title: The cytotoxicity of nitroxyl: possible implications for the pathophysiological role of NO.
abstract: In addition to the broad repertoire of regulatory functions nitric oxide (NO) serves in mammalian physiology, the L-arginine:NO pathway is also involved in numerous pathophysiological mechanisms. While NO itself may actually protect cells from the toxicity of reactive oxygen radicals in some cases, it has been suggested that reactive nitrogen oxide species formed from nitric oxide synthase (NOS) can be cytotoxic. In addition to NO, the one electron reduction product NO- has been proposed to be formed from NOS. We investigated the potential cytotoxic role of nitroxyl (NO-), using the nitroxyl donor Angelis's salt, (AS; sodium trioxodinitrate, Na2N2O3) as the source of NO-. As was found to be cytotoxic to Chinese hamster V79 lung fibroblast cells over a concentration range of 2-4 mM. The presence of equimolar ferricyanide (Fe(III)-(CN6)3-), which converts NO- to NO, afforded dramatic protection against AS-mediated cytotoxicity. Treatment of V79 cells with L-buthionine sulfoximine to reduce intracellular glutathione markedly enhanced AS cytotoxicity, which suggests that GSH is critical for cellular protection against the toxicity of NO-. Further experiments showed that low molecular weight transition metal complexes associated with the formation of reactive oxygen species are not involved in AS-mediated cytotoxicity since metal chelators had no effect. However, under aerobic conditions, AS was more toxic than under hypoxic conditions, suggesting that oxygen dramatically enhanced AS-mediated cytotoxicity. At a molecular level, AS exposure resulted in DNA double strand breaks in whole cells, and this effect was completely prevented by coincubation of cells with ferricyanide or Tempol. The data in this study suggest that nitroxyl may contribute to the cytotoxicity associated with an enhanced expression of the L-arginine:NO pathway under different biological conditions.
authors.affiliation: Tumor Biology Section, National Cancer Institute, Bethesda, Maryland 20892, USA.
authors.forename: D A
authors.initials: DA
authors.lastname: Wink
authors.forename: M
authors.initials: M
authors.lastname: Feelisch
authors.forename: J
authors.initials: J
authors.lastname: Fukuto
authors.forename: D
authors.initials: D
authors.lastname: Chistodoulou
authors.forename: D
authors.initials: D
authors.lastname: Jourd'heuil
authors.forename: M B
authors.initials: MB
authors.lastname: Grisham
authors.forename: Y
authors.initials: Y
authors.lastname: Vodovotz
authors.forename: J A
authors.initials: JA
authors.lastname: Cook
authors.forename: M
authors.initials: M
authors.lastname: Krishna
authors.forename: W G
authors.initials: WG
authors.lastname: DeGraff
authors.forename: S
authors.initials: S
authors.lastname: Kim
authors.forename: J
authors.initials: J
authors.lastname: Gamson
authors.forename: J B
authors.initials: JB
authors.lastname: Mitchell
mesh_terms: D000818:Animals; D001120:Arginine / Q000378:metabolism; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D016923:Cell Death; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D003497:Cyclic N-Oxides / Q000494:pharmacology; D004249:DNA Damage; D005292:Ferricyanides / Q000494:pharmacology; D005347:Fibroblasts; D016166:Free Radical Scavengers; D005609:Free Radicals; D005978:Glutathione / Q000378:metabolism; D008168:Lung; D009569:Nitric Oxide / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000378:metabolism; D009573:Nitrites / Q000494:pharmacology; D009589:Nitrogen Oxides / Q000633:toxicity*; D013113:Spin Labels
chemical_list: D003497:Cyclic N-Oxides; D005292:Ferricyanides; D016166:Free Radical Scavengers; D005609:Free Radicals; D009573:Nitrites; D009589:Nitrogen Oxides; D013113:Spin Labels; C007931:hexacyanoferrate III; C021229:oxyhyponitrite; D009569:Nitric Oxide; D019328:Buthionine Sulfoximine; D001120:Arginine; D019001:Nitric Oxide Synthase; D005978:Glutathione; C039900:nitroxyl; C001803:tempol