Azithromycin Polarizes Macrophages to an M2 Phenotype via Inhibition of the STAT1 and NF-κB Signaling Pathways
Azithromycin Polarizes Macrophages to an M2 Phenotype via Inhibition of the STAT1 and NF-κB Signaling Pathways
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DOI：
10.4049/jimmunol.1801228
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发表时间：
2019-08-15
影响因子：
4.4
通讯作者：
Feola, David J.
中科院分区：
文献类型：
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作者：
Haydar, Dalia;Cory, Theodore J.;Feola, David J.
阿奇霉素可有效控制囊性纤维化患者的过度炎症和减缓肺功能的长期下降。我们以前证明，该药物将巨噬细胞极化向另一种抗炎表型转移。在这项研究中，我们通过改变NF-κ B和STAT 1通路来研究阿奇霉素的免疫调节机制。将J774鼠巨噬细胞铺板、极化（用IFN-γ、IL-4/-13或用阿奇霉素加IFN-γ）并用LPS刺激。通过Western blot、均相时间分辨荧光法、核转位法和免疫荧光法评估阿奇霉素对NF-κ B和STAT 1信号介质的影响。评价药物对巨噬细胞酶基因和蛋白表达的影响，作为替代性巨噬细胞活化的标志物。阿奇霉素通过减少p65核转位阻断NF-κ B活化，尽管I κ B α降解的钝化至少部分是由于IKK β激酶活性的降低。阿奇霉素浓度增加与IKK β蛋白表达增加之间存在直接相关性。此外，与IKK β抑制剂IKK 16孵育降低了阿奇霉素处理的细胞中的激酶表达和活性，但在用IL-4和IL-13处理的细胞中没有。重要的是，阿奇霉素治疗还以浓度依赖性方式降低了STAT 1磷酸化，这种作用在IKK 16治疗中逆转。我们的结论是，阿奇霉素抗炎机制涉及通过药物对p65核转位和IKK β的影响抑制STAT 1和NF-κ B信号通路。
Azithromycin is effective at controlling exaggerated inflammation and slowing the long-term decline of lung function in patients with cystic fibrosis. We previously demonstrated that the drug shifts macrophage polarization toward an alternative, anti-inflammatory phenotype. In this study we investigated the immunomodulatory mechanism of azithromycin through its alteration of signaling via the NF-kappa B and STAT1 pathways. J774 murine macrophages were plated, polarized (with IFN-gamma, IL-4/-13, or with azithromycin plus IFN-gamma) and stimulated with LPS. The effect of azithromycin on NF-kappa B and STAT1 signaling mediators was assessed by Western blot, homogeneous time-resolved fluorescence assay, nuclear translocation assay, and immunofluorescence. The drug's effect on gene and protein expression of arginase was evaluated as a marker of alternative macrophage activation. Azithromycin blocked NF-kappa B activation by decreasing p65 nuclear translocation, although blunting the degradation of I kappa B alpha was due, at least in part, to a decrease in IKK beta kinase activity. A direct correlation was observed between increasing azithromycin concentrations and increased IKK beta protein expression. Moreover, incubation with the IKK beta inhibitor IKK16 decreased arginase expression and activity in azithromycin-treated cells but not in cells treated with IL-4 and IL-13. Importantly, azithromycin treatment also decreased STAT1 phosphorylation in a concentration-dependent manner, an effect that was reversed with IKK16 treatment. We conclude that azithromycin anti-inflammatory mechanisms involve inhibition of the STAT1 and NF-kappa B signaling pathways through the drug's effect on p65 nuclear translocation and IKK beta.