Abstract
Corticosteroid insensitivity (CI) is a major barrier to treating severe asthma. Despite intensive research, the molecular mechanism of CI remains uncertain. The aim of this study was to determine abnormality in corticosteroid action in severe asthma and to identify the molecular mechanism of the long-acting β2-adrenergic agonists (LABAs) formoterol and salmeterol on restoration of corticosteroid sensitivity in severe asthma in vitro. Peripheral blood mononuclear cells (PBMCs) were obtained from 16 subjects with severe corticosteroid-insensitive asthma, 6 subjects with mild corticosteroid-sensitive asthma, and 11 healthy volunteers. Corticosteroid (dexamethasone) sensitivity was determined on tumor necrosis factor-α (TNF-α)-induced interleukin (IL)-8 production. Glucocorticoid receptor (GR) phosphorylation and kinase phosphorylation were evaluated by immunoprecipitation-Western blotting analysis and kinase phosphorylation array in IL-2/IL-4-treated corticosteroid insensitive model in PBMCs. In vitro corticosteroid sensitivity on TNF-α-induced IL-8 production was significantly lower in patients with severe asthma than in healthy volunteers and patients with mild asthma. This CI seen in severe asthma was associated with reduced GR nuclear translocation and with hyperphosphorylation of GR, which were reversed by LABAs. In IL-2/IL-4-treated PBMCs, LABAs inhibited phosphorylation of Jun-NH2-terminal kinase and p38 mitogen-activated protein kinase-γ (p38MAPK-γ) as well as GR. In addition, cells with p38MAPK-γ knockdown by RNA interference did not develop CI in the presence of IL-2/IL-4. Furthermore, p38MAPK-γ protein expression was up-regulated in PBMCs from some patients with severe asthma. In conclusion, p38 MAPK-γ activation impairs corticosteroid action and p38 MAPK-γ inhibition by LABAs has potential for the treatment of severe asthma.
Footnotes
↵ The online version of this article (available at http://molpharm.aspetjournals.org) contains supplemental material.
This study was supported by Asthma UK [Grant 04-56]; the UK Medical Research Council [Grant G0401662]; GlaxoSmithKline; and AstraZeneca (Lund).
Article, publication date, and citation information can be found at http://molpharm.aspetjournals.org.
doi:10.1124/mol.111.071993.
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ABBREVIATIONS:
- CI
- corticosteroid insensitivity
- GR
- glucocorticoid receptor
- NF-κB
- nuclear factor-κB
- JNK
- c-Jun-NH2-terminal kinase
- MAPK
- mitogen-activate protein kinase
- IL
- interleukin
- LABA
- long-acting β2-agonist
- SB203580
- 4-[4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-1H-imidazol-5-yl]pyridine
- FEV1
- forced expiratory volume in 1 s
- PBMC
- peripheral blood mononuclear cell
- FITC
- fluorescein isothiocyanate
- Dex
- dexamethasone
- PCR
- polymerase chain reaction
- ELISA
- enzyme-linked immunosorbent assay
- HSP
- heat shock protein
- KD
- knockdown
- FOR
- formoterol
- SAL
- salmeterol
- HV
- healthy volunteer(s)
- SA
- severe asthma
- MA
- mild glucocorticoid-sensitive asthma
- PP
- protein phosphatase.
- Received February 26, 2011.
- Accepted September 6, 2011.
- Copyright © 2011 The American Society for Pharmacology and Experimental Therapeutics
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