Medicines Evaluation Unit, National Institute for Health Research Translational Research Facility, University Hospital of South Manchester Foundation Trust, University of Manchester, Manchester, United Kingdom.
J Pharmacol Exp Ther. 2011 Sep;338(3):732-40. doi: 10.1124/jpet.111.180737. Epub 2011 May 24.
Corticosteroids partially suppress cytokine production by chronic obstructive pulmonary disease (COPD) alveolar macrophages. p38 mitogen-activated protein kinase (MAPK) inhibitors are a novel class of anti-inflammatory drug. We have studied the effects of combined treatment with a corticosteroid and a p38 MAPK inhibitor on cytokine production by COPD alveolar macrophages, with the aim of investigating dose-sparing and efficacy-enhancing effects. Alveolar macrophages from 10 patients with COPD, six smokers, and six nonsmokers were stimulated with lipopolysaccharide (LPS) after preincubation with five concentrations of dexamethasone alone, five concentrations of the p38 MAPK inhibitor 1-(5-tert-butyl-2-p-tolyl-2H-pyrazol-3-yl)-3(4-(2-morpholin-4-yl-ethoxy)naphthalen-1-yl)urea (BIRB-796) alone, and all combinations of these concentrations. After 24 h, the supernatants were analyzed for interleukin (IL)-8, IL-6, tumor necrosis factor α (TNFα), granulocyte macrophage-colony-stimulating factor (GM-CSF), IL-1α, IL-1β, IL-1ra, IL-10, monocyte chemoattractant protein 3, macrophage-derived chemokine (MDC), and regulated on activation normal T cell expressed and secreted (RANTES). The effect of dexamethasone on p38 MAPK activation was analyzed by Western blotting. Dexamethasone and BIRB-796 both reduced LPS-induced cytokine production in a dose-dependent manner in all subject groups, with no differences between groups. Increasing the concentration of BIRB-796 in combination with dexamethasone produced progressively greater inhibition of cytokine production than dexamethasone alone. There were significant efficacy-enhancing benefits and synergistic dose-sparing effects (p < 0.05) for the combination treatment for IL-8, IL-6, TNFα, GM-CSF, IL-1ra, IL-10, MDC, and RANTES in one or more subject groups. Dexamethasone had no effect on LPS-induced p38 MAPK activation. We conclude that p38 MAPK activation in alveolar macrophages is corticosteroid-insensitive. Combining a p38 MAPK inhibitor with a corticosteroid synergistically enhances the anti-inflammatory effects on LPS-mediated cytokine production by alveolar macrophages from patients with COPD and controls.
皮质类固醇可部分抑制慢性阻塞性肺疾病（COPD）肺泡巨噬细胞产生细胞因子。p38 丝裂原活化蛋白激酶（MAPK）抑制剂是一类新型抗炎药物。我们研究了皮质类固醇和 p38 MAPK 抑制剂联合治疗对 COPD 肺泡巨噬细胞产生细胞因子的影响，旨在研究剂量节省和疗效增强的效果。用脂多糖（LPS）刺激 10 名 COPD 患者、6 名吸烟者和 6 名不吸烟者的肺泡巨噬细胞，在单独预孵育五种浓度的地塞米松、五种浓度的 p38 MAPK 抑制剂 1-（5-叔丁基-2-对甲苯基-2H-吡唑-3-基）-3-（4-（2-吗啉-4-基-乙氧基）萘-1-基）脲（BIRB-796）和这些浓度的所有组合之后。24 小时后，分析上清液中白细胞介素（IL）-8、IL-6、肿瘤坏死因子-α（TNFα）、粒细胞-巨噬细胞集落刺激因子（GM-CSF）、IL-1α、IL-1β、IL-1ra、IL-10、单核细胞趋化蛋白 3、巨噬细胞衍生趋化因子（MDC）和调节激活正常 T 细胞表达和分泌（RANTES）。通过 Western 印迹分析地塞米松对 p38 MAPK 激活的影响。地塞米松和 BIRB-796 均以剂量依赖性方式降低所有受试人群中 LPS 诱导的细胞因子产生，且各组之间无差异。联合应用 BIRB-796 增加浓度可产生比单独使用地塞米松更大的细胞因子产生抑制作用。在一个或多个受试人群中，联合治疗对 IL-8、IL-6、TNFα、GM-CSF、IL-1ra、IL-10、MDC 和 RANTES 的疗效增强作用和协同剂量节省作用（p < 0.05）。地塞米松对 LPS 诱导的 p38 MAPK 激活没有影响。我们得出结论，肺泡巨噬细胞中的 p38 MAPK 激活对皮质类固醇不敏感。联合使用 p38 MAPK 抑制剂和皮质类固醇可协同增强对 COPD 患者和对照者肺泡巨噬细胞 LPS 介导的细胞因子产生的抗炎作用。
