免疫补体调节蛋白C1抑制物对巨噬细胞极化的作用

Complement regulation protein C1 inhibitor regulates macrophage polarization

目的:研究免疫补体调节蛋白C1抑制物(C1INH)调节巨噬细胞向经典活化巨噬细胞(M1)和选择性激活巨噬细胞(M2)的极化作用。方法:首先从人血中分离单核细胞后,单核细胞在巨噬细胞集落刺激因子(M-CSF)或粒细胞-巨噬细胞集落刺激因子(GM-CSF)作用下转变为静态巨噬细胞;静态巨噬细胞在干扰素-γ(IFN-γ)或白细胞介素4(IL-4)+IL-13刺激作用下分别转化为M1或M2。通过流式细胞仪观察C1INH对巨噬细胞CD14、CD163、CD206表型标志物的作用;应用RTPCR分析C1INH对巨噬细胞细胞因子、趋化因子、相关酶基因表达影响;利用Western blot方法,探讨在炎症条件下C1INH对巨噬细胞CD14结合Toll样受体4(TLR4)的影响。结果:C1INH抑制M-CSF源性M1的CD14、CD163和GM-CSF源性M2的CD206表型。C1INH减少M1的肿瘤坏死因子(TNF-α)和IL-6表达,而增加M2的15脂氧合酶(ALOX15)和IL-10表达。C1INH抑制M1的诱导型一氧化氮合酶(i NOS) mRNA,而提高M2的精氨酸酶1(Arg1) mRNA表达。C1INH促进M1的杀菌活性和M2对菌的吞噬功能。C1INH阻断CD14-TLR4介导信号传导通路。结论:C1INH调节巨噬细胞极化。

Objective:To study regulation of complement regulation protein C1inhibitor(C1INH)in macrophage polarization.Methods:After the isolated human monocytes were incubated with M-CSF or GM-CSF,these cells were induced with IFN-γfor M1or IL-4+IL-13for M2.CD14,CD163and CD206on the surface of M1and M2were detected by analysis of flow cytometry.The mRNA levels of chemokines and cytokines were analyzed by RT-PCR.CD14binding to Toll-like receptor4(TLR4)were examined by Western blot.Results:C1INH regulated expression of CD14,CD163and CD206on M1and M2.C1INH decreased releases of TNF-α,IL-6and iNOS in M1and increased levels of ALOX15,IL-10and Arg1in M2.Administration of C1INH improved bactericidal activity in M1and phagocytosis function M2.Inhibition of C1INH in CD14binding activity interfered in TLR4-mediated signaling pathway.Conclusion:C1INH regulates macrophage polarization.