糖皮质激素诱导的亮氨酸拉链蛋白(GILZ)在人气道上皮细胞中的表达及对MAPK信号通路的影响  

The expression of glucocorticoid-induced leucine zipper(GILZ) and its effect to MAPK signaling pathway in human airway epithelial cells

作　　者：刘静月[1] 符州[2] 张明香[2] 刘恩梅[2] 罗征秀[2] 王莉佳[2] 

机构地区：[1]重庆医科大学附属儿童医院发育疾病研究教育部重点实验室,重庆400014 [2]重庆医科大学附属儿童医院呼吸中心,重庆400014

出　　处：《中国免疫学杂志》2012年第8期737-740,751,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金项目(81070014);重庆市卫生局重点项目(2010-01-46)资助

摘　　要：目的:探讨糖皮质激素诱导的亮氨酸拉链蛋白(GILZ)在人气道上皮细胞9HTE0中的表达以及对MAPK信号通路因子Raf-1、Mek1/2、Erk1/2的影响。方法:采用RT-PCR及Western blot法检测地塞米松(Dex)作用后GILZ mRNA及蛋白的表达,同时细胞免疫荧光法检测GILZ蛋白的定位;合成三条GILZ-SiRNA分别转染人气道上皮细胞9HTE0,用Q-PCR及Western blot法筛选出沉默效果最佳的一条;Western blot法检测MAPK信号通路因子Raf-1、Mek1/2、Erk1/2磷酸化蛋白及其总蛋白的表达。结果:Dex能够明显刺激人气道上皮细胞9HTE0GILZ mRNA及蛋白的表达,GILZ蛋白主要定位在细胞浆中,Dex抑制了Raf-1、Mek1/2、Erk1/2磷酸化蛋白的表达,而GILZ-SiRNA转染后,Raf-1.Mek1/2、Erk1/2磷酸化水平有所回升。结论:糖皮质激素作为支气管哮喘一线用药,虽然能够诱导GILZ表达并发挥一定的抗炎作用,但同时GILZ却抑制了在气道上皮修复中起重要作用的MAPK信号通路的激活,这为临床上研究哮喘防治新措施提供了理论依据。Objective：To explore the expression of glucocorticoid-induced leucine zipper and the effect of GILZ to Raf-1,Mek1/2,Erk1/2 of MAPK signaling pathway in human airway epithelial 9HTE0 cells.Methods：RT-PCR was used to detect GILZ mRNA.GILZ protein was tested by Western blot and located by immunofluorescence.Three synthetic GILZ-SiRNAs were respectively transfected into 9HTE0 cells to screen out the best GILZ-SiRNA.Western blot assayed the phosphorylated and total proteins of Raf-1,Mek1/2,Erk1/2.Results：GILZ mRNA and protein increased obviously with Dexamethasone stimulation and GILZ protein mainly expressed in cell cytoplasm of 9HTE0 cells.The phosphorylation of Raf-1,Mek1/2,Erk1/2 protein of which Dexamethasone inhibited the expression rebounded after GILZ-SiRNA was transfected.Conclusion：Glucocorticoid as the fist line treatment of asthma induced the expression of GILZ that played a certain anti-inflammatory effect,but GILZ inhibited the activation of MAPK signaling pathway which promoted the repair process of airway epithelial cells.This provided a theoretical basis for new therapeuticmeasures of clinical research in asthma.

关 键 词：糖皮质激素 GILZ 气道上皮细胞 MAPK 支气管哮喘 

分 类 号：R725.6[医药卫生—儿科]