Arachidonic acid stimulates protein tyrosine phosphorylation in vascular cells. Buckley, Barbara J., and A. Richard Whorton. Departments of Medicine and Pharmacology, Duke University Medical Center, Durham, NC 27710
APStracts 2:0224C, 1995.
Arachidonic acid and its metabolites are important cellular mediators. In this study, we report a novel role for arachidonic acid in vascular cell signalling. We tested the effects of exogenous arachidonic acid on protein tyrosine phosphorylation in cultured vascular endothelial and smooth muscle cells. Arachidonic acid stimulated the phosphorylation of tyrosine-containing proteins of approximately 58, 93 and 120 kDa in the three cell types studied. This response was dose-dependent with a maximum effect observed with 40 NM arachidonic acid. Phosphorylation was rapid and transient, reaching a peak 0.5 min following the addition of arachidonic acid and returning to baseline by 8 min. A common set of protein substrates was phosphorylated in smooth muscle cells treated with the Ca2+-mobilizing agonist endothelin, concommitant with an increase in endogenous unesterified arachidonic acid. In order to determine whether the protein tyrosine phosphorylation was due to arachidonic acid or to a metabolite, we used inhibitors of cyclooxygenase, lipoxygenase and epoxygenase pathways. Ibuprofen, nordihydroguaiaretic acid, eicosatriynoic acid and eicosatetraynoic acid and 8-methoxypsoralen failed to inhibit the arachidonic acid -mediated response. We also found increased protein tyrosine phosphorylation following treatment with oleic, linolenic and [delta]-linoleic acid. These results suggest a mechanism of protein tyrosine phosphorylation that is directly stimulated by unmetabolized unsaturated fatty acids.

Received 6 March 1995; accepted in final form 26 May 1995.
APS Manuscript Number C122-5.
Article publication pending Am. J. Physiol. (Cell Physiology).
ISSN 1080-4757 Copyright 1995 The American Physiological Society.
Published in APStracts on  8 June 1995.