摘要
目的观察巨噬细胞因子抵抗素过度表达对高糖刺激作用下人肾小球系膜细胞p38丝裂原活化蛋白激酶（MAPK）信号通路的影响，探讨抵抗素调控肾小球系膜细胞增殖及细胞外基质积聚的作用机制。方法通过转染携带野生型抵抗素基因的腺病毒载体（Ad—reSistin）构建过度表达抵抗素的人巨噬细胞模型，并与高糖刺激后的人肾小球系膜细胞共培养。3H-氚标胸腺嘧啶掺入法检测肾小球系膜细胞增殖。免疫细胞化学法检测系膜细胞激活蛋白1（AP-1）的表达。免疫荧光检测细胞外基质层粘连蛋白（LN）的表达。Western印迹检测系膜细胞内p38MAPK、转化生长因子（TGF）β1的表达并测定Smad2的磷酸化水平。结果Ad-resistin转染后，人巨噬细胞抵抗素mRNA水平及蛋白表达均显著升高（P〈0．01）。与对照组比较，与过度表达抵抗素的人巨噬细胞共培养后，人肾小球系膜细胞p38MAPK、TGF-β1的蛋白表达显著增强；细胞内Smad2的磷酸化水平显著升高（P〈0．05）；肾小球系膜细胞出现明显的增殖（P〈0．01）；细胞外基质的合成增多（P〈0．05）。结论巨噬细胞因子抵抗素的过度表达可能通过p38MAPK信号通路，促进高糖刺激作用下肾小球系膜细胞的增殖及细胞外基质的异常积聚。
Objective To investigate the effects of resistin on mesangial cells proliferation induced by high glucose and subsequent change of p38MAPK signal pathway. Methods Human macrophrages were cultured and treated with adenovirus encoding for resistin （Ad-resistin） for 48 h and were then co-cultured with human mesangial cells stimulated by high glucose for another 48 h. Mesangial cells were harvested and their proliferation was measured by 3H-TdR. Activator protein 1 （AP-1） was examined by immunocytochemistry and laminin of excellular matrix was observed with immuofluorescence. Protein levels of p38MAPK and TGF-131 were measured by Western blot. Smad2 phosphatase activity was aslo detected by Western blot. Results The mRNA and protein levels of resistin were significantly higher in Ad-resistin treated macrophages than those in Ad treated ceils （P〈0.01）. Over-expression of resistin up-regulated p38MAPK protein levels of human mesangial cells（P〈0.05）. Resistin also promoted the proliferation of mesangial cells （P〈0.01） and the synthesis of laminin stimulated by high glucose. The expression of TGF-β1 and phosphorylation of Smad2 were up-regulated in the mesangial cells （P〈 0.05）. Conclusion Macrophage cytokine resistin may promote mesangial cells proliferation and abnormal accumulation of excellular matrix stimulated by high glucose via activating p38MAPK signal passway.
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2008年第11期832-837,共6页
Chinese Journal of Nephrology
基金
重庆市自然科学基金（2006BB5074）
