Abstract
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) α 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)-1β, 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 TNFα 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-1β, IL-18, and CCL5, which are all known to be involved in the pathogenesis of COPD.
Footnotes
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This work was supported by the Biotechnology and Biological Sciences Research Council and GlaxoSmithKline [Grant BBS/S/N/2004/11516].
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Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
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doi:10.1124/jpet.108.142950.
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ABBREVIATIONS: COPD, chronic obstructive pulmonary disease; AM, alveolar macrophage; GC, glucocorticoid; GR, glucocorticoid receptor; NF, nuclear factor; TLR, Toll-like receptor; LPS, lipopolysaccharide; MAPK, mitogen-activated protein kinase; TNF, tumor necrosis factor; IL, interleukin; GM-CSF, granulocyte macrophage colony-stimulating factor; MDM, monocyte-derived macrophage; PCR, polymerase chain reaction; UP, ultrapure; DMSO, dimethyl sulfoxide; qPCR, quantitative PCR; ELISA, enzyme-linked immunosorbent assay; IPA, ingenuity pathways analysis; FEV, forced expiratory volume; FVC, forced vital capacity; CCL, chemokine (CC motif) ligand; SB706504, 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; SB239063, trans-1-(4-hydroxycyclohexyl)-4-(4-fluorophenyl)-5-(2-methoxypyridimidin-4-yl)imidazole; SD-282, indole-5-carboxamide (ATP-competitive inhibitor of p38 kinase).
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↵ The online version of this article (available at http://jpet.aspetjournals.org) contains supplemental material.
- Received July 9, 2008.
- Accepted October 21, 2008.
- The American Society for Pharmacology and Experimental Therapeutics
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