MEGAKARYOCYTES FROM PATIENTS WITH CORONARY ATHEROSCLEROSIS EXPRESS THE INDUCIBLE NITRIC-OXIDE SYNTHASE

Endothelial and platelet generation of nitric oxide (NO) plays an important role in the regulation of hemostasis. Alterations in NO biosynthesis are described in atherosclerosis. We have investigated the NO pathway in megakaryocytes and platelets from patients with atherosclerosis and age-matched control subjects. Megakaryocytes and platelets were isolated from patients with severe coronary atherosclerosis (n=19) and normal coronary arteries (n=9) as demonstrated by selective angiography. Constitutive (Ca2+-dependent) and inducible (Ca2+-independent) NO synthase (cNOS and iNOS, respectively) activities were measured by using the citrulline assay and by immunostaining techniques using an anti-peptide antibody to iNOS. Megakaryocytes from patients with atherosclerosis expressed significantly greater amounts of iNOS (1.28 +/- 0.46 pmol citrulline . mg(-1). min(-1)) than cNOS (0.29 +/- 0.40 pmol . mg(-1). min(-1)). In contrast, megakaryocytes from patients with normal coronary arteries expressed significantly more cNOS (1.48 +/- 0.23 pmol . mg(-1). min(-1)) than iNOS (0.49 +/- 0.40 pmol . mg(-1). min(-1)). Platelets isolated from both groups showed no significant difference in cNOS expression, and no iNOS was seen in either group. Immunostaining confirmed the presence of the iNOS in megakaryocytes. These results suggest there is a link between the expression of iNOS in the megakaryocyte and atherosclerosis.