siRNA抑制IRE1α蛋白表达对巨噬细胞凋亡的影响

摘要

Objective This study is in order to explore the effect of FC-induced macrophage apoptosis by inhibition IRE1αprotein expression, After accumulation of free cholesterol (FC) in the ER. Methods Wild-type and IRE1α-deifcient peritoneal macrophages from C57BL6/J mice were incubated for 8 h with medium alone, or medium containing acetyl-LDL and compound 58035 (FC-loaded conditions). The apoptosis percentages of macrophages were detected with lfow cytometry (FCM) after the staining of Annexin-V and PI. Results After 8 h of incubated with medium containing AcLDL and compound 58035 (FC-loaded conditions) in our cell culture model, about (21.83±2.47)%of the cells have been detected. But cell apoptosis was markedly reduced in FC loading IRE1α-deifcient macrophages compared with wild-type macrophages. Conclusions It can be conclusion that excess intracellular FC accumulation is an important reason for macrophage apoptosis, and IRE1αis an integral membrane protein of the ER that is a key signaling step in cholesterol-induced apoptosis in macrophages, activated by stress in the ER.%　　目的研究游离胆固醇诱导内质网应激状态下，抑制IRE1α蛋白表达后对巨噬细胞凋亡的影响。方法取自C57BL6/J小鼠腹腔的野生型巨噬细胞和用IRE1α特异性siRNA干扰的巨噬细胞分别在普通培养基及普通培养基加胆固醇乙酰转移酶抑制剂和乙酰低密度脂蛋白（促进游离胆固醇聚集条件下）中进行培养，分组孵育8h后Annexin-V和碘化丙锭双染色后流式细胞仪检测细胞凋亡。结果在FC-loaded状态下经8h孵育，野生型巨噬细胞组大量巨噬细胞凋亡，达到（21.83±2.47）%；siRNA干扰组巨噬细胞的凋亡明显减少。