Elafin基因重组气道上皮调节脂多糖诱导的人气道黏液高分泌  

Effects of airway epithelial cells transfected by endogeny polypeptide Elafin on human airway mucus hypersecretion

作　　者：余红梅[1] 李琪[1] 尤列.皮尔曼 维克多.科罗索夫 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 [2]俄罗斯医学科学院远东呼吸生理与病理研究中心,俄罗斯布拉戈维申斯克675000

出　　处：《基础医学与临床》2011年第4期371-376,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(30770951);国家自然科学基金中俄国际合作项目(30910303);中俄政府间合作项目(2009:13-01)

摘　　要：目的探讨天然内生多肽Elafin对脂多糖诱导的气道黏液高分泌的调节机制。方法构建人Elafin重组质粒pEGFP-N1-Elafin,转染正常人支气管上皮细胞HBE16,给予脂多糖(LPS)刺激,激光共聚焦扫描显微镜检测Elafin蛋白含量,Western blot法检测IκBα蛋白含量;荧光素酶报告基因检测系统测定核转录因子-κB(NF-κB)活性;RT-PCR检测Elafin和黏蛋白(MUC)5AC mRNA表达水平;ELISA法分析MUC5AC蛋白相对含量。结果成功构建内生多肽Elafin真核表达载体pEGFP-N1-Elafin并转染HBE16细胞。转染重组Elafin后Elafin蛋白含量及mRNA水平明显增加。与对照组相比,LPS刺激组IκBα蛋白含量降低,NF-κB相对活性明显增强,MUC5AC蛋白含量及mRNA水平也显著增加。转染重组Elafin后再予以LPS刺激,与单纯LPS刺激组相比,IκBα蛋白含量明显增加,NF-κB相对活性显著降低,MUC5AC蛋白含量及mRNA水平也明显降低。结论天然内生多肽Elafin重组气道上皮可下调NF-κB的活性,降低脂多糖诱导的气道黏液高分泌。Objective To explore the effects of transfection of endogeny polypeptide elafin on mucin（MUC）5AC overexpression induced by lipopolysaccharide.Methods Eukaryotic expression vector pEGFP-N1-Elafin was constructed and transfected into HBE16 Cells.Cells were then stimulated by lipopolysaccharide（LPS）.Elafin was detected by immunofluorescence and observed by confocal laser scanning microscopy（CLSM）,the protein level inhibitor of NF-κB（IκB）α was detected by Western blot,the transcription activity of nuclear factor κB（NF-κB） was detected by luciferase reporter gene detection system,MUC5AC protein level was assessed by ELISA,the level of Elafin mRNA and MUC5AC mRNA was detected by RT-PCR,respectively.Results The level of Elafin protein and mRNA was obviously increased after transfection of recombinant Elafin into HBE16 cells,compared with LPS group or control group.There was an significant increase in MUC5AC protein production and mRNA expression,with elevation of NF-κB activity,IκBα protein production was significantly lower than that of control group.Transfected recombinant elafin reduced MUC5AC protein and mRNA level,inhibited NF-κB activity,and increased IκBα protein production,compared with single LPS-stimulated group.Conclusion Transfection of endogeny polypeptide Elafin may inhibit MUC5AC overexpression induced by LPS,and this is through the inhibition by Elafin of NF-κB activiation via effects on IκB pathway.

关 键 词：脂多糖 内生多肽 ELAFIN 黏蛋白 核转录因子-ΚB 

分 类 号：R363[医药卫生—病理学]