Effects of homocysteine on l-arginine transport and nitric oxide formation in human platelets.
BACKGROUND
Recent evidence indicates that hyperhomocysteinaemia is an independent risk factor for atherosclerosis, thrombosis and other cardiovascular diseases. This may be secondary to impaired fibrinolysis or increased platelet reactivity. Nitric oxide (NO), a product from l-arginine by NOS and potent antiaggregating agent, plays an important role in the regulation of platelet function.


METHODS
The present study aimed to define the effect of homocysteine on the l-arginine/NO pathway in human platelets. l-Arginine uptake, NO formation and Ca2+ levels were measured. Moreover the homocysteine effect on platelet activation induced by thrombin was tested.


RESULTS
Homocysteine causes a concentration-dependent inhibition of l-arginine transport. Results show that homocysteine does not modify the Km parameter, but it significantly decreases the Vmax value. The nitrite and nitrate formation, strictly correlated with the l-arginine transport, also significantly decreased. In contrast, cNOS activity remained unchanged upon homocysteine treatment. In addition homocysteine in a dose dependent manner increased the intracellular Ca2+ concentration and platelet response to thrombin.


CONCLUSIONS
Results indicate that the l-arginine/NO pathway is one of the various targets of homocysteine in human platelets. The increased Ca2+ levels associated with reduced NO formation may generate hyperactivation and may contribute to the thrombogenic processes.
Leoncini
G
G
Department of Experimental Medicine, University of Genova, Genova, Italy. leoncini@unige.it
Pascale
R
R
Signorello
M G
MG
D001120:Arginine / Q000378:metabolism*; D001692:Biological Transport / Q000187:drug effects; D001792:Blood Platelets / Q000187:drug effects* / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002474:Cell-Free System / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005033:Ethylmaleimide / Q000494:pharmacology; D006710:Homocysteine / Q000494:pharmacology*; D006801:Humans; D007930:Leucine / Q000494:pharmacology; D009569:Nitric Oxide / Q000096:biosynthesis*; D019001:Nitric Oxide Synthase / Q000378:metabolism; D010974:Platelet Aggregation / Q000502:physiology
D006710:Homocysteine; D009569:Nitric Oxide; D001120:Arginine; D019001:Nitric Oxide Synthase; D007930:Leucine; D005033:Ethylmaleimide; D002118:Calcium