title: Diminished nitric oxide generation from neutrophils suppresses platelet activation in chronic renal failure.
abstract: Chronic renal failure (CRF) is a complex clinical condition associated with accelerated atherosclerosis and thrombosis leading to cardiovascular events. The aim of this study was to investigate in detail the NO pathway in neutrophils obtained from hemodialysis patients and its association with platelet function and oxidative status. Fifteen CRF patients on hemodialysis and fifteen controls were included in this study. Laboratory and experimental evaluations were performed after hemodialysis in CRF patients. We evaluated L-[³H] arginine transport, NO synthase (NOS) activity, amino acid concentration in neutrophils, and expressions of NOS isoforms and p47(phox) by western blotting. Platelet aggregation was analyzed in the presence or absence of neutrophils. Oxidative status was measured through glutathione peroxidase, catalase activities, protein oxidation, lipid peroxidation, and DNA/RNA oxidation in serum. Basal NOS activity (pmol/10⁶ cells/min) was impaired in CRF patients on hemodialysis (0.33 ± 0.17) compared to controls (0.65 ± 0.12), whereas the expression of NOS isoforms remained unaltered. L-Arginine transport into neutrophils was similar in CRF patients on hemodialysis and controls. In addition, intracellular concentration of L-arginine was increased fourfold in the patient group. Systemic oxidative stress markers were not affected by CRF. On the other hand, NADPH oxidase subunit p47(phox) in neutrophils was overexpressed in CRF. In the presence of neutrophils, there was a reduction time-dependent in platelet aggregation in both groups with no difference between them. This data suggest that reduced basal generation of NO by neutrophils in CRF patients on hemodialysis occurs independently of L-arginine bioavailability and is able to suppress platelet activation.
authors.affiliation: Department of Pharmacology and Psychobiology, University of the State of Rio de Janeiro, Rio de Janeiro, 20551-030, Brazil.
authors.forename: Daniele C
authors.initials: DC
authors.lastname: Abrantes
authors.forename: Tatiana M C
authors.initials: TM
authors.lastname: Brunini
authors.forename: Cristiane
authors.initials: C
authors.lastname: Matsuura
authors.forename: Wanda Vianna
authors.initials: WV
authors.lastname: Mury
authors.forename: Carolina R
authors.initials: CR
authors.lastname: Corrêa
authors.forename: Sérgio F
authors.initials: SF
authors.lastname: Santos
authors.forename: Monique B O
authors.initials: MB
authors.lastname: Ormonde do Carmo
authors.forename: Antônio Cláudio
authors.initials: AC
authors.lastname: Mendes-Ribeiro
mesh_terms: D000328:Adult; D001120:Arginine / Q000097:blood; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D007676:Kidney Failure, Chronic / Q000097:blood* / Q000628:therapy; D008297:Male; D008875:Middle Aged; D009504:Neutrophils / Q000378:metabolism*; D009569:Nitric Oxide / Q000097:blood*; D019001:Nitric Oxide Synthase / Q000378:metabolism; D015539:Platelet Activation; D010974:Platelet Aggregation; D006435:Renal Dialysis
chemical_list: D009569:Nitric Oxide; D001120:Arginine; D019001:Nitric Oxide Synthase