title: Erythrocytes are the major intravascular storage sites of nitrite in human blood.
abstract: Plasma levels of nitrite ions have been used as an index of nitric oxide synthase (NOS) activity in vivo. Recent data suggest that nitrite is a potential intravascular repository for nitric oxide (NO), bioactivated by a nitrite reductase activity of deoxyhemoglobin. The precise levels and compartmentalization of nitrite within blood and erythrocytes have not been determined. Nitrite levels in whole blood and erythrocytes were determined using reductive chemiluminescence in conjunction with a ferricyanide-based hemoglobin oxidation assay to prevent nitrite destruction. This method yields sensitive and linear measurements of whole blood nitrite over 24 hours at room temperature. Nitrite levels measured in plasma, erythrocytes, and whole blood from 15 healthy volunteers were 121 plus or minus 9, 288 plus or minus 47, and 176 plus or minus 17 nM, indicating a surprisingly high concentration of nitrite within erythrocytes. The majority of nitrite in erythrocytes is located in the cytosol unbound to proteins. In humans, we found a significant artery-to-vein gradient of nitrite in whole blood and erythrocytes. Shear stress and acetylcholine-mediated stimulation of endothelial NOS significantly increased venous nitrite levels. These studies suggest a dynamic intravascular NO metabolism in which endothelial NOS-derived NO is stabilized as nitrite, transported by erythrocytes, and consumed during arterial-to-venous transit.
authors.affiliation: Molecular Medicine Branch, National Institute of Diabetes and Digestive and Kidney Disease, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: André
authors.initials: A
authors.lastname: Dejam
authors.forename: Christian J
authors.initials: CJ
authors.lastname: Hunter
authors.forename: Mildred M
authors.initials: MM
authors.lastname: Pelletier
authors.forename: Lewis L
authors.initials: LL
authors.lastname: Hsu
authors.forename: Roberto F
authors.initials: RF
authors.lastname: Machado
authors.forename: Sruti
authors.initials: S
authors.lastname: Shiva
authors.forename: Gordon G
authors.initials: GG
authors.lastname: Power
authors.forename: Malte
authors.initials: M
authors.lastname: Kelm
authors.forename: Mark T
authors.initials: MT
authors.lastname: Gladwin
authors.forename: Alan N
authors.initials: AN
authors.lastname: Schechter
mesh_terms: D000328:Adult; D004730:Endothelium, Vascular / Q000378:metabolism; D004912:Erythrocytes / Q000378:metabolism*; D005260:Female; D005292:Ferricyanides; D006454:Hemoglobins / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D008954:Models, Biological; D009569:Nitric Oxide / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III; D009573:Nitrites / Q000097:blood*; D010084:Oxidation-Reduction
chemical_list: D005292:Ferricyanides; D006454:Hemoglobins; D009573:Nitrites; C007931:hexacyanoferrate III; D009569:Nitric Oxide; C496314:NOS3 protein, human; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III