血红素加氧酶-1抑制香烟烟雾提取物致体外人肺A549细胞黏液高分泌  

Heme oxygenase-1 downregulates mucous hypersecretion induced by cigarette smoke in cultured A549 cells

作　　者：雷蕊霞[1] 周向东[1] 

机构地区：[1]重庆医科大学附属第二医院呼吸内科,重庆400010

出　　处：《基础医学与临床》2010年第3期246-251,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(30770951)

摘　　要：目的探讨血红素加氧酶-1(HO-1)对香烟烟雾提取物(CSE)所致人气道黏液高分泌的影响。方法用CSE刺激A549细胞,复制黏液高分泌细胞模型。分为对照组、CSE组、氯化高铁血红素(Hemin)组和锌原卟啉(ZnPPIX)组,观察黏蛋白(MUC)5AC、表皮生长因子受体(EGFR)、磷酸化(p)-EGFR、HO-1及双功能氧化酶1(Duox1)的表达。用MTT法测定细胞活性;RT-PCR法检测HO-1、Duox1、EGFR及MUC5AC的mRNA;Western blot法检测p-EGFR、EGFR、Duox1和HO-1蛋白;ELISA法检测细胞裂解液中MUC5AC蛋白表达。结果CSE组MUC5AC的mRNA吸光度积分相对值和蛋白水平分别为0.660±0.044和(157±3)μg/mg,较对照组0.412±0.043和(105±8)μg/mg明显升高(P<0.05),p-EGFR蛋白水平、EGFR、HO-1及Duox1的mRNA和蛋白水平也较对照组明显增高。与CSE组相比,Hemin组HO-1mRNA及蛋白进一步增高,而p-EGFR蛋白水平、Duox1、EGFR及MUC5AC的mRNA和蛋白水平明显回降。与对照组相比,ZnPPIX组HO-1mRNA及蛋白水平无明显增高,而p-EGFR蛋白水平、Duox1、EGFR及MUC5AC的mRNA和蛋白水平明显增高。结论HO-1可通过下调Duox1的表达,阻断香烟烟雾诱导EGFR活化的上游信号途径,减少EGFR活化,从而抑制MUC5AC的表达。Objective To investigate the effects of heme oxygenase-1（HO-1） on the signal transduction pathway of mucin（MUC） expression induced by cigarette smoke.Methods The cell model of mucous hypersecretion was made by human lung A549 cell stimulated by cigarette smoke extract（CSE）,the cells were divided into 4 groups：negative control group,CSE treatment group,hemin pre-treatment group and ZnPPIX pre-treatment group.The expression of MUC5AC,epidermal growth factor receptor（EGFR）,p-EGFR,HO-1 and dual oxidase 1（Duox1） were detected.The cell activity was assessed by methyl thiazolyl tetrazolium method.The change of HO-1 mRNA,Duox1 mRNA,EGFR mRNA and MUC5AC mRNA was examined by reverse transcriptase-polymerase chain reaction.The protein expression of EGFR,p-EGFR,Duox1 and HO-1 was measured by Western blot,while the protein expression changes of MUC5AC were detected by ELISA.Results The expression level of MUC5AC mRNAand its protein in the CSE group increased significantly （P0.05） as compared to those in the control group[0.412±0.043,（105±8） μg/mg.The mRNA and protein of EGFR,Duox1 and HO-1,the protein of p-EGFR increased significantly as compared to the control group.After the cells being pre-treated with hemin,the mRNA and protein of HO-1 increased significantly,while the mRNA and protein of Duox1,EGFR and MUC5AC,the protein of p-EGFR decreased significantly as compared to the CSE group.After the cells were pre-treated with ZnPPIX,the increase of HO-1 mRNA and protein was not significant as compared to the control group,while the mRNA and protein of Duox1,EGFR and MUC5AC,the protein of p-EGFR increased significantly.ConclusionHO-1 decreased the level of Duox1,blocked ligand-dependent EGF-R activation and decreased the expression levels of MUC5AC.

关 键 词：黏蛋白类 香烟烟雾 受体 表皮生长因子 血红素加氧酶-1 

分 类 号：R363[医药卫生—病理学]