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目的:研究γ射线与LPS协同激活小鼠巨噬细胞RAW264.7的效应机制,以及γ射线与LPS诱导钙结合蛋白S100A8的表达及意义。方法:相差显微镜观察细胞形态;流式细胞计数方法检测细胞周期及活性氧介质(ROI)水平;Griess颜色反应测定细胞NO水平;实时定量RT-PCR方法检测细胞S100A8 mRNA水平的表达。结果:γ射线与LPS作用于RAW264.7细胞引起细胞形态改变,部分细胞出现非整倍性,少量细胞出现凋亡,胞内ROI水平、NO水平明显升高,S100A8分子mRNA水平明显升高,而且这些效应几乎都比γ射线或LPS单因素的作用要强。结论:γ射线与LPS协同诱导巨噬细胞激活是细胞周期改变、信使分子水平变化、炎症因子如S100A8的表达等多方面生物效应的综合结果,其中S100A8基因的表达与巨噬细胞功能状态密切相关。
AIM: To investigate the mechanism of γ-ray and LPS synergistically activating RAW264.7 in mouse macrophages and the expression of S100A8, a calcium-binding protein induced by γ-rays and LPS. Methods: Cell morphology was observed by phase contrast microscopy. Cell cycle and reactive oxygen species (ROI) levels were measured by flow cytometry. NO levels were measured by Griess color reaction. The expression of S100A8 mRNA was detected by real-time RT-PCR. Results: γ ray and LPS induced cell morphological changes in RAW264.7 cells, some cells showed aneuploidy, a small amount of cells showed apoptosis, intracellular ROI levels, NO levels were significantly increased, S100A8 mRNA levels were significantly increased, And almost all of these effects are stronger than single-factor gamma rays or LPS. CONCLUSION: The synergistic effect of γ-rays and LPS on the activation of macrophages is a combination of biological effects such as cell cycle changes, changes of the level of messenger molecules and the expression of inflammatory factors such as S100A8. The expression of S100A8 is closely related to the functional status of macrophages .