Dougherty Edward J, Elinoff Jason M, Ferreyra Gabriela A, Hou Angela, Cai Rongman, Sun Junfeng, Blaine Kevin P, Wang Shuibang, Danner Robert L
From the Critical Care Medicine Department, Clinical Center, National Institutes of Health, Bethesda, Maryland 20892
From the Critical Care Medicine Department, Clinical Center, National Institutes of Health, Bethesda, Maryland 20892.
J Biol Chem. 2016 Nov 4;291(45):23628-23644. doi: 10.1074/jbc.M116.732248. Epub 2016 Sep 20.
Glucocorticoids are commonly used to treat inflammatory disorders. The glucocorticoid receptor (GR) can tether to inflammatory transcription factor complexes, such as NFκB and AP-1, and trans-repress the transcription of cytokines, chemokines, and adhesion molecules. In contrast, aldosterone and the mineralocorticoid receptor (MR) primarily promote cardiovascular inflammation by incompletely understood mechanisms. Although MR has been shown to weakly repress NFκB, its role in modulating AP-1 has not been established. Here, the effects of GR and MR on NFκB and AP-1 signaling were directly compared using a variety of ligands, two different AP-1 consensus sequences, GR and MR DNA-binding domain mutants, and siRNA knockdown or overexpression of core AP-1 family members. Both GR and MR repressed an NFκB reporter without influencing p65 or p50 binding to DNA. Likewise, neither GR nor MR affected AP-1 binding, but repression or activation of AP-1 reporters occurred in a ligand-, AP-1 consensus sequence-, and AP-1 family member-specific manner. Notably, aldosterone interactions with both GR and MR demonstrated a potential to activate AP-1. DNA-binding domain mutations that eliminated the ability of GR and MR to cis-activate a hormone response element-driven reporter variably affected the strength and polarity of these responses. Importantly, MR modulation of NFκB and AP-1 signaling was consistent with a trans-mechanism, and AP-1 effects were confirmed for specific gene targets in primary human cells. Steroid nuclear receptor trans-effects on inflammatory signaling are context-dependent and influenced by nuclear receptor conformation, DNA sequence, and the expression of heterologous binding partners. Aldosterone activation of AP-1 may contribute to its proinflammatory effects in the vasculature.
糖皮质激素常用于治疗炎症性疾病。糖皮质激素受体(GR)可与炎症转录因子复合物(如NFκB和AP-1)结合,并反式抑制细胞因子、趋化因子和黏附分子的转录。相比之下,醛固酮和盐皮质激素受体(MR)主要通过尚未完全明确的机制促进心血管炎症。尽管已表明MR可微弱抑制NFκB,但其在调节AP-1中的作用尚未明确。在此,使用多种配体、两种不同的AP-1共有序列、GR和MR的DNA结合结构域突变体以及核心AP-1家族成员的siRNA敲低或过表达,直接比较了GR和MR对NFκB和AP-1信号传导的影响。GR和MR均抑制NFκB报告基因,而不影响p65或p50与DNA的结合。同样,GR和MR均不影响AP-1的结合,但AP-1报告基因的抑制或激活以配体、AP-1共有序列和AP-1家族成员特异性的方式发生。值得注意的是,醛固酮与GR和MR的相互作用均显示出激活AP-1的潜力。消除GR和MR顺式激活激素反应元件驱动报告基因能力的DNA结合结构域突变,对这些反应的强度和极性产生了不同程度的影响。重要的是,MR对NFκB和AP-1信号传导的调节与反式机制一致,并且在原代人细胞中针对特定基因靶点证实了AP-1的作用。类固醇核受体对炎症信号传导的反式作用取决于具体情况,并受核受体构象、DNA序列和异源结合伴侣表达的影响。醛固酮对AP-1的激活可能有助于其在血管系统中的促炎作用。