Cyclooxygenase inhibition decreases nitric oxide synthase activity in human platelets.
Activity of both nitric oxide (NO) synthase (NOS) and cyclooxygenase (COX) plays an important role in the regulation of platelet function. NO has been shown to directly activate COX. This study was designed to determine whether products of the COX pathway in turn regulate NOS activity. Human platelets were incubated with aspirin, indomethacin, the selective thromboxane A2 synthase inhibitor U-63557A, or the prostaglandin H2-thromboxane A2-receptor blocker SQ-29548 for 1 h at 37 degrees C. Multiple indexes of the activity of the L-arginine-NO pathway and changes in cytosolic Ca2+ concentration ([Ca2+]i) were measured in platelets. Both aspirin and indomethacin decreased NOS activity, measured as the conversion of L-arginine to L-citrulline and nitrite (+nitrate) formation, in platelets in a concentration-dependent fashion. Aspirin also decreased guanosine 3',5'-cyclic monophosphate accumulation in platelets. The NOS inhibitory effects of these aspirin and indomethacin effects were reversed by coincubation with the thromboxane A2 analog U-46619 or an excess of CaCl2. Incubation of COX inhibitors with platelets was associated with significant reductions in basal as well as thrombin-stimulated [Ca2+]i, and the reduction in [Ca2+]i was reversed by U-46619. Incubation of platelets with U-63557A and SQ-29548 resulted in inhibitory effects on NOS activity qualitatively similar to those of COX inhibitors. The effects of COX inhibitors or U-63557A were not associated with a change in NOS protein expression in platelets. These data suggest that NOS activity in human platelets is inhibited by COX inhibitors, mediated, at least in part, via suppression of thromboxane A2 and [Ca2+]i mobilization in platelets.
Chen
L
L
Department of Medicine, University of Florida and Veterans Affairs Medical Center, Gainesville 32610-0277, USA.
Salafranca
M N
MN
Mehta
J L
JL
D001241:Aspirin / Q000494:pharmacology; D001792:Blood Platelets / Q000187:drug effects* / Q000378:metabolism*; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D006152:Cyclic GMP / Q000097:blood; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology*; D003600:Cytosol / Q000378:metabolism; D006801:Humans; D007213:Indomethacin / Q000494:pharmacology; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors* / Q000097:blood; D009994:Osmolar Concentration; D013929:Thromboxane B2 / Q000097:blood
D016861:Cyclooxygenase Inhibitors; D013929:Thromboxane B2; D019001:Nitric Oxide Synthase; D006152:Cyclic GMP; D001241:Aspirin; D002118:Calcium; D007213:Indomethacin