Aire对巨噬细胞极化影响的研究

Effects of Aire on macrophage polarization

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RAW264.7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-1α、i NOS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0.5μg/ml浓度时,RAW264.7细胞中M1型巨噬细胞产物IL-1α、i NOS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-1α和i NOS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。

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-1α,i NOS and IL-6）,M2 a related molecules（ Arg-1） and M2 b related molecule（ IL-10） were detected by Real-time PCR. Results： The expression of IL-1α,IL-6 and inducible nitric oxide synthase（ i NOS）,the products of M1 macrophages,were significantly upregulated in RAW264. 7 cells treated with LPS at 0. 5 μg/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 i NOS 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 M2 a and M2 b macrophage polarization.