Aire 对巨噬细胞极化影响的研究

目的:研究自身免疫调节因子(Aire)对巨噬细胞极化的影响。方法:分别用LPS、IL-4 以及LPS 联合免疫复合物刺激小鼠单核巨噬细胞系RAW264郾7 细胞、稳定表达GFP-Aire 的RAW264.7 细胞(A33-3) 细胞和稳定表达GFP 的RAW264.7 细胞(C1-6),使其向M1(LPS)、M2a(IL-4)和M2b(LPS 联合免疫复合物)型巨噬细胞极化。通过Real-time PCR 检测各组细胞中M1 型巨噬细胞特征分子IL-1、iNOS 和IL-6,M2a 型特征分子Arg-1 和M2b 型特征分子IL-10 的表达水平,研究Aire 对各种类型巨噬细胞极化的影响。结果:LPS 在0.5 g/ ml 浓度时,RAW264.7 细胞中M1 型巨噬细胞产物IL-1 、iNOS和IL-6 基因表达量最高;而IL-4 以及LPS 联合免疫复合物的刺激作用有显著的剂量依赖性,都在浓度最高时RAW264.7 细胞中Arg1(M2a)和IL-10(M2b)基因表达量最高。LPS 刺激后,A33-3 细胞中IL-1 和iNOS 表达水平明显高于C1-6 细胞,IL-6 则相反;IL-4 及LPS 联合免疫复合物刺激后,A33-3 细胞中Arg1 和IL-10 的表达水平明显低于C1-6 细胞。结论:Aire 可能促进巨噬细胞向M1 极化,同时抑制其向M2a 和M2b 极化。

Effects of Aire on macrophage polarization

Objective:To study whether autoimmune regulator (Aire) affects macrophage polarization.Methods: The mouse mononuclear macrophage cell line RAW264.7 cells, stable expressing GFP-Aire protein RAW264.7 cells ( A33-3) and stable expressing GFP protein RAW264.7 cells (C1-6) were stimulated with LPS,IL-4 and LPS combined with immune complex respectively to make the macrophage polarize to M1 (LPS),M2a (IL-4) and M2b (LPS with immune complex).To investigate the effects of Aire on the various types of macrophages polarization,the M1 related molecules (IL-1,iNOS and IL-6),M2a related molecules (Arg-1) and M2b related molecule (IL-10) were detected by Real-time PCR.Results: The expression of IL-1 ,IL-6 and inducible nitric oxide synthase (iNOS),the products of M1 macrophages,were significantly upregulated in RAW264.7 cells treated with LPS at 0.5 g/ml.The expression of Arg-1 and IL-10 mRNA in RAW264.7 cells increased in a dose-dependent manner after stimulation with IL-4 or LPS combined with immune complexes,respectively.M1 macrophage-related marker iNOS and IL-1 increased,whereas IL-6 levels decreased in A33-3 cells compared with C1-6 cells after treatment with LPS.The expression of Arg1 and IL-10 were downregulated in A33-3 cells compared with C1-6 cells after IL-4 and LPS combined with immune complexes stimulation,respectively. Conclusion: Aire may promote M1 macrophage polarization while inhibit M2a and M2b macrophage polarization.