摘要
目的探讨重组人Elafin对炎症损伤因子中性粒细胞弹性蛋白酶(NE)攻击气道上皮保护作用的分子机制。方法通过构建人中性粒细胞弹性蛋白酶抑制剂Elafin真核表达载体pEGFP-C1-Elafin,并将其转染入NCI-H292细胞中,再用中性粒细胞弹性蛋白酶(NE)刺激24h后,用底物法测定培养上清中NE的活性,Westernbolt检测细胞中ZO-1表达。结果转染Ela-fin+NE组培养上清液中NE活性与转染空载体的细胞相比较明显降低,而细胞内ZO-1蛋白含量明显增高。结论NE是引起气道慢性炎症的终效因子,通过转染重组人Elafin可对抗NE破坏气道上皮完整性的作用,增强气道抗感染能力。
Objective To explore the molecule mechanism of protection effect of recombinant human elafin gene transfection and expression to neutrophil elastase （NE） attacking airway epithelium. Methods Through constructing eukaryotic expression vector pEGFP- C1-Elafin and transfecting into NCI-H292 cells, the cells with NE were then stimulated. After 24 hours, substrate detection technique and western bolt were respectively performed to detect the vital force unit of NE and expression of ZO-1 in cells. Results After stimulation with NE, comparing with transfecting pEGFP-C1 NCI-H292 cells, the vital force unit of NE in transfecting pEGFP-C1-Elafin NCI-H292 cells media obviously decreased, and the expression of ZO-1 in transfecting pEGFP-C1-Elafin NCI-H292 cells obviously increased. Condusion Transfecting recombinant human dafin gene into airway epithdial cells could maintenance airway apithelium integrity and enhance anti-infection capability of airway.
出处
《临床肺科杂志》
2007年第2期113-114,共2页
Journal of Clinical Pulmonary Medicine
