阿魏酸抑制p38 MAPK信号传导对巨噬细胞M1极化的作用

Effect of ferulic acid on M1 polarization of macrophages by inhibiting p38 MAPK signaling

目的研究阿魏酸对脂多糖(LPS)与干扰素-γ(IFN-γ)诱导巨噬细胞M1极化的影响与作用机制。方法THP-1细胞加入佛波酯(PMA)诱导成M0型巨噬细胞,使用LPS+IFN-γ诱导成M1型巨噬细胞。采用CCK-8法确定阿魏酸对M1型巨噬细胞的实验浓度范围;流式细胞术检测阿魏酸对M1型巨噬细胞极化情况;酶联免疫吸附实验(ELISA)和实时荧光定量PCR法检测阿魏酸对M1型巨噬细胞分泌产物肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)mRNA和蛋白表达的影响;Western blot法检测p38 MAPK、p38 MAPK磷酸化(p-p38 MAPK)蛋白表达水平的变化。结果阿魏酸浓度在80μmol/L以下对M1型巨噬细胞的活性无影响。在阿魏酸作用下,与对照组比较,M1型巨噬细胞标志物CD80+的表达显著降低(P<0.05);同时,阿魏酸能抑制M1型巨噬细胞分泌的TNF-α、IL-1β和p38 MAPK磷酸化(p-p38 MAPK)蛋白表达,其作用呈剂量依赖性。结论阿魏酸抑制M0型巨噬细胞向M1型极化,降低促炎因子表达,其抗炎作用机制与抑制p38 MAPK信号通路活化有关。

Objective To study the effect and mechanism of ferulic acid on M1 polarization of macrophages induced by LPS and IFN-γ.Methods THP-1 cells were induced into M0 type macrophages by adding PMA,and M1 type macrophages were induced using LPS+IFN-γ.The concentration range of ferulic acid on M1 macrophages was determined by CCK-8 method.Flow cytometry was used to detect the polarization of M1 macrophages induced by ferulic acid.ELISA and real-time quantitative PCR were used to detect the effects of ferulic acid on the expression of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)mRNA and protein by M1 type macrophages.Western blot was used to detect the expression of p38 MAPK and p-p38 MAPK.Results Ferulic acid concentration below 80μmol/L had no effect on the activity of M1 type macrophages.Compared with the control group,the expression of M1 type macrophage marker CD80+was significantly decreased under ferulic acid treatment(P<0.05).Ferulic acid also inhibited the expression of TNF-α,IL-1βand p-p38 MAPK by M1 type macrophages in a dose-dependent manner.Conclusion Ferulic acid can inhibit the polarization of M0 macrophages towards M1 type and reduce the expression of pro-inflammatory factors,which is related to the inhibition of p38 MAPK signaling pathway.