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Journal of Lipid Research, Vol. 42, 1467-1473, September 2001
Copyright © 2001 by Lipid Research, Inc.

Original Article

12(S)-Hydroperoxy-eicosatetraenoic acid increases arachidonic acid availability in collagen-primed platelets

Correspondence to: Catherine Calzada, To whom correspondence should be addressed., Catherine.Calzada{at}insa-lyon.fr (E-mail)

Lipid hydroperoxides have been reported to regulate cell function and eicosanoid formation. The aim of the present study was to determine the effect of 12(S)-hydroperoxy-eicosatetraenoic acid [12(S)-HPETE], the platelet 12-lipoxygenase-derived hydroperoxide of arachidonic acid (AA), on the availability of nonesterified AA, which represents a rate-limiting step in the biosynthesis of eicosanoids. The coincubation of human platelets with concentrations of 12(S)-HPETE below 50 nM and subthreshold concentrations (STC) of collagen (less than 0.24 µg/ml) significantly enhanced platelet aggregation and the formation of thromboxane B₂, the stable catabolite of the potent aggregating agent thromboxane A₂. In addition, the nonesterified endogenous AA concentration increased by 3-fold. Arachidonoyl-containing molecular species concentrations of 1,2-diacyl-glycero-3-phosphocholine, 1-alkyl-2-acyl-glycero-3-phosphocholine, and 1-alkenyl-2-acyl-glycero-3-phosphoethanolamine decreased specifically in response to 12(S)-HPETE, whereas no significant changes were observed within 1,2-diacyl-glycero-3-phosphoethanolamine and 1,2-diacyl-glycero-3-phosphoinositol molecular species. The 12(S)-HPETE-induced increase in nonesterified AA was fully prevented by arachidonoyl trifluoromethyl ketone, an inhibitor of cytosolic phospholipase A₂ (cPLA₂), and cPLA₂ was translocated to membranes and phosphorylated in platelets incubated with 12(S)-HPETE.

In conclusion, these results indicate that nanomolar concentrations of 12(S)-HPETE could play a significant role in controlling the level of endogenous AA and the formation of thromboxane, thereby potentiating platelet function. — Calzada, C., E. Véricel, B. Mitel, L. Coulon, and M. Lagarde. 12(S)-Hydroperoxy-eicosatetraenoic acid increases arachidonic acid availability in collagen-primed platelets. J. Lipid Res. 2001. 42: 1467;–1473.

Supplementary key words: cytosolic phospholipase A₂, hydroperoxides, phospholipid molecular species, platelet aggregation

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