Ethanol interferes with collagen-induced platelet activation by inhibition of arachidonic acid mobilization.
The effect of ethanol on signal generation in collagen-stimulated human platelets was evaluated. Incubation of washed human platelets with physiologically relevant concentrations of ethanol (25-150 mM) resulted in a dose-dependent inhibition of aggregation and secretion in response to collagen (0.5-10 micrograms/ml), but did not inhibit shape change. In platelets labeled with [3H]arachidonic acid, ethanol significantly inhibited the release of arachidonic acid from phospholipids, in both the presence and the absence of indomethacin. Thromboxane B2 formation was also inhibited in proportion to the reduction in free arachidonic acid. There was a close correlation between the extent of inhibition of arachidonic acid release and secretion. The inhibition of platelet aggregation and secretion by ethanol was partially overcome by the addition of exogenous arachidonic acid. In the presence of indomethacin, ethanol had no effect on the activation of phospholipase C by collagen as determined by the formation of inositol phosphates and phosphatidic acid. Moreover, ethanol had no effect on the mobilization of intracellular calcium by collagen and only minimally inhibited the early phases of the phosphorylation of myosin light chain (20 kDa) and a 47-kDa protein, a known substrate for protein kinase C. Arachidonic acid formation was also inhibited by ethanol in response to ionomycin under conditions where phospholipase C activation was prevented. The results suggest that the functional effects of ethanol on collagen-stimulated platelets are due, at least in part, to an inhibition of phospholipase A2.
Rubin
R
R
Department of Pathology and Cell Biology, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania 19107.
D000255:Adenosine Triphosphate / Q000378:metabolism; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000008:administration & dosage / Q000378:metabolism*; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism*; D003094:Collagen / Q000378:metabolism / Q000494:pharmacology* / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D000431:Ethanol / Q000494:pharmacology*; D006801:Humans; D007213:Indomethacin / Q000494:pharmacology; D007295:Inositol Phosphates / Q000096:biosynthesis; D010712:Phosphatidic Acids / Q000096:biosynthesis; D010741:Phospholipases A / Q000037:antagonists & inhibitors; D054467:Phospholipases A2; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology*; D010738:Type C Phospholipases / Q000378:metabolism
D001095:Arachidonic Acids; D007295:Inositol Phosphates; D010712:Phosphatidic Acids; D010975:Platelet Aggregation Inhibitors; D016718:Arachidonic Acid; D000431:Ethanol; D000255:Adenosine Triphosphate; D003094:Collagen; D010741:Phospholipases A; D054467:Phospholipases A2; D010738:Type C Phospholipases; D007213:Indomethacin