基础医学与临床 ›› 2007, Vol. 27 ›› Issue (3): 302-305.

• 研究论文 • 上一篇    下一篇

人ELAFIN基因重组转染对气道上皮完整性的维护作用

徐晓艳 邵延春 周向东   

  1. 重庆医科大学第二附属医院呼吸内科 山东省诸城市人民医院综合内科 重庆医科大学第二附属医院呼吸内科
  • 收稿日期:2006-02-28 修回日期:2006-08-04 出版日期:2007-03-25 发布日期:2007-03-25
  • 通讯作者: 徐晓艳

Protective effects of human ELAFINgene recombinant plasmid transfection to airway epithelium integrity

  

  • Received:2006-02-28 Revised:2006-08-04 Online:2007-03-25 Published:2007-03-25

摘要: 目的 探讨转染重组人白细胞弹性蛋白酶抑制剂Elafin基因保护气道上皮的分子机制。方法 构建人Elafin重组质粒pEGFP-C1-Elafin,并转染入人气道黏膜上皮细胞系NCI-H292细胞,将其与多形核粒细胞(PMN)共孵育,经脂多糖(LPS)刺激24h后,用Western blot 检测细胞中紧密连接相关蛋白ZO-1的表达及MTT法测定细胞与胶原的附着能力。结果 经LPS刺激后,Western blot及MTT法发现,与对照组和转染重组质粒组相比,转染空载体的NCI-H292细胞中ZO-1的表达及细胞与胶原附着能力均明显降低(P<0.01);而与对照组相比,转染重组质粒的细胞中ZO-1的表达及细胞与胶原附着能力均未见明显下降。结论 通过转染重组人Elafin可保护气道上皮,增强气道抵抗炎性损伤能力。

Abstract: Objective To explore the molecule mechanism of protective effects of transfecting human elafin gene recombinant plasmid to NCI-H292 .Methods Through constructing eukaryotic expression vector pEGFP-C1-elafin and transfecting it into NCI-H292 cells ,then co-incubation with polymorphonuclear granulocyte(PMN) and stimulating these cells with lipopolysaccharide(LPS).Afater 24hour,Western bolt and MTT technique were respectively performed to detect expression of ZO-1 in cells and adhesion capability of cells with collagen.Results After stimulation with LPS,Western blot experiment showed that the 220-kD zonula occludens-1(ZO-1) protein in transfecting pEGFP-C1 NCI-H292 cells extracts were obviously decreased,but were still present in transfecting pEGFP-C1-elafin NCI-H292 cells extracts.And MTT technique showed that compared with transfecting pEGFP-C1-elafin cells,adhesion capability of transfecting pEGFP-C1 cells with collagen were obviously decreased.Conclusions Transfection of recombinant human elafin gene into airway epithelial cells could maintain airway epithelium and enhance the capability of airway to inflammatory injury.