title: Nitric oxide during ischemia attenuates oxidant stress and cell death during ischemia and reperfusion in cardiomyocytes.
abstract: Nitric oxide (NO) has been implicated as a cardioprotective agent during ischemia/reperfusion (I/R), but the mechanism of protection is unknown. Oxidant stress contributes to cell death in I/R, so we tested whether NO protects by attenuating oxidant stress. Cardiomyocytes and murine embryonic fibroblasts were administered NO (10-1200 nM) during simulated ischemia, and cell death was assessed during reperfusion without NO. In each case, NO abrogated cell death during reperfusion. Cells overexpressing endothelial NO synthase (NOS) exhibited a similar protection, which was abolished by the NOS inhibitor N(omega)-nitro-l-arginine methyl ester. Protection was not mediated by guanylate cyclase or the mitochondrial K(ATP) channel, as inhibitors of these systems failed to abolish protection. NO did not prevent decreases in mitochondrial potential, but cells protected with NO demonstrated recovery of potential at reperfusion. Measurements using C11-BODIPY reveal that NO attenuates lipid peroxidation during ischemia and reperfusion. Measurements of oxidant stress using the ratiometric redox sensor HSP-FRET demonstrate that NO attenuates protein oxidation during ischemia. These findings reveal that physiological levels of NO during ischemia can attenuate oxidant stress both during ischemia and during reperfusion. This response is associated with a remarkable attenuation of cell death, suggesting that ischemic cell death may be a regulated event.
authors.affiliation: Department of Medicine, University of Chicago, Chicago, IL 60637, USA.
authors.forename: Hirotaro
authors.initials: H
authors.lastname: Iwase
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Robin
authors.forename: Robert D
authors.initials: RD
authors.lastname: Guzy
authors.forename: Paul T
authors.initials: PT
authors.lastname: Mungai
authors.forename: Terry L
authors.initials: TL
authors.lastname: Vanden Hoek
authors.forename: Navdeep S
authors.initials: NS
authors.lastname: Chandel
authors.forename: Jacques
authors.initials: J
authors.lastname: Levraut
authors.forename: Paul T
authors.initials: PT
authors.lastname: Schumacker
mesh_terms: D000818:Animals; D016923:Cell Death / Q000502:physiology*; D002478:Cells, Cultured; D002642:Chick Embryo; D015227:Lipid Peroxidation / Q000502:physiology; D053078:Membrane Potential, Mitochondrial / Q000502:physiology; D008856:Microscopy, Fluorescence; D008929:Mitochondria, Heart / Q000378:metabolism / Q000473:pathology; D032383:Myocytes, Cardiac / Q000378:metabolism* / Q000473:pathology; D009569:Nitric Oxide / Q000378:metabolism*; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D018384:Oxidative Stress / Q000502:physiology*; D015427:Reperfusion Injury / Q000378:metabolism*
chemical_list: D009569:Nitric Oxide; D052250:Nitric Oxide Synthase Type III