中文摘要：

目的通过增强型绿色荧光蛋白示踪技术,观察SIGIRR(single Ig IL-1R-related molecule)在人气道上皮细胞株H292中的分布特点及对上皮细胞天然免疫功能的影响。方法构建SIGIRR-EGFP融和蛋白的真核表达载体,运用脂质体转染的方法转染人气道上皮细胞株H292,再利用激光共聚焦显微镜对其进行亚细胞定位研究。经LPS刺激后,ELISA检测转染前后上皮细胞TNF-α分泌水平的差异。结果构建的融和蛋白表达载体SIGIRR-EGFP经EcoRⅠ和BamHⅠ双酶切后,电泳显示其条带大小为4.7 kb和1.23 kb左右,经测序证实SIGIRR的序列与GenBank公布的人cDNA序列完全一致。激光共聚焦显示pEGFP-N1空载体转染表达的绿色荧光蛋白在H292细胞中均匀分布,而重组载体SIGIRR-EGFP表达的融和蛋白在H292细胞核周(膜)和细胞膜边缘上有连续分布现象。经LPS刺激后,pEGFP-N1转染的细胞和未转染细胞TNF-α水平无显著性差异(P>0.05),而SIGIRR-EGFP转染细胞与pEGFP-N1转染细胞相比,显著降低了TNF-α的产生(P<0.01)。结论成功构建...

英文摘要：

Objective To analyze the expression and distribution of SIGIRR-EGFP in human air-way epithelial cells H292 using enhanced green fluorescent protein tracer technology and its impact on immune function of the cells.Methods A eukaryotic expression vector for single Ig IL-1R-related molecule(SIGIRR) fused with enhanced green fluorescent protein(EGFP) gene was constructed and transfected into H292.The subcellular localization of SIGIRR-EGFP was observed by laser confocal microscope.ELISA showed the change of IL-...