Kent Lauren M, Smyth Lucy J C, Plumb Jonathan, Clayton Chris L, Fox Steve M, Ray David W, Farrow Stuart N, Singh Dave
Education and Research Centre (2nd Floor Office), South Manchester University Hospitals Trust, Wythenshawe Hospital, Southmoor Road, Manchester M23 9LT, UK.
J Pharmacol Exp Ther. 2009 Feb;328(2):458-68. doi: 10.1124/jpet.108.142950. Epub 2008 Nov 12.
p38 mitogen-activated protein kinase (MAPK) signaling is known to be increased in chronic obstructive pulmonary disease (COPD) macrophages. We have studied the effects of the p38 MAPK inhibitor N-cyano-N'-(2-{[8-(2,6-difluorophenyl)-4-(4-fluoro-2-methylphenyl)-7-oxo-7,8-dihydropyrido[2,3-d]-pyrimidin-2-yl]amino}ethyl)guanidine (SB706504) and dexamethasone on COPD macrophage inflammatory gene expression and protein secretion. We also studied the effects of combined SB706504 and dexamethasone treatment. Lipopolysaccharide (LPS)-stimulated monocyte derived macrophages (MDMs) and alveolar macrophages (AMs) were cultured with dexamethasone and/or SB706504. MDMs were used for gene array and protein studies, whereas tumor necrosis factor (TNF) alpha protein production was measured from AMs. SB706504 caused transcriptional inhibition of a range of cytokines and chemokines in COPD MDMs. The use of SB706504 combined with dexamethasone caused greater suppression of gene expression (-8.90) compared with SB706504 alone (-2.04) or dexamethasone (-3.39). Twenty-three genes were insensitive to the effects of both drugs, including interleukin (IL)-1beta, IL-18, and chemokine (CC motif) ligand (CCL) 5. In addition, the chromosome 4 chemokine cluster members, CXCL1, CXCL2, CXCL3, and CXCL8, were all glucocorticoid-resistant. SB706504 significantly inhibited LPS-stimulated TNFalpha production from COPD and smoker AMs, with near-maximal suppression caused by combination treatment with dexamethasone. We conclude that SB706504 targets a subset of inflammatory macrophage genes and when used with dexamethasone causes effective suppression of these genes. SB706504 and dexamethasone had no effect on the transcription of a subset of LPS-regulated genes, including IL-1beta, IL-18, and CCL5, which are all known to be involved in the pathogenesis of COPD.
已知p38丝裂原活化蛋白激酶（MAPK）信号通路在慢性阻塞性肺疾病（COPD）巨噬细胞中增强。我们研究了p38 MAPK抑制剂N-氰基-N'-（2-{[8-（2,6-二氟苯基）-4-（4-氟-2-甲基苯基）-7-氧代-7,8-二氢吡啶并[2,3-d]嘧啶-2-基]氨基}乙基）胍（SB706504）和地塞米松对COPD巨噬细胞炎症基因表达和蛋白分泌的影响。我们还研究了SB706504与地塞米松联合治疗的效果。用脂多糖（LPS）刺激单核细胞衍生巨噬细胞（MDM）和肺泡巨噬细胞（AM），并用地塞米松和/或SB706504进行培养。MDM用于基因芯片和蛋白研究，而从AM中检测肿瘤坏死因子（TNF）α蛋白的产生。SB706504导致COPD MDM中一系列细胞因子和趋化因子的转录抑制。与单独使用SB706504（-2.04）或地塞米松（-3.39）相比，SB706504与地塞米松联合使用对基因表达的抑制作用更强（-8.90）。23个基因对两种药物的作用均不敏感，包括白细胞介素（IL）-1β、IL-18和趋化因子（CC基序）配体（CCL）5。此外，4号染色体趋化因子簇成员CXCL1、CXCL2、CXCL3和CXCL8均对糖皮质激素耐药。SB706504显著抑制LPS刺激的COPD和吸烟者AM中TNFα的产生，与地塞米松联合治疗可产生近乎最大程度的抑制。我们得出结论，SB706504靶向炎症巨噬细胞基因的一个子集，与地塞米松联合使用时可有效抑制这些基因。SB706504和地塞米松对LPS调节基因的一个子集的转录没有影响，这些基因包括IL-1β、IL-18和CCL5，它们均参与COPD的发病机制。
