文档介绍：天然抗生素肽elafin与铜绿假单胞菌的相互作用
作者:聂晓红周向东黄长武
【摘要】目的观察天然抗生素肽elafin与铜绿假单胞菌的相互作用。方法体外细胞培养气道上皮A549细胞,将已构建好elafin克隆载体通过脂质体转染系统转染到A549细胞中,空质粒载体作为其对照,倒置荧光显微镜观察转染效果。空载体转染A549细胞(正常组)以及转染后的A549细胞分别用细胞培养液(对照组)和铜绿假单胞菌上清液(实验组)继续孵育24h。用ELISA法检测每组细胞上清液中elafin水平,Western Blot法检测细胞内elafin含量;孵育24h后,每组上清液取1ml加入到铜绿假单胞菌菌悬液中培养24h,用倍比稀释法测细菌数量。结果对照组细胞的上清液中elafin水平(±)和细胞内elafin蛋白含量(±),与正常组[(±)和(±)]有显著差异(均为P<)。上述指标实验组[(±)和(±)]与对照组相比明显增加(均为P<);对照组细胞上清液中细菌数量[(±)×106/ml]与正常组细胞上清液中的[(±)×106/ml]也有差异(P<),实验组[(±)×106/ml]与对照组相比细菌数明显减少(P<)。细胞上清液中elafin水平与细菌数呈一定的负相关(r=-,P<)。结论铜绿假单胞菌能诱导elafin产生,反之elafin亦有一定的杀铜绿假单胞菌作用。
【关键词】 Elafin; 铜绿假单胞菌; 气道上皮细胞
ABSTRACT Objective To observe the interaction of the natural antibiotic elafin and in airway epithelial cells. Methods After cultivating A549 cells in vitro, the constructed pEGFP-N1-elafin eukaryotic expression vector was transfected into the cells by M 2000, and empty plasmid as a vector control. The A549 cells with pEGFP-N1-elafin vector were incubated for 24 hours in DMEM medium (control group), and the supernatant of (experimental group), and the cells with empty plasmid vector as normal group. The level of elafin secretion in supernatants and the content of elafin in the cells were detected by ELISA and Western Blot respectively. M