Platelet adhesion to human vascular endothelium is modulated by constitutive and cytokine induced nitric oxide.
OBJECTIVE
The aim was to study whether basal or cytokine stimulated generation of nitric oxide (NO) modulates platelet adhesion to human umbilical vein endothelial cells (HUVEC).


METHODS
The adhesion of 111In labelled human platelets to transfected HUVEC (SGHEC-7) was measured either alone or after incubation of SGHEC-7 cells for 18 h with interleukin-1 beta (IL-1 beta) and/or tumour necrosis factor alpha (TNF alpha). The activity of NO synthase in these cells was measured by formation of citrulline. The effects of dexamethasone (0.3 microM) and NG-monomethyl-L-arginine (L-NMMA, 100 microM) on these two variables were determined.


RESULTS
Stimulation of SGHEC-7 cells with IL-1 beta or TNF alpha (each at 1-30 ng.ml-1) caused them to express the inducible NO synthase, an effect that was prevented by dexamethasone. Platelet adhesion to unstimulated SGHEC-7 cells was < 0.1% (n = 3) and was increased to 0.7 (SEM 0.2)% by L-NMMA but was not affected by dexamethasone. Stimulation of the cells with IL-1 beta and TNF alpha increased platelet adhesion to a maximum of 2.2(0.4)%. This increase was enhanced by both dexamethasone and L-NMMA. The effect of L-NMMA was prevented by L-arginine.


CONCLUSIONS
Inhibition of NO synthesis by L-NMMA potentiates platelet adhesion to unstimulated SGHEC-7 cells, showing that basally released NO regulates platelet adhesion. Stimulation of SGHEC-7 cells by cytokines increases their adhesive properties but at the same time causes them to express the inducible NO synthase. Nitric oxide generated by this enzyme contributes to the modulation of the adhesive properties of the endothelial cells. Thus both constitutive and inducible NO synthases modulate endothelial cell thrombogenicity.
Radomski
M W
MW
Wellcome Research Laboratories, Beckenham, Kent.
Vallance
P
P
Whitley
G
G
Foxwell
N
N
Moncada
S
S
D000594:Amino Acid Oxidoreductases / Q000096:biosynthesis; D001120:Arginine / Q000031:analogs & derivatives / Q000494:pharmacology; D002478:Cells, Cultured; D016207:Cytokines / Q000494:pharmacology*; D003907:Dexamethasone / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology*; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology; D009569:Nitric Oxide / Q000502:physiology*; D019001:Nitric Oxide Synthase; D010973:Platelet Adhesiveness / Q000187:drug effects*; D013268:Stimulation, Chemical; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D019323:omega-N-Methylarginine
D016207:Cytokines; D007375:Interleukin-1; D014409:Tumor Necrosis Factor-alpha; D019323:omega-N-Methylarginine; D009569:Nitric Oxide; D003907:Dexamethasone; D001120:Arginine; D019001:Nitric Oxide Synthase; D000594:Amino Acid Oxidoreductases