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Am J Physiol Cell Physiol 274: C1040-C1046, 1998;
0363-6143/98 $5.00
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Vol. 274, Issue 4, C1040-C1046, April 1998

Oxidant-induced arachidonic acid release and impairment of fatty acid acylation in vascular smooth muscle cells

Agnès Cane, Michelyne Breton, Kamen Koumanov, Gilbert Béréziat, and Odile Colard

Centre National de la Recherche Scientifique Unité de Recherche Associée 1283, Service de Biochimie, Centre Hospitalier Universitaire Saint-Antoine, 75012 Paris, France

Oxidative damage, which plays a major role in the early stages of atherosclerosis, is associated with arachidonic acid (AA) release in vascular smooth muscle cells (VSMC) as in other cell types. In this study, H2O2 was used to investigate mechanisms of AA release from VSMC on oxidative stress. Cell treatment with H2O2 inhibited AA incorporation in an inverse relationship to prolonged H2O2-induced AA release. Identical kinetics of inhibition of AA incorporation and AA release were observed after cell treatment with AlF-4, a process not involving phospholipase A2 (PLA2) activation as recently described (A. Cane, M. Breton, G. Béréziat, and O. Colard. Biochem. Pharmacol. 53: 327-337, 1997). AA release was not specific, since oleic acid also increased in the extracellular medium of cells treated with H2O2 or AlF-4 as measured by gas chromatography-mass spectrometry. In contrast, AA and oleic acid cell content decreased after cell treatment. Oleoyl and arachidonoyl acyl-CoA synthases and acyltransferases, assayed using a cell-free system, were not significantly modified. In contrast, a good correlation was observed between decreases in AA acylation and cell ATP content. The decrease in ATP content is only partially accounted for by mitochondrial damage as assayed by rhodamine 123 assay. We conclude that oxidant-induced arachidonate release results from impairment of fatty acid esterification and that ATP availability is probably responsible for free AA accumulation on oxidative stress by preventing its reesterification and/or transmembrane transport.

hydrogen peroxide; aluminum fluoride; cell adenosine triphosphate content; A7r5 cells


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