Involvement of p38 and p42/44 MAP kinases and protein kinase C in the interferon-gamma and interleukin-1alpha-induced phosphorylation of 85-kDa cytosolic phospholipase A(2) in primary human bronchial epithelial cells.
Interferon-gamma (IFN-gamma) and interleukin-1 (IL-1) play an important role in the modulation of acute and chronic airway inflammation. Both IFN-gamma and IL-1 are known to increase the release of arachidonic acid (AA) from airway epithelial cells, suggesting that AA metabolites may mediate the cytokine-induced inflammation. This study was designed to examine the direct effect of IFN-gamma and IL-alpha on the phosphorylation of 85-kDa cytosolic phospholipase A(2) (cPLA(2)) and AA release in primary normal human bronchial epithelial (NHBE) cells. Treatment with IFN-gamma and IL-1alpha for 15 min induced a rapid increase of AA release from NHBE cells, which was blocked by the cPLA(2) inhibitor MAFP (p<0.05) but not by the sPLA(2) inhibitor LY311727 or iPLA(2) inhibitor HELSS. Immunoprecipitation and Western blot analysis showed that both IFN-gamma and IL-1alpha induced a rapid phosphorylation of cPLA(2). The IFN-gamma and IL-1alpha-induced cPLA(2) phosphorylation and AA release in the NHBE cells were inhibited by the p38 MAP kinase (MAPK) inhibitor SB203580, p42/44 MAPK inhibitor PD98059 and protein kinase C (PKC) inhibitor bisindolylmaleimide I. These results demonstrate the involvement of p38 and p42/44 MAPKs as well as PKC in the IFN-gamma and IL-1alpha-induced cPLA(2) phosphorylation and AA release in human airway epithelial cells.
Wu
Tong
T
Department of Pathology, University of Pittsburgh School of Medicine, Presbyterian University Hospital C902, 200 Lothrop Street, Pittsburgh, PA 15213, USA. wut@msx.upmc.edu
Han
Chang
C
Shelhamer
James H
JH
D016718:Arachidonic Acid / Q000378:metabolism; D001095:Arachidonic Acids / Q000494:pharmacology; D001980:Bronchi / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D005419:Flavonoids / Q000494:pharmacology; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D007371:Interferon-gamma / Q000494:pharmacology*; D007375:Interleukin-1 / Q000494:pharmacology*; D019950:Mitogen-Activated Protein Kinase 1 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009281:Naphthalenes / Q000494:pharmacology; D063065:Organophosphonates / Q000494:pharmacology; D010741:Phospholipases A / Q000037:antagonists & inhibitors / Q000187:drug effects / Q000378:metabolism*; D010766:Phosphorylation; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000187:drug effects / Q000378:metabolism*; D011725:Pyridines / Q000494:pharmacology; D011753:Pyrones / Q000494:pharmacology; D048051:p38 Mitogen-Activated Protein Kinases
D001095:Arachidonic Acids; D004791:Enzyme Inhibitors; D005419:Flavonoids; D007093:Imidazoles; D007375:Interleukin-1; D009281:Naphthalenes; D063065:Organophosphonates; D011725:Pyridines; D011753:Pyrones; C104810:methyl arachidonylfluorophosphonate; D016718:Arachidonic Acid; D007371:Interferon-gamma; C068314:6-(bromomethylene)tetrahydro-3-(1-naphthaleneyl)-2H-pyran-2-one; D011493:Protein Kinase C; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; D010741:Phospholipases A; C093642:SB 203580; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one