title: Role of xanthine oxidoreductase in the anti-thrombotic effects of nitrite in rats in vivo.
abstract: The mechanisms underlying nitrite-induced effects on thrombosis and hemostasis in vivo are not clear. The goal of the work described here was to investigate the role of xanthine oxidoreductase (XOR) in the anti-platelet and anti-thrombotic activities of nitrite in rats in vivo. Arterial thrombosis was induced electrically in rats with renovascular hypertension by partial ligation of the left renal artery. Sodium nitrite (NaNO2, 0.17 mmol/kg twice daily for 3 days, p.o) was administered with or without one of the XOR-inhibitors: allopurinol (ALLO) and febuxostat (FEB) (100 and 5 mg/kg, p.o., for 3 days). Nitrite treatment (0.17 mmol/kg), which was associated with a significant increase in NOHb, nitrite/nitrate plasma concentration, resulted in a substantial decrease in thrombus weight (TW) (0.48 ± 0.03 mg vs. vehicle [VEH] 0.88 ± 0.08 mg, p < 0.001) without a significant hypotensive effect. The anti-thrombotic effect of nitrite was partially reversed by FEB (TW = 0.63 ± 0.06 mg, p < 0.05 vs. nitrites), but not by ALLO (TW = 0.43 ± 0.02 mg). In turn, profound anti-platelet effect of nitrite measured ex vivo using collagen-induced whole-blood platelet aggregation (70.5 ± 7.1% vs. VEH 100 ± 4.5%, p < 0.05) and dynamic thromboxaneB2 generation was fully reversed by both XOR-inhibitors. In addition, nitrite decreased plasminogen activator inhibitor-1 concentration (0.47 ± 0.13 ng/ml vs. VEH 0.62 ± 0.04 ng/ml, p < 0.05) and FEB/ALLO reversed this effect. In vitro the anti-platelet effect of nitrite (1 mM) was reversed by FEB (0.1 mM) under hypoxia (0.5%O2) and normoxia (20%O2). Nitrite treatment had no effect on coagulation parameters. In conclusion, the nitrite-induced anti-platelet effect in rats in vivo is mediated by XOR, but XOR does not fully account for the anti-thrombotic effects of nitrite.
authors.affiliation: a Department of Biopharmacy , Medical University of Bialystok , Bialystok , Poland .
authors.forename: K
authors.initials: K
authors.lastname: Kramkowski
authors.affiliation: a Department of Biopharmacy , Medical University of Bialystok , Bialystok , Poland .
authors.forename: A
authors.initials: A
authors.lastname: Leszczynska
authors.affiliation: c Jagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University , Krakow , Poland .
authors.forename: K
authors.initials: K
authors.lastname: Przyborowski
authors.affiliation: b Department of Pharmacodynamics , Medical University of Bialystok , Bialystok , Poland .
authors.forename: T
authors.initials: T
authors.lastname: Kaminski
authors.affiliation: a Department of Biopharmacy , Medical University of Bialystok , Bialystok , Poland .
authors.forename: U
authors.initials: U
authors.lastname: Rykaczewska
authors.affiliation: c Jagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University , Krakow , Poland .
authors.forename: B
authors.initials: B
authors.lastname: Sitek
authors.affiliation: c Jagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University , Krakow , Poland .
authors.forename: A
authors.initials: A
authors.lastname: Zakrzewska
authors.affiliation: c Jagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University , Krakow , Poland .
authors.forename: B
authors.initials: B
authors.lastname: Proniewski
authors.affiliation: d Department of Biochemistry , Medical University of Gdansk , Gdansk , Poland , and.
authors.forename: R T
authors.initials: RT
authors.lastname: Smolenski
authors.affiliation: a Department of Biopharmacy , Medical University of Bialystok , Bialystok , Poland .
authors.forename: E
authors.initials: E
authors.lastname: Chabielska
authors.affiliation: b Department of Pharmacodynamics , Medical University of Bialystok , Bialystok , Poland .
authors.forename: W
authors.initials: W
authors.lastname: Buczko
authors.affiliation: c Jagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University , Krakow , Poland .|e Department of Experimental Pharmacology , Chair of Pharmacology, Jagiellonian University Medical College , Krakow , Poland.
authors.forename: S
authors.initials: S
authors.lastname: Chlopicki
mesh_terms: D000818:Animals; D015415:Biomarkers; D001777:Blood Coagulation / Q000187:drug effects; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism; D001794:Blood Pressure / Q000187:drug effects; D004195:Disease Models, Animal; D005343:Fibrinolytic Agents / Q000008:administration & dosage / Q000493:pharmacokinetics / Q000494:pharmacology*; D006973:Hypertension / Q000097:blood / Q000188:drug therapy / Q000378:metabolism / Q000503:physiopathology; D008297:Male; D023421:Models, Animal; D009569:Nitric Oxide / Q000378:metabolism; D009573:Nitrites / Q000008:administration & dosage / Q000493:pharmacokinetics / Q000494:pharmacology*; D010974:Platelet Aggregation / Q000187:drug effects; D010979:Platelet Function Tests; D051381:Rats; D011957:Receptors, Opioid; D013927:Thrombosis / Q000097:blood / Q000188:drug therapy / Q000378:metabolism / Q000503:physiopathology; D014968:Xanthine Dehydrogenase / Q000378:metabolism*
chemical_list: D015415:Biomarkers; D005343:Fibrinolytic Agents; D009573:Nitrites; D011957:Receptors, Opioid; D009569:Nitric Oxide; D014968:Xanthine Dehydrogenase
keywords: Nitric oxide; nitrite; platelets; thrombosis; xanthine oxidoreductase