脂多糖诱导小鼠巨噬细胞系RAW264.7细胞的活化凋亡作用

目的探讨脂多糖(LPS)诱导小鼠巨噬细胞系RAW264.7细胞活化凋亡的作用。方法体外培养小鼠巨噬细胞系RAW264.7细胞,分别用0.5、1.0、2.5μg/mlLPS刺激RAW264.7细胞24h,一氧化氮(NO)试剂盒检测细胞培养上清中NO水平;用1.0μg/mlLPS分别刺激细胞3d和6d,台盼蓝拒染法检测细胞增殖情况;用1.0μg/mlLPS刺激细胞6d,流式细胞术分析细胞周期及细胞凋亡情况。结果经LPS刺激后24h,RAW264.7细胞培养上清中NO含量明显增加,且具有剂量依赖性;LPS刺激3d和6d后,细胞的增殖均受到抑制,且呈时间依赖性;LPS刺激6d时,细胞周期被阻滞在S期,并出现明显的凋亡。结论LPS具有诱导小鼠巨噬细胞系RAW264.7细胞活化凋亡的作用。

Activation-Induced Apoptosis of Mouse Macrophage RAW264.7 by Lipopolysaccharide

Objective To explore the role of lipopolysaccharide(LPS)in activation-induced apoptosis of mouse macrophage RAW264.7.Methods RAW264.7 cells were cultured in vitro and stimulated with LPS at dosages of 0.5,1.0 and 2.5 μg / ml for 24 h respectively and determined for nitric monooxide(NO)level in culture supernatant by NO kit.The cells were stimulated with 1.0 μg / ml LPS for 3 and 6 d respectively and determined for proliferation level by trypan blue dye exclusion method.The cells were also stimulated with 1.0 μg / ml LPS for 6 d and analyzed for cell cycle and apoptosis by flow cytometry.Results After stimulation with LPS for 24 h,the NO level in culture supernatant of RAW264.7 cells showed a dose-dependent increase.After stimulation with LPS for 3 and 6 d,the proliferation of RAW264.7 cells was inhibited in a time-dependent mode.After stimulation with LPS for 6 d,the cell cycle was blocked at S phase,and obvious apoptosis was observed.Conclusion LPS plays an important role in activation-induced apoptosis of mouse macrophage RAW264.7.