title: Inhibitory effect of p38 mitogen-activated protein kinase inhibitors on cytokine release from human macrophages.
abstract: OBJECTIVE
  Macrophages release cytokines that may contribute to pulmonary inflammation in conditions such as chronic obstructive pulmonary disease. Thus, inhibition of macrophage cytokine production may have therapeutic benefit. p38 MAPK may regulate cytokine production, therefore, the effect of two p38 MAPK inhibitors, SB239063 and SD-282, on the release of TNF-alpha, GM-CSF and IL-8 from human macrophages was investigated.
  
  
  METHODS
  Cytokine release was measured by ELISA. Immunoblots and mRNA expression studies were performed to confirm p38 MAPK isoform expression and activity. Macrophages were isolated from lung tissue of current smokers, ex-smokers and emphysema patients and exposed to lipopolysaccharide. These cells then released cytokines in a concentration-dependent manner.
  
  
  RESULTS
  SB239063 only inhibited TNF-alpha release (EC50 0.3 +/- 0.1 microM). Disease status had no effect on the efficacy of SB239063. SD-282 inhibited both TNF-alpha and GM-CSF release from macrophages (EC50 6.1 +/- 1.4 nM and 1.8 +/- 0.6 microM respectively) but had no effect on IL-8 release. In contrast, both inhibitors suppressed cytokine production in monocytes.
  
  
  CONCLUSIONS
  The differential effects of p38 MAPK inhibitors between macrophages and monocytes could not be explained by differences in p38 MAPK isoform expression or activity. However, the stability of TNF-alpha mRNA was significantly increased in macrophages compared to monocytes. These data suggest a differential involvement for p38 MAPK in macrophage cytokine production compared with monocytes. These effects are not due to lack of p38 activation or p38alpha expression in macrophages but may reflect differential effects on the stability of cytokine mRNA.
authors.affiliation: Airway Disease Section, National Heart and Lung Institute, Imperial College London, London, UK.
authors.forename: S J
authors.initials: SJ
authors.lastname: Smith
authors.forename: P S
authors.initials: PS
authors.lastname: Fenwick
authors.forename: A G
authors.initials: AG
authors.lastname: Nicholson
authors.forename: F
authors.initials: F
authors.lastname: Kirschenbaum
authors.forename: T K
authors.initials: TK
authors.lastname: Finney-Hayward
authors.forename: L S
authors.initials: LS
authors.lastname: Higgins
authors.forename: M A
authors.initials: MA
authors.lastname: Giembycz
authors.forename: P J
authors.initials: PJ
authors.lastname: Barnes
authors.forename: L E
authors.initials: LE
authors.lastname: Donnelly
mesh_terms: D015153:Blotting, Western; D002478:Cells, Cultured; D016207:Cytokines / Q000187:drug effects / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005260:Female; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology* / Q000627:therapeutic use; D007211:Indoles / Q000494:pharmacology* / Q000627:therapeutic use; D016209:Interleukin-8 / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D016676:Macrophages, Alveolar / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D048291:Mitogen-Activated Protein Kinase 12 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D048308:Mitogen-Activated Protein Kinase 14 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D009000:Monocytes / Q000187:drug effects / Q000378:metabolism; D011014:Pneumonia / Q000188:drug therapy / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology* / Q000627:therapeutic use; D011743:Pyrimidines / Q000494:pharmacology* / Q000627:therapeutic use; D020871:RNA Stability / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000187:drug effects / Q000378:metabolism
chemical_list: C508606:CXCL8 protein, human; D016207:Cytokines; D007093:Imidazoles; D007211:Indoles; D016209:Interleukin-8; D008070:Lipopolysaccharides; D047428:Protein Kinase Inhibitors; D011743:Pyrimidines; D014409:Tumor Necrosis Factor-alpha; C514526:indole-5-carboxamide; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D048291:Mitogen-Activated Protein Kinase 12; D048308:Mitogen-Activated Protein Kinase 14; C406525:SB 239063