title: Metabolic stability of superoxide adducts derived from newly developed cyclic nitrone spin traps.
abstract: Reactive oxygen species are by-products of aerobic metabolism involved in the onset and evolution of various pathological conditions. Among them, the superoxide radical is of special interest as the origin of several damaging species such as H2O2, hydroxyl radical, or peroxynitrite (ONOO(-)). Spin trapping coupled with ESR is a method of choice to characterize these species in chemical and biological systems and the metabolic stability of the spin adducts derived from reaction of superoxide and hydroxyl radicals with nitrones is the main limit to the in vivo application of the method. Recently, new cyclic nitrones bearing a triphenylphosphonium or permethylated β-cyclodextrin moiety have been synthesized and their spin adducts demonstrated increased stability in buffer. In this article, we studied the stability of the superoxide adducts of four new cyclic nitrones in the presence of liver subcellular fractions and biologically relevant reductants using an original setup combining a stopped-flow device and an ESR spectrometer. The kinetics of disappearance of the spin adducts were analyzed using an appropriate simulation program. Our results highlight the interest of the new spin trapping agents CD-DEPMPO and CD-DIPPMPO for specific detection of superoxide with high stability of the superoxide adducts in the presence of liver microsomes.
authors.affiliation: Laboratoire de Chimie et Biochimie Pharmacologiques et Toxicologiques (UMR CNRS 8601), Université Paris Descartes, 75006 Paris, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Bézière
authors.affiliation: Aix-Marseille Université, CNRS, ICR UMR 7273, 13397 Marseille Cedex 20, France.
authors.forename: Micael
authors.initials: M
authors.lastname: Hardy
authors.affiliation: Aix-Marseille Université, CNRS, ICR UMR 7273, 13397 Marseille Cedex 20, France.
authors.forename: Florent
authors.initials: F
authors.lastname: Poulhès
authors.affiliation: Aix-Marseille Université, CNRS, ICR UMR 7273, 13397 Marseille Cedex 20, France.
authors.forename: Hakim
authors.initials: H
authors.lastname: Karoui
authors.affiliation: Aix-Marseille Université, CNRS, ICR UMR 7273, 13397 Marseille Cedex 20, France.
authors.forename: Paul
authors.initials: P
authors.lastname: Tordo
authors.affiliation: Aix-Marseille Université, CNRS, ICR UMR 7273, 13397 Marseille Cedex 20, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Ouari
authors.affiliation: Laboratoire de Chimie et Biochimie Pharmacologiques et Toxicologiques (UMR CNRS 8601), Université Paris Descartes, 75006 Paris, France.
authors.forename: Yves-Michel
authors.initials: YM
authors.lastname: Frapart
authors.affiliation: Institute of Molecular Pharmacology, Research Centre for Natural Sciences, Budapest, Hungary.
authors.forename: Antal
authors.initials: A
authors.lastname: Rockenbauer
authors.affiliation: Laboratoire de Chimie et Biochimie Pharmacologiques et Toxicologiques (UMR CNRS 8601), Université Paris Descartes, 75006 Paris, France.
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Boucher
authors.affiliation: Laboratoire de Chimie et Biochimie Pharmacologiques et Toxicologiques (UMR CNRS 8601), Université Paris Descartes, 75006 Paris, France.
authors.forename: Daniel
authors.initials: D
authors.lastname: Mansuy
authors.affiliation: Laboratoire de Chimie et Biochimie Pharmacologiques et Toxicologiques (UMR CNRS 8601), Université Paris Descartes, 75006 Paris, France; IUFM de Paris, Université Paris Sorbonne, 75016 Paris, France. Electronic address: fabienne.peyrot@parisdescartes.fr.
authors.forename: Fabienne
authors.initials: F
authors.lastname: Peyrot
mesh_terms: D000818:Animals; D003600:Cytosol / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D022781:Hepatocytes / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000737:chemistry* / Q000378:metabolism; D017665:Hydroxyl Radical / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000737:chemistry; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism; D009589:Nitrogen Oxides / Q000737:chemistry*; D030421:Peroxynitrous Acid / Q000737:chemistry* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013113:Spin Labels; D019097:Spin Trapping; D013237:Stereoisomerism; D013481:Superoxides / Q000737:chemistry* / Q000378:metabolism
chemical_list: D009589:Nitrogen Oxides; D013113:Spin Labels; C015388:nitrones; D013481:Superoxides; D030421:Peroxynitrous Acid; D017665:Hydroxyl Radical; D006861:Hydrogen Peroxide
keywords: Cyclic nitrones; EPR spectroscopy; ESR spectroscopy; Free radicals; Hydroxyl radical; Spin adduct stability; Spin trapping; Superoxide