丝裂素活化蛋白激酶信号通路在人腹膜纤维化的作用  

Effect of MAPK signal transduction pathways on human peritoneal fibrosis

作　　者：樊敏[1] 段绍斌[1] 彭佑铭[1] 李军[1] 刘伏友[2] 

机构地区：[1]中南大学湘雅二医院肾内科中南大学肾病研究所,长沙410011 [2]中南大学湘雅二医院肾内科,长沙410011

出　　处：《中华肾脏病杂志》2003年第5期292-296,共5页Chinese Journal of Nephrology

基　　金：教育部高等学校重点实验室访问学者基金(2000170);湖南省自然科学基金(01JJY2071);湖南省卫生厅基金(2001-Y-39)

摘　　要：目的 探讨丝裂素活化蛋白激酶(MAPK)信号通路对转化生长因子(TGF)β1致人腹膜间皮细胞(HPMC)高表达纤连蛋白(FN)、纤溶酶原激活物抑制物(PAI)1的调控作用。方法 采用胰蛋白酶消化法从人腹膜组织中分离间皮细胞,建立稳定的体外培养模型。用四甲基偶氮唑盐比色法(MTT)评估TGF-β1和细胞外信号调节激素(ERK)及p38阻断剂对HPMC的增殖作用。采用酶联免疫双抗夹心法检测HPMC培养液中FN、PAI-1的蛋白质水平。采用逆转录多聚酶链反应(RT-PCR)检测HPMC细胞内FN、PAI-1mRNA的表达。结果 (1)TGF-β1能刺激HPMC增殖(P<0.05),且呈剂量时间依赖性,加ERK阻断剂组或p38阻断剂组较TGF-β1组有明显抑制作用(P<0.01)。(2)TGF-β1能引起HPMC FN、PAI-1蛋白水平显著增高(P<0.01);加ERK阻断剂组或p38阻断剂组能使上述高表达受到抑制(P<0.01)。(3)TGF-β1能使HPMC FN、PAI-1mRNA表达上调,加ERK阻断剂组或p38阻断剂组能使上调水平受到抑制。结论 MAPK对TGF-β1致人腹膜间皮细胞高表达FN和PAI-1起调控作用,为临床防治腹膜纤维化提供新的思路。Objective To explore the effect of transforming growth factor β1 (TGF-β1) on fibronectin (FN) and plasminogen activator inhibitor-1 (PAI-1) in cultured human peritoneal mesothelial cells(HPMC) and investigate the role of mitogen-activated protein kinase(MAPK) on TGF-β1 induced FN and PAI-1 in HPMC. Methods Mesothelial cells were isolated from human omental specimens by trypsin disaggregation and a stable HPMC culture model was established. The proliferation capacity of HPMC was assessed by tetrazolium salt colorimetry assay (MTT assay). ELISA was used to determine the quantity of FN and PAI-1 in culture supematants. Reverse transcription and polymerase chain reaction amplification was used to detect the expression of FNmRNA and PAI-1 mRNA in HPMC. Results (1) TGF-β1 could stimulate the proliferation of HPMC in a dose- and time-dependent mariner(P<0.05). (2)TGF-β1 remarkably increased the protein levels of FN and PAI-1 in the supematants(P<0. 01). (3) The expression of FNmRNA and PAI-1mRNA was up-regulated in HPMC exposured to TGF-β1.These above effects were suppressed by ERK inhibitor or P38 inhibitor. Conclusion TGF-β1 regulates FN and PAI-1 over expression in HPMC via MAPK signal transduction pathway,which leads to a new therapeutic way to peritoneal fibrosis.

关 键 词：丝裂素活化蛋白激酶 信号通路 腹膜纤维化 调控作用 转化生长因子Β1 腹膜透析 

分 类 号：R459.5[医药卫生—治疗学]