p38MAPK介导高糖诱导的肾小管上皮细胞向间充质细胞转变解析.PDFVIP

生理学报 Acta Physiologica Sinica, December 25, 2008, 60 (6): 759-766 759 研究论文 p38 MAPK 介导高糖诱导的肾小管上皮细胞向间充质细胞转变 方开云，石明隽，肖瑛，桂华珍，郭兵，张国忠* 贵阳医学院病理生理学教研室，贵阳 550004 摘 要：本文旨在观察p38 MAPK 与高糖诱导的肾小管上皮细胞向间充质细胞转变之间的关系。将雄性Sprague-Dawley (SD)大鼠随机分为对照组、糖尿病组、胰岛素治疗组，用免疫组织化学、Western blot 检测p38 MAPK 和磷酸化p38 MAPK (p-p38 MAPK) 蛋白表达。采用机械分离和酶消化获取SD 大鼠肾小管节段，进行肾小管上皮细胞培养，将肾小管上皮细 胞分为对照组、高渗组(20 mmol/L D -mannitol) 、高糖组(20 mmol/L D-glucose)和SB202190 (p38 MAPK 特异性抑制剂) + 高 糖组，处理72 h 后收集细胞，用免疫细胞化学检测α- 平滑肌肌动蛋白(α-smooth muscle actin, α-SMA) 、p-p38 MAPK 和 Snail1 蛋白表达，Western blot 检测p38 MAPK 、p-p38 MAPK 、Snail1 、转化生长因子β1 (transforming growth factor-β1, TGF-β1)、α-SMA 和E-cadherin 的表达，RT-PCR 检测α-SMA 和E-cadherin mRNA 的表达。体内和体外结果均显示，高 糖状态激活了p38 MAPK ，这种活化作用在体内可因胰岛素控制血糖而被消除，在体外可被p38 MAPK 特异性抑制剂 SB202190 显著抑制；高糖组α-SMA 蛋白和mRNA 在原代培养肾小管上皮细胞的表达较对照组分别增加12 倍和8 倍(P0.01)， SB202190 处理组其表达则较高糖组分别减少67% 和50% (P0.01) 。SB202190 不影响TGF-β1 蛋白表达，但下调Snail1 蛋 白表达，并部分恢复高糖组E-cadherin 蛋白和mRNA 的表达。上述结果提示，p38 MAPK 可能通过转录因子Snail1 介导 高糖诱导的肾小管上皮细胞向间充质细胞转变。 关键词：p38 M APK ；上皮细胞向间充细胞转变；Snail1 ；糖尿病肾病；大鼠 中图分类号：R 3 2 9 p38 MAPK mediates high glucose-induced renal tubular epithelial-mesenchymal transition FANG Kai-Yun, SHI Ming-Juan, XIAO Ying, GUI Hua-Zhen, GUO Bing, ZHANG Guo-Zhong* Department of Pathophysiology, Guiyang Medical College, Guiyang 550004, China Abstract: The aim of the present study was to investigate the role of p38 MAPK in the renal tubular epithelial-mesenchymal transition (TEMT) induced by high glucose. In in vivo study, the rats were randomly divided into control (C), diabetes mellitus (DM) and insulin- treated DM groups. Immunohistochemical staining and Western blot were employed to determine the expression of p38 MAPK and p-p38 MAPK protein in renal cortex of rats. In in vitro study, primary renal tubular epithelial cells (PTECs) were cultured with normal gluc