increased in AP-1 inhibited group （P 〈 0.05）. Upon the stimulation of high glucose, the expression of α-SMA protein increased markedly （P 〈 0.05）, while it significantly decreased in AP-1 inhibited group （P 〈0.05）. Conclusion High glucose induces transdifferentiation of renal tubular cells probably through the activation of AP-1.

关 键 词：活化蛋白-1 肾小管上皮细胞 转分化 

分 类 号：R322.61[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]