摘要
目的探讨转染重组人白细胞弹性蛋白酶抑制剂内生多肽(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24 h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。
Objective To explore the molecule mechanism of protective effects of transfecting human elafin gene recombinant plasmid to NCI-H292. Methods Constructe eukaryotic expression vector pEGFP-Cl-elafin and transfectied it into NCI-H292 cells, then co-incubated with polymorphonuclear granulocyte （PMN） with stimulation by lipopolysaccharide （LPS）. Mater 24 hour, 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,Westem blot showed that the 220 ku onula occludens-1 （ ZO-1 ） protein in transfecting pEGFP-Cl NCI-H292 cells extracts were obviously decreased, in transfecting pEGFP-Cl-elafin NCI-H292 cells extracts. And MTT technique showed that compared with transfecting pEGFP-Cl-elafin cells,adhesion capability of transfecting pEGFP-Cl cells with collagen were obviously decreased. Conclusion Transfection of recombinant human elafin gene into airway epithelial cells could maintain airway epithelium and enhance the capability of airway to inflammatory injury.
出处
《基础医学与临床》
CSCD
北大核心
2007年第3期302-305,共4页
Basic and Clinical Medicine
基金
重庆市应用基础研究项目(2003032)
