即早基因c-fos在THP-1单核巨噬细胞亚型极化中的差异表达

Differential expression of immediately early gene c-fos in THP-1 macrophage subtype polarization

目的:探讨即早基因c-fos在THP-1巨噬细胞亚型极化过程中的表达变化。方法:运用PMA刺激诱导THP-1单核细胞极化为巨噬细胞,观察c-fos在单核细胞极化过程中的表达变化;在PMA刺激的基础上,分别运用LPS和IL-4诱导THP-1巨噬细胞向M1及M2亚型极化,实时定量PCR及Western blot技术分析刺激24 h时,细胞亚型标记物CD274、CD86和CD163的表达变化,并动态观察诱导极化过程中,c-fos的表达情况。结果:c-fos在PMA刺激THP-1单核细胞分化为巨噬细胞过程中蛋白和mRNA水平显示上调;LPS诱导THP-1巨噬细胞极化为M1型过程中,c-fos蛋白和mRNA水平表达降低,其特异性标记物在24 h呈现出M1型极化的特点(CD86蛋白表达升高,CD274、CD163蛋白表达降低);IL-4诱导THP-1巨噬细胞极化为M2型过程中,c-fos蛋白和mRNA水平表达升高,其特异性标记物在24 h表现出M2型极化的特点(CD86蛋白表达降低,CD274、CD163蛋白表达升高)。结论:c-fos参与了THP-1单核细胞向巨噬细胞极化的过程,并且可能通过抑制巨噬细胞M1亚型形成,促进巨噬细胞向M2亚型极化的作用参与巨噬细胞的亚型极化及其功能调节中。

Objective:To investigate the expression of immediate early gene c-fos in THP-1 macrophage subtype polarization.Methods:PMA was used to induce the polarization of THP-1 monocytes to macrophages,and the expression of c-fos in the polarization process was observed.After PMA treatment,LPS or IL-4 were used alone to induce the polarization of THP-1 macrophages to the M1 or M2 subtypes.Subsequently,real-time quantitative PCR and western-blot were used to analyze the changes in the expressions of the cell subtype markers CD274,CD86 and CD163.Meanwhile,the expression of c-fos in the polarization process was observed dynamically.Results:The levels of c-fos protein and mRNA expressions were up-regulated during PMA-induced polarization of THP-1 monocytes.The protein and mRNA expressions of c-fos were significantly decreased during the polarization of THP-1 cells into M1 macrophages induced by LPS.The specific markers showed the characteristics of M1 macrophages polarization at 24 h(CD86 protein increased,CD274 and CD163 protein decreased).The protein and mRNA expressions of c-fos were significantly increased during the polarization of THP-1 cells into M2 macrophages induced by IL-4.The specific markers showed the characteristics of M2 macrophages polarization at 24 h(CD86 protein decreased,CD274 and CD163 protein increased).Conclusion:C-fos plays an important role in the polarization of THP-1 monocytes to macrophages.Moreover,it may be involved in the regulation of macrophage subtype polarization,by inhibiting the formation of M1 macrophage and promoting the polarization of macrophages to the M2 subtype.