Aldosterone Abrogates Nuclear Factor κB-Mediated Tumor Necrosis Factor α Production in Human Neutrophils via the Mineralocorticoid Receptor
Aldosterone Abrogates Nuclear Factor κB-Mediated Tumor Necrosis Factor α Production in Human Neutrophils via the Mineralocorticoid Receptor
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DOI：
10.1161/hypertensionaha.109.141309
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发表时间：
2010-02-01
期刊：
影响因子：
8.3
通讯作者：
Kettritz, Ralph
中科院分区：
文献类型：
--
作者：
Bergmann, Astrid;Eulenberg, Claudia;Kettritz, Ralph
盐皮质激素受体（MR）通过醛固酮激活控制盐稳态和炎症在几种组织和细胞类型。多形核白细胞中是否存在功能性MR尚不清楚。我们研究了醛固酮通过MR调节炎性中性粒细胞反应的假设。通过流式细胞术、蛋白质印迹分析和显微镜检查，我们发现中性粒细胞具有MR。用醛固酮（10（-11）至10（-6）M）预孵育可剂量依赖性地抑制白细胞介素（IL）-8和粒细胞/巨噬细胞集落刺激因子（GM-CSF）中核因子κ B的活化。通过I κ B α蛋白质印迹法、电泳迁移率变动测定法和I κ B α mRNA的RT-PCR，在纤连蛋白上处理中性粒细胞。醛固酮对肿瘤坏死因子α和脂多糖介导的核因子κ B活化或IL-8和粒细胞/巨噬细胞集落刺激因子诱导的细胞外信号调节激酶、p38丝裂原活化蛋白激酶或磷脂酰肌醇3激酶/Akt活化无影响。螺内酯阻止核因子κ B抑制，表明MR特异性醛固酮效应。通过RT-PCR，我们发现中性粒细胞具有11 β-羟类固醇脱氢酶。在IL-8处理的细胞上清液中，由核因子κ B控制的肿瘤坏死因子α增加。醛固酮完全阻止了这种作用。RT-PCR显示，IL-8可使肿瘤坏死因子α mRNA显著增加，但可被醛固酮阻断，排除了肿瘤坏死因子α增加仅是分泌所致的可能性。最后，IL-8处理的中性粒细胞的条件培养基增加内皮细胞上的细胞间粘附分子-1的表达，随后IL-8处理的中性粒细胞粘附到内皮细胞。这些影响减少时，条件培养基从醛固酮预处理的中性粒细胞使用，和螺内酯阻断醛固酮的作用。我们的数据表明，一个功能性MR存在于中性粒细胞介导的炎症反应，是在工作时，中性粒细胞与内皮细胞相互作用。这些数据可能与MR阻断治疗方案相关。（高血压。2010;55：370-379）。
Mineralocorticoid receptor (MR) activation by aldosterone controls salt homeostasis and inflammation in several tissues and cell types. Whether or not a functional MR exists in polymorphonuclear neutrophils is unknown. We investigated the hypothesis that aldosterone modulates inflammatory neutrophil responses via the MR. By flow cytometry, Western blot analysis, and microscopy, we found that neutrophils possess MR. Preincubation with aldosterone (10(-11) to 10(-6) M) dose-dependently inhibited nuclear factor kappa B activation in interleukin (IL)-8- and granulocyte/macrophage colony-stimulating factor-treated neutrophils on fibronectin by I kappa B alpha Western blotting, electrophoretic mobility shift assay, and RT-PCR for I kappa B alpha mRNA. Aldosterone had no effect on tumor necrosis factor alpha- and lipopolysaccharide-mediated nuclear factor kappa B activation or on IL-8- and granulocyte/macrophage colony-stimulating factor-induced extracellular signal-regulated kinase, p38 mitogen-activated protein kinase, or phosphatidylinositol 3-kinase/Akt activation. Spironolactone prevented nuclear factor kappa B inhibition, indicating an MR-specific aldosterone effect. By RT-PCR, we found that neutrophils have 11 beta-hydroxysteroid dehydrogenase. Tumor necrosis factor alpha, which is controlled by nuclear factor kappa B, increased in the cell supernatant with IL-8 treatment. Aldosterone completely prevented this effect. RT-PCR showed a strong tumor necrosis factor alpha mRNA increase with IL-8 that was blocked by aldosterone, excluding the possibility that the tumor necrosis factor alpha increase was merely a consequence of secretion. Finally, conditioned medium from IL-8-treated neutrophils increased intercellular adhesion molecule-1 expression on endothelial cells and subsequently the adhesion of IL-8-treated neutrophils to endothelial cells. These effects were reduced when conditioned medium from aldosterone-pretreated neutrophils was used, and spironolactone blocked the aldosterone effect. Our data indicate that a functional MR exists in neutrophils mediating antiinflammatory effects that are at work when neutrophils interact with endothelial cells. These data could be relevant to MR-blockade treatment protocols. (Hypertension. 2010;55:370-379.)