基础医学与临床 ›› 2010, Vol. 30 ›› Issue (11): 1184-1187.

• 研究论文 • 上一篇    下一篇

AMPK在高糖诱导大鼠肾小球系膜细胞Ⅳ型胶原过度表达中的作用

贾振华 刘毅 曹铭锋 徐静 完强 王荣   

  1. 山东大学附属省立医院 山东大学附属省立医院
  • 收稿日期:2009-09-21 修回日期:2010-01-18 出版日期:2010-11-05 发布日期:2010-11-05
  • 通讯作者: 王荣

Role of AMPK in over-expression of collagen IV in rat glomerular mesangial cells induced by high glucose

Zhen-hua JIA, Yi LIU, Ming-feng CAO, Jing XU, Qiang WAN, Rong WANG   

  1. Provincial Hospital affiliated to Shandong University Provincial Hospital affiliated to Shandong University
  • Received:2009-09-21 Revised:2010-01-18 Online:2010-11-05 Published:2010-11-05
  • Contact: Rong WANG

摘要: 目的 观察高糖环境下大鼠肾小球系膜细胞Ⅳ型胶原蛋白表达水平的变化,以及这种变化与AMP活化蛋白激酶(AMPK)的关系。方法 实验分4组:对照组(糖浓度5.6 mmol/L),高糖组(22.0mmol/L),低浓度5-氨基-4-甲酰胺咪唑核糖核苷酸(AICAR)组(0.5mmol/L AICAR+高糖),高浓度AICAR组(1.0mmol/L AICAR+高糖)。孵育24h后,用RT-PCR法测定AMPKα1 mRNA水平,用Western blot法分别测定总AMPKα蛋白(AMPK)、磷酸化AMPKα蛋白(p-AMPK)和Ⅳ型胶原蛋白α5(COL4α5)亚单位的表达水平。结果 高糖组AMPKα1 mRNA水平低于对照组(p<0.01);高糖组总AMPKα蛋白和磷酸化AMPKα蛋白表达水平均低于对照组(p<0.01),高糖组COL4α5亚单位表达水平高于对照组(p<0.01);AMPK激活剂AICAR可在不同程度上纠正上述变化。结论 高糖环境下大鼠肾小球系膜细胞Ⅳ型胶原蛋白表达水平上调,这可能与AMPK表达下调及其活性降低有关。

关键词: AMP活化蛋白激酶, 高糖, 系膜细胞, 细胞外基质, Ⅳ型胶原蛋白

Abstract: Objective To observe the expression of collagen IV (COL IV) in cultured rat glomerular mesangial cells (GMCs), and to investigate the role of AMP-activated protein kinase (AMPK) in the process. Methods GMCs were incubated in four groups for 24 hours: control group (glucose 5.6 mmol/L), high glucose group (glucose 22.0mmol/L), low 5-amino-4-imidazolecarboxamide-riboside (AICAR, activator of AMPK) group (0.5mmol/L AICAR+22.0mmol/L glucose), high AICAR group (1.0mmol/L AICAR+22.0mmol/L glucose). AMPKα1 mRNA level was measured by RT-PCR, and the protein levels of AMPKα, p-AMPKα and COL4α5 subunit by Western blot analysis. Results Compared with control group, high glucose increased COL4α5 synthesis of GMCs, but reduced AMPKα1 mRNA level and protein levels of AMPKα, p-AMPKα; AICAR suppressed the over-expression of COL4α5, and raised the protein levels of AMPKα, p-AMPKα. Conclusion High glucose can increase COL4α5 synthesis of GMCs, this change maybe correlates with lower expression and activity of AMPK.

Key words: AMPK, high glucose, glomerular mesangial cells, extracellular matrix, collagen IV