GFP-hDaxx融合蛋白高表达下调活化巨噬细胞分泌TNF-α和IL-1β  

TNF-α and IL-1β secreted from LPS activated activated macrophages down-regulated by over-expression of GFP-hDaxx fusion protein

作　　者：万艳平[1] 谭立志[1] 刘安元[1] 余敏君[1] 占利生[1] 粟盛梅[1] 

机构地区：[1]南华大学微生物学与免疫学教研室,湖南衡阳421001

出　　处：《中华微生物学和免疫学杂志》2003年第12期961-963,共3页Chinese Journal of Microbiology and Immunology

基　　金：湖南省教育厅资助项目 (No.0 2C3 91) ;湖南省卫生厅资助项目 (No .ZD0 2 2 0 )

摘　　要：目的　研究GFP hDaxx融合蛋白瞬时高表达对活化巨噬细胞分泌活性的影响。方法构建GFP hDaxx融合蛋白真核细胞表达载体pEGFP/hDaxx。将其质粒转染巨噬细胞 ,用荧光显微镜观察GFP hDaxx融合蛋白或GFP的表达与定位 ,并利用Westernblot检测表达产物。通过GFP hDaxx融合蛋白在巨噬细胞内的瞬时高表达 ,在LPS诱导激活巨噬细胞后 0 .5h、4h、12h时 ,测定活化巨噬细胞分泌细胞因子TNF α和IL 1β的含量。结果　GFP hDaxx融合蛋白在巨噬细胞内瞬时高表达且定位于细胞核 ,GFP定位于细胞核与细胞浆。GFP hDaxx融合蛋白在巨噬细胞内的高表达 ,使LPS诱导活化的巨噬细胞分泌TNF α和IL 1β量明显降低 (在 4h及 12h时与对照组差异有显著性 ,P <0 .0 0 1)。结论　hDaxx瞬时高表达可下调活化巨噬细胞分泌细胞因子TNF α和IL 1β。Objective To study the effect of over-expression of GFP-hDaxx fusion protein on the content of TNF-α and IL-1β secreted by the LPS activated macrophages. Methods The eukaryotic expression vector of GFP-hDaxx fusion protein was constructed by hDaxx cDNA fragments subcloned to pEGFP. Twenty four hours after transfection of pEGFP/hDaxx into macrophages, the expression of GFP or GFP-hDaxx was examined with fluorescent microscopy. The macrophage culture media were replaced with a medium containing LPS. During further incubation period after changing media, the content of TNF-α and IL-1β in the media were tested quantitatively with EIA kit at 0.5?h, 4?h and 12?h, respectively. Results Transfected into macrophages, the pEGFP/hDaxx over-expressed GFP-hDaxx and GFP that located respectively in nuclei and cytoplasma. And the secretions of TNF-α and IL-1β reduced significantly at 4?h and 12?h. Conclusion The over-expression of hDaxx fusion protein can down-regulate the secretions of TNF-α and IL-1β in the LPS activated macrophages.

关 键 词：GFP-hDaxx融合蛋白 活化巨噬细胞 分泌 TNF-Α IL-1Β 

分 类 号：R392.12[医药卫生—免疫学]