摘要
利用真核表达载体pEGFP/hDaxx在巨噬细胞中高表达hDaxx ,研究hDaxx对巨噬细胞凋亡的影响。将pEGFP/hDaxx转染巨噬细胞 ,Westernblot鉴定Daxx的表达 ,荧光显微镜观察GFP hDaxx融合蛋白在细胞内的表达及定位 ,用地塞米松诱导巨噬细胞凋亡 ,细胞用Giemsa染色后 ,观察细胞形态变化并计算巨噬细胞凋亡率。结果显示 :GFP hDaxx融合蛋白在巨噬细胞中成功表达并定位于细胞核 ,且GFP hDaxx融合蛋白不影响Daxx的特异性 ;地塞米松对照组与pEGFP对照组凋亡率无显著性差异 (P >0 0 5 ) ,而pEGFP/hDaxx组凋亡率显著降低 (P <0 0 1) ,即hDaxx即时高表达降低地塞米松诱导的巨噬细胞凋亡率。Daxx为调控蛋白 ,GFP hDaxx融合蛋白在巨噬细胞中表达并定位于细胞核 ,且hDaxx即时高表达可抑制地塞米松诱导的巨噬细胞凋亡。
In order to study the effect of over-expression of hDaxx on the dexamethasone-induced apoptosis in macrophages, macrophages were transfected with eukaryotic expression vectors pEGFP/hDaxx and pEGFP respecrtively, and the expression of Daxx was identified by Western blotting. In addition, the expression and localization of GFP-hDaxx fusion protein in macrophages were observed under fluorescent microscopy. Then, dexamethasone was added to macrophages to induce apoptosis, and the changes on cellular morphology were observed by Giemsa staining, and the apoptotic rate of macrophages was there by calculated. It was found that GFP-hDaxx fusion protein was successfully expressed on macrophages and localized in nuclei of macrophages. The presence of GFP-hDaxx fusion protein did not interfere with the specificity of Daxx. Besides these, there was a remarkable difference between pEGFP/hDaxx group and the dexamethasone control group and pEGFP control group, all of these indicated that transient over-expression of hDaxx could decrease the apoptotic rate of macrophages induced by dexamethasone.
出处
《现代免疫学》
CAS
CSCD
北大核心
2004年第4期276-278,共3页
Current Immunology
基金
湖南省教育厅 2 0 0 2年课题资助项目 ( 0 2C3 91)
