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J. Biol. Chem., Vol. 279, Issue 47, 48550-48561, November 19, 2004

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Cytosolic Phospholipase A₂ Group IV but Not Secreted Phospholipase A₂ Group IIA, V, or X Induces Interleukin-8 and Cyclooxygenase-2 Gene and Protein Expression through Peroxisome Proliferator-activated Receptors 1 and 2 in Human Lung Cells^*

Rafal Pawliczak, Carolea Logun, Patricia Madara, Marion Lawrence, Grzegorz Woszczek, Anetta Ptasinska, Marek L. Kowalski, Tong Wu¶, and James H. Shelhamer||

From the Critical Care Medicine Department, National Institutes of Health, Bethesda, Maryland 20892, the Department of Allergy and Clinical Immunology, Medical University of Lodz, Lodz 92213, Poland, and the ^¶Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213

It has been reported that interleukin-8 (IL-8) and cyclooxygenase-2 (COX-2) expression is regulated by peroxisome proliferator-activated receptor (PPAR)- synthetic ligands. We have shown previously that cytosolic phospholipase A₂ (cPLA₂) is able to activate gene expression through PPAR- response elements (Pawliczak, R., Han, C., Huang, X. L., Demetris, A. J., Shelhamer, J. H., and Wu, T. (2002) J. Biol. Chem. 277, 33153–33163). In this study we investigated the influence of cPLA₂ and secreted phospholipase A₂ (sPLA₂) Group IIA, Group V, and Group X on IL-8 and COX-2 expression in human lung epithelial cells (A549 cells). We also studied the results of cPLA₂ activation by epidermal growth factor (EGF) and calcium ionophore (A23187) on IL-8 and COX-2 reporter gene activity, mRNA level, and protein synthesis. cPLA₂ overexpression and activation increased both IL-8 and COX-2 reporter gene activity. Overexpression and activation of Group IIA, Group V, or Group X sPLA₂s did not increase IL-8 and COX-2 reporter gene activity. Methyl arachidonyl fluorophosphate, a cPLA₂ inhibitor, inhibited the effect of A23187 and of EGF on both IL-8 and COX-2 reporter gene activity, steady state levels of IL-8 and COX-2 mRNA, and IL-8 and COX-2 protein expression. Small inhibitory RNAs directed against PPAR-1 and -2 blunted the effect of A23187 and of EGF on IL-8 and COX-2 protein expression. Moreover small inhibitory RNAs directed against cPLA₂ decreased the effect of A23187 and EGF on IL-8 and COX-2 protein expression. These results demonstrate that cPLA₂ has an influence on IL-8 and COX 2 gene and protein expression at least in part through PPAR-.

Received for publication, August 4, 2004

^* This work was supported in part by Grant 3P04A02124 from the State Committee for Scientific Research, Poland (to R. P.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^|| To whom correspondence should be addressed: Critical Care Medicine Dept., National Institutes of Health, Warren Grant Magnuson Clinical Center, Bldg. 10, Rm. 7D43, 9000 Rockville Pike, Bethesda, MD 20892. E-mail: jshelhamer{at}nih.gov.

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