肝细胞生长因子对高糖刺激的系膜细胞GSK-3β和STAT3表达的影响  

Effect of hepatocyte growth factor on glycogen synthase kinase-3β and signal transducer and activator of transcription 3 expression induced by high glucose in rat mesangial cells

作　　者：李彩蓉[1] 蔡飞[2] 赵辛元[1] 杨颖乔[1] 余亚娟[1] 

机构地区：[1]咸宁学院临床医学院,湖北咸宁437100 [2]咸宁学院药学院,湖北咸宁437100

出　　处：《中国药理学通报》2010年第11期1439-1444,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No81000576);湖北省教育厅项目(NoB20092802;Q20082804)

摘　　要：目的观察肝细胞生长因子(HGF)对高糖环境下系膜细胞糖原合成酶激酶-3β(GSK-3β)和信号传导及活化转录因子3(STAT3)表达的影响。方法以大鼠系膜细胞为实验对象,给予HGF和高糖干预,实验中采用PI3K、p38MAPK、PKC、JAK2、ERK1/2抑制剂阻断其信号通路。系膜细胞转染野生型WT-GSK-3β,组成性激活型S9A-GSK-3β和显性负突变型KM-GSK-3β质粒,改变GSK-3β活性。Western印迹方法检测GSK-3β、pGSK-3β、STAT3、pSTAT3蛋白表达水平。酶联免疫吸附实验(ELISA)检测细胞培养液中单核细胞趋化因子(MCP-1)和细胞间黏附分子-1(ICAM-1)蛋白含量。结果 HGF能明显改善高糖刺激引起的磷酸化GSK-3β(Ser9)蛋白降低及抑制p-STAT3蛋白表达。c-Met抑制剂能完全抑制HGF对磷酸化GSK-3β蛋白的调节作用,PI3K、PKC、JAK2抑制剂能部分抑制HGF对磷酸化GSK-3β蛋白的调节作用,而p38MAPK和ERK1/2抑制剂对HGF的作用无影响。高表达S9A-GSK-3β则剥夺了HGF对pSTAT3蛋白的抑制效应,及降低炎症因子MCP-1、ICAM-1的表达。结论 HGF可通过HGF受体及PI3K、PKC、JAK2信号通路介导高糖诱导系膜细胞GSK-3β和STAT3蛋白磷酸化,HGF拮抗高糖环境下炎症因子表达可能与GSK-3β蛋白失活有关。Aim To study the effect of hepatocyte growth factor （HGF） on glycogen synthase kinase-3β （GSK-3β） and signal transducer and activator of tran- scription （ STAT3 ） of transcription expression induced by high glucose in rat mesangial cells. Methods High concentration glucose and HGF were used to stimulate the cultured rat mesangial cells in vitro. To investigate the direct effect of GSK-3β on mesangial cells, its activity was changed by transient transfection with three kinds of GSK-3β mutant plasmids, nonphosphorylatable constitutively active mutant of GSK-3β （S9A-GSK-3β）, catalytically inactive GSK-3β （KM- GSK-3β） and adenoviral vectors encoding wild-type （ WT- GSK-3β）. The protein expression of GSK-3β, pGSK-3β, STAT3, and pSTAT3 was determined by Western blot. The levels of monocyte chemoattractant protein-1 （MCP-1） and intercellular adhesion molecule-1 （ICAM-1） in the supernatants were determined by enzyme-linked immunosorbant assay （ELISA）. Results HGF increased the expression of pGSK-3β pro-tein and inhibited the expression of pSTAT3 protein, which was evident in mesangial cells induced by high glucose and HGF. C-Met inhibitor, completely inhibited the phosphorylation of GSK-3β protein, PI3K, PKC and JAK2 inhibitors could partially inhibit the phosphorylation of GSK-3β protein, while p38MAPK and ERK1/2 inhibitors had no effect. In contrast, ectopic expression of S9A- GSK-3β abolished the HGF suppressive action on STAT3, MCP-I and ICAM-1 protein expression. Conclusions HGF-induced downregulation of pSTAT3 involves activation of the c-Met, PI3K, PKC and JAK2 pathway and requires the activity of early GSK-3 β. These findings suggest that inhibitory phosphorylation of GSK-3β at Ser-9 is required for HGF inhibition of STAT3 in mesangial cells induced by high glucose.

关 键 词：肝细胞生长因子 糖原合成酶激酶-3Β 信号传导及活化转录因子3 单核细胞趋化因子 细胞间黏附分子-1 糖尿病肾病 

分 类 号：R-332[医药卫生] R329.24