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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。
Objective To investigate the effect of lipopolysaccharide (LPS) on the activation and apoptosis of murine macrophage cell line RAW264.7. Methods RAW264.7 cells were cultured in vitro. RAW264.7 cells were stimulated with 0.5, 1.0 and 2.5 μg / ml LPS for 24 h respectively. Nitric oxide (NO) kit was used to detect NO level in cell culture supernatant. μg / ml LPS for 3 days and 6 days respectively. The proliferation of cells was detected by trypan blue exclusion method. The cells were stimulated with 1.0μg / ml LPS for 6 days. The cell cycle and apoptosis were analyzed by flow cytometry. Results After stimulated with LPS for 24 h, the content of NO in the culture supernatant of RAW264.7 cells increased significantly and in a dose-dependent manner. After LPS stimulation for 3 and 6 days, the proliferation of cells was inhibited in a time-dependent manner. LPS stimulation for 6 days , The cell cycle is blocked in the S phase, and significant apoptosis. Conclusion LPS can induce the apoptosis of mouse macrophage cell line RAW264.7.