Maresin 1促进IL-4诱导小鼠巨噬细胞极化的研究

Maresin 1 Promotes the Interleukin-4-induced Polarization of Macrophage in Murine

目的研究maresin 1(MaR1)对白细胞介素4(interleukin 4,IL-4)诱导的RAW264.7巨噬细胞极化的影响以及其相关信号通路。方法将不同浓度的MaR1与IL-4刺激的RAW264.7小鼠巨噬细胞进行孵育,Western blot检测巨噬细胞M2型标志物精氨酸酶1(arginase 1,Arg1)和几丁质酶-3样蛋白3(chitinase 3-like 3,Ym1)的表达水平。将RAW264.7巨噬细胞分为两组进行干预:IL-4组和MaR1+IL-4组;在不同时间点,采用Western blot和免疫荧光检测胞浆磷酸化信号转导及转录激活因子6(signal transducer and activators of transcription 6 phosphorylation,p-STAT6)以及胞核过氧化物酶增殖活化受体γ(peroxisome proliferator activated receptor-gamma,PPARγ)的表达水平。将RAW264.7巨噬细胞加入信号转导及转录激活因子6(signal transducer and activators of transcription6,STAT6)拮抗剂Leflunomide或PPARγ特异拮抗剂GW9662来检测细胞M2型标志物(Arg1和Ym1)的表达水平。结果和其他组比较,10 nmol/L MaR1刺激IL-4诱导的巨噬细胞M2标志物Arg1和Ym1表达增多,组间比较具有统计学差异(Arg1:F=10.96,P<0.05;Ym1:F=10.55,P<0.05),此外,与IL-4相比,MaR1促进IL-4诱导的STAT6磷酸化和PPARγ的核转位时间提前,表达上调(F=17.62、23.45,P均<0.001)。而阻断STAT6的激活以及PPARγ的表达可以降低MaR1诱导的M2型标志物Arg1和Ym1的表达,组间比较差异具有统计学意义(Arg1:F=6.698,P<0.05;Ym1:F=5.769,P<0.05)。结论 MaR1可以刺激RAW264.7巨噬细胞向IL-4诱导的M2型巨噬细胞极化,其作用是通过激活STAT6和PPARγ信号通路实现。

Objective To figure out whether maresin1（MaR1）possess interleukin4（IL-4）-induced alternative activation of RAW264.7 macrophages in murine,and analyze its mechanism and pathways.Methods RAW264.7 murine macrophages were stimulated and incubated with different concentrations of MaR1 after stimulation with IL-4.The expression levels of arginase 1（Arg1）and chitinase 3-like 3（Ym1）of macrophage M2-type markers were detected by Western blot.RAW264.7 macrophage cells were divided into two groups：IL-4 group and MaR1＋IL-4 group.The expression levels of signal transducer and activators of transcription 6 phosphorylation（p-STAT6）and peroxisome proliferator activated receptor-gamma（PPARγ）were detected by Western blot and immunofluorescence at different time points.And the expression levels of macrophage M2-type markers（Arg1 and Ym1）were detected after adding STAT6 antagonist Leflunomide or PPARγspecific antagonist GW9662.Results MaR1（10 nmol/L）significantly increased the expression of M2 macrophage phenotype markers,such as Arg1 and Ym1 in IL-4-induced RAW264.7 macrophages（Arg1：F=10.96,P〈0.05;Ym1：F=10.55,P〈0.05）,and which could enhance the expressions of STAT6 phosphorylation and bring forward the times of PPARγnuclear translocation（F=17.62,23.45,all P 0.001）.The expressions of Arg1 and Ym1 could be reduced by administering Leflunomide（a STAT6 inhibitor）or GW9662（a PPARγantagonist）（Arg1：F=6.698,P〈0.05;Ym1：F=5.769,P〈0.05）.Conclusion MaR1 promotes IL-4-induced M2 type polarization in murine RAW264.7 macrophages via activating STAT6 and PPARγ signaling pathways.