title: Inhibition of l-arginine transport in platelets by asymmetric dimethylarginine and N-monomethyl-l-arginine: effects of arterial hypertension.
abstract: Nitric oxide (NO) produced by human platelets plays an important role in all stages of platelet activation. l-Arginine, the precursor for NO synthesis, modulates NO production by platelets. The l-arginine analogues asymmetric dimethylarginine (ADMA) and N(G)-monomethyl-l-arginine (l-NMMA) are endogenous inhibitors of nitric oxide synthase (NOS), involved in the physiopathology of arterial hypertension. The aim of the present study was to investigate the inhibitory effects of endogenous and exogenous l-arginine analogues on l-arginine influx in platelets from healthy controls and hypertensive patients. Twelve patients with uncomplicated essential hypertension (stage I) and 15 age-matched normotensive controls participated in the present study. Platelets were isolated and incubated with l-[(3)H]-arginine and increasing concentrations of l-arginine analogues (5-2000 micromol/L). The influx of l-arginine was inhibited in a concentration-dependent manner by l-NMMA in platelets from controls (K(i) = 42 +/- 6 micromol/L) and this inhibitory effect was markedly higher in hypertensive platelets (K(i) = 23 +/- 4 micromol/L). Similarly, the K(i) for ADMA inhibition of l-arginine transport was significantly more pronounced in platelets from hypertensive patients (K(i) = 16 +/- 1 micromol/L) compared with controls (K(i) = 27 +/- 2 micromol/L). In contrast, N(G)-nitro-l-arginine methyl ester (l-NAME) was found to be a weak inhibitor of l-arginine influx in platelets from controls (K(i) = 1917 +/- 319 micromol/L) and hypertensive patients (K(i) = 2279 +/- 578 micromol/L). Aminoguanidine, a selective inhibitor of inducible NOS, failed to inhibit l-arginine transport. Our findings provide the first evidence that ADMA and l-NMMA markedly inhibit l-arginine transport in human platelets, an effect that is more pronounced in hypertensive patients. It is possible that endogenous l-arginine analogues, by inhibiting NO synthesis, are involved in the platelet activation present in hypertension.
authors.affiliation: Departamento de Farmacologia e Psicobiologia, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Tmc
authors.initials: T
authors.lastname: Brunini
authors.forename: Mb
authors.initials: M
authors.lastname: Moss
authors.forename: Mas
authors.initials: M
authors.lastname: Siqueira
authors.forename: Lr
authors.initials: L
authors.lastname: Meirelles
authors.forename: Al
authors.initials: A
authors.lastname: Rozentul
authors.forename: Ge
authors.initials: G
authors.lastname: Mann
authors.forename: Jc
authors.initials: J
authors.lastname: Ellory
authors.forename: R
authors.initials: R
authors.lastname: Soares de Moura
authors.forename: Ac
authors.initials: A
authors.lastname: Mendes-Ribeiro
mesh_terms: D000328:Adult; D001120:Arginine / Q000031:analogs & derivatives* / Q000097:blood* / Q000494:pharmacology*; D001158:Arteries / Q000503:physiopathology; D001693:Biological Transport, Active / Q000187:drug effects; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005260:Female; D006801:Humans; D006973:Hypertension / Q000188:drug therapy* / Q000503:physiopathology; D008297:Male; D008875:Middle Aged; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors*; D052250:Nitric Oxide Synthase Type III; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism; D019323:omega-N-Methylarginine / Q000494:pharmacology*; D020245:p-Chloromercuribenzoic Acid / Q000494:pharmacology
chemical_list: D004791:Enzyme Inhibitors; C487735:dimethylarginine; D019323:omega-N-Methylarginine; D020245:p-Chloromercuribenzoic Acid; D001120:Arginine; C496314:NOS3 protein, human; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III