【摘要】 目的　探讨影响Ⅰ型IL 1受体 (IL 1RⅠ )转位至焦点附着斑的相关因素。方法　将含有绿色荧光蛋白 (GFP)基因的重组型和突变型 (C末端突变 )IL 1RⅠ质粒转染进入成纤维细胞 ,细胞生长于有或无纤维粘连蛋白的培养皿上 ,用激光共聚焦显微镜直接观察所表达的融合蛋白在细胞内的定位 ,IL 1β刺激后发生转位 ,并观察经免疫细胞化学染色后 ,细胞骨架蛋白Vinculin的位置。结果　转染了重组型IL 1R1质粒的细胞生长在纤维粘连蛋白上 ,表达的受体融合蛋白与Vinculin位于细胞膜的焦点附着斑 ,用IL 1β刺激可致受体转位至细胞内。转染突变型质粒的细胞 ,表达的融合蛋白位于胞质的核周区域 ,IL 1β刺激后不产生转位。 结论　Ⅰ型IL 1受体转位至焦点附着斑需要Vinculin等细胞外基质的参与并与受体C 末端保守结构域有关更多还原

【Abstract】 Objective To study the mechanisms of regulation of translocation of IL 1 receptor to focal adhesions.Methods Fibroblasts transfected with wild type and mutant IL 1 receptor cDNA containing GFP sequence were plated on fibronectin or bare tissue culture plastic. The location of expressing fusion protein at single cell level , translocation after binding with IL 1,co localization of receptor and transmembrane linkage protein vinculin by immunocytochemical staining were determined under confocal microscopy. Results In fibronectin attached cells, the wild type IL 1 receptor fusion protein and vinculin were co located at focal adhesions in cell membrane. Further, the fusion proteins moved into cytoplasm and finally concentrated in nucleus after IL 1 stimulation. In contrast, mutant and extracellular domain IL 1 receptors located in the perinuclear cytoplasm were unaffected by IL 1 stimulation.Conclusions Translocation of the IL 1 receptor to focal adhesions is dependant on conserved domains in the C terminal end of the protein and the presence of cell matrix.更多还原