巨噬细胞移动抑制因子经Rho途径促进人肺成纤维细胞Ⅰ型胶原合成  

Effect of Recombinant MIF on Lung Fibroblasts Collagen Synthesis in Vitro

作　　者：陈培芬[1] 罗雅玲[2] 赖文岩[3] 邢晓雯[4] 谭家余[2] 骆子义[1] 邱智辉[1] 

机构地区：[1]广东省深圳市第三人民医院内科,5180114 [2]南方医科大学附属南方医院呼吸科 [3]南方医科大学附属南方医院心内科实验室 [4]广州医学院附属第一医院心内科

出　　处：《医学理论与实践》2011年第14期1621-1624,共4页The Journal of Medical Theory and Practice

基　　金：国家自然科学基金(30770936)

摘　　要：目的:观察重组巨噬细胞移动抑制因子(rMIF)对人胚肺成纤维细胞(MRC-5)的作用,探讨哮喘气道重塑的发生机制。方法:100μg/L的rMIF刺激MRC-5细胞6h、12h、24h、48h后,ELISA法测定细胞培养上清中TGF-β1蛋白含量;100μg/L的rMIF作用于MRC-5细胞48h,RT-PCR法检测Ⅰ型胶原mRNA表达,Western blotting检测Ⅰ型胶原蛋白合成;100μg/L的rMIF刺激MRC-5细胞48h,刺激前0.5h加入Rho激酶特异性抑制剂Y27632,RT-PCR法检测Ⅰ型胶原mRNA表达,western blotting法检测Ⅰ型胶原蛋白表达。结果:与对照组比较,100μg/L的rMIF对细胞培养上清TGF-β1蛋白含量无显著影响(P>0.05);100μg/L的rMIF可显著促进Ⅰ型胶原mRNA(P<0.01)和蛋白(P<0.01)合成。Y27632预刺激后,100μg/L的rMIF促进Ⅰ型胶原mRNA和蛋白合成能力显著减弱(P均<0.01)。结论:MIF可能通过Rho途径刺激人肺成纤维细胞Ⅰ型胶原合成从而在哮喘气道重塑的发病机制中发挥重要作用。Objective：To investigate the influence and mechanism of recombinant macrophage migration inhibitory factor（rMIF） on fibroblasts.Methods：After 100μg/L rMIF challenged for 6,12,24 and 48h,the culture supernants of MRC-5 fibroblasts were obtained and TGF-β1 protein was detected by ELISA.Then the cells were divided into two groups：the treated group was treated with rMIF（100μg/L,48h）and the control was without rMIF treatment The expression of collagen type ⅠmRNA was examined using RT-PCR analysis.The level of collagen type Ⅰ protein induced by rMIF was quantified by Western blotting analysis.Half an hour before 100μg/L rMIF challenged,Y27632 was added to the two groups.After challenged for 48h,total RNA and protein were extracted.Expression of collagen Ⅰ mRNA and protein were analyzed by means of RT-PCR and Western blotting.Results：After 100μg/L rMIF stimulation for 0h,6h,12h,24h or 48h,the concentration of TGF-β1 protein in culture supernants was observed no difference comparing with those of control（P0.05）.After 100μg/L rMIF stimulation for 48h,the expression of collagen Ⅰ mRNA and protein increased in MRC-5 cells（All P0.01）.Y27632 pretreatment prevented the increase（All P0.01）.Conclusion：rMIF increased the collagen Ⅰ synthesis of MRC-5 cell and Rho-kinase regulates MIF-induced the synthesis.These results suggest that MIF may play important roles in the pathogenesis of airway remodeling.

关 键 词：巨噬细胞移动抑制因子 人胚肺成纤维细胞 胶原 RHO 

分 类 号：R562.25[医药卫生—呼吸系统] R361.3[医药卫生—内科学]