肾间质纤维化是糖尿病肾病的重要特征之一.DOC

*AP-1活化在高糖诱导肾小管上皮细胞转分化中的作用△ 吕智美 邓华聪* 诸葛福媛 刘金波 陈丹燕 南静（重庆医科大学附属第一医院内分泌科 400016） [摘要] 目的：探讨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（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(CK)的表达，用凝胶电泳迁移率改变分析法（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%（P0.05），而AP-1阻断剂可明显抑制AP-1的活化，较高糖组降低51.19%（P0.05）；免疫细胞化学和RT-PCR检测显示，高糖组和AP-1阻断组CK mRNA和蛋白水平明显降低（P0.05），而AP-1阻断组显著高于高糖组（P0.05）；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（P0.05），而AP-1阻断组显著低于高糖组（P0.05）。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，AP-1在肾间质纤维化时的EMT过程中可能起了重要的调控作用。 [关键词] 活化蛋白-1；肾小管上皮细胞；转分化 The effect of AP-1 on high glucose induced epithelial- mesenchymal - transition of human renal tubular epithelial cell Lù Zhi-mei , Deng Hua-cong* ，ZhuGe Fu-yuan, Liu Jin-bo,Chen Dan-yan ,Nan Jing (Department of Endocrinology , The first Affiliate Hospital Of Chongqing University Of Medical Sciences，Chongqing 400016, China) [Abstract] Objective To examine the effect of AP-1 on high glucose induced epithelial- mesenchymal- transition in cultured human renal 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 cell morphological changes were observed by inverted microscope and Electronic microscopy；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 levels of AP-1 binding activity w