title: Exogenous Nitric Oxide Protects Human Embryonic Stem Cell-Derived Cardiomyocytes against Ischemia/Reperfusion Injury.
abstract: Background and Aims. Human embryonic stem cell- (hESC-) derived cardiomyocytes are one of the useful screening platforms of potential cardiocytoprotective molecules. However, little is known about the behavior of these cardiomyocytes in simulated ischemia/reperfusion conditions. In this study, we have tested the cytoprotective effect of an NO donor and the brain type natriuretic peptide (BNP) in a screening platform based first on differentiated embryonic bodies (EBs, 6 + 4 days) and then on more differentiated cardiomyocytes (6 + 24 days), both derived from hESCs. Methods. Both types of hESC-derived cells were exposed to 150 min simulated ischemia, followed by 120 min reperfusion. Cell viability was assessed by propidium iodide staining. The following treatments were applied during simulated ischemia in differentiated EBs: the NO-donor S-nitroso-N-acetylpenicillamine (SNAP) (10(-7), 10(-6), and 10(-5) M), BNP (10(-9), 10(-8), and 10(-7) M), and the nonspecific NO synthase inhibitor Nω-nitro-L-arginine (L-NNA, 10(-5) M). Results. SNAP (10(-6), 10(-5) M) significantly attenuated cell death in differentiated EBs. However, simulated ischemia/reperfusion-induced cell death was not affected by BNP or by L-NNA. In separate experiments, SNAP (10(-6) M) also protected hESC-derived cardiomyocytes. Conclusions. We conclude that SNAP, but not BNP, protects differentiated EBs or cardiomyocytes derived from hESCs against simulated ischemia/reperfusion injury. The present screening platform is a useful tool for discovery of cardiocytoprotective molecules and their cellular mechanisms.
authors.affiliation: Cardiovascular Research Group, Department of Biochemistry, Faculty of Medicine, University of Szeged, Szeged 6720, Hungary.
authors.forename: János
authors.initials: J
authors.lastname: Pálóczi
authors.affiliation: Cardiovascular Research Group, Department of Biochemistry, Faculty of Medicine, University of Szeged, Szeged 6720, Hungary; Department of Pharmacology and Pharmacotherapy, Semmelweis University, Budapest 1085, Hungary.
authors.forename: Zoltán V
authors.initials: ZV
authors.lastname: Varga
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest 1519, Hungary.
authors.forename: Ágota
authors.identifier: 0000-0003-0380-8139
authors.initials: Á
authors.lastname: Apáti
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest 1519, Hungary.
authors.forename: Kornélia
authors.initials: K
authors.lastname: Szebényi
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest 1519, Hungary; MTA-SE Molecular Biophysics Research Group, Semmelweis University, Institute of Biophysics and Radiation Biology, Budapest 1085, Hungary.
authors.forename: Balázs
authors.initials: B
authors.lastname: Sarkadi
authors.affiliation: Department of Neuroscience and Imaging, Center of Excellence on Aging, "G. d'Annunzio" University, 66100 Chieti, Italy.
authors.forename: Rosalinda
authors.initials: R
authors.lastname: Madonna
authors.affiliation: Department of Neuroscience and Imaging, Center of Excellence on Aging, "G. d'Annunzio" University, 66100 Chieti, Italy.
authors.forename: Raffaele
authors.initials: R
authors.lastname: De Caterina
authors.affiliation: Cardiovascular Research Group, Department of Biochemistry, Faculty of Medicine, University of Szeged, Szeged 6720, Hungary.
authors.forename: Tamás
authors.initials: T
authors.lastname: Csont
authors.affiliation: Department of Experimental Pharmacology and Toxicology, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Eschenhagen
authors.affiliation: Cardiovascular Research Group, Department of Biochemistry, Faculty of Medicine, University of Szeged, Szeged 6720, Hungary; Department of Pharmacology and Pharmacotherapy, Semmelweis University, Budapest 1085, Hungary; Pharmahungary Group, Szeged 6722, Hungary.
authors.forename: Péter
authors.initials: P
authors.lastname: Ferdinandy
authors.affiliation: Cardiovascular Research Group, Department of Biochemistry, Faculty of Medicine, University of Szeged, Szeged 6720, Hungary; Department of Pharmacology and Pharmacotherapy, Semmelweis University, Budapest 1085, Hungary; Pharmahungary Group, Szeged 6722, Hungary.
authors.forename: Anikó
authors.initials: A
authors.lastname: Görbe
mesh_terms: D001120:Arginine / Q000494:pharmacology; D002316:Cardiotonic Agents / Q000494:pharmacology / Q000627:therapeutic use*; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D019610:Cytoprotection / Q000187:drug effects*; D000066449:Human Embryonic Stem Cells / Q000473:pathology*; D006801:Humans; D015428:Myocardial Reperfusion Injury / Q000188:drug therapy* / Q000473:pathology*; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000473:pathology*; D020097:Natriuretic Peptide, Brain / Q000494:pharmacology; D009569:Nitric Oxide / Q000494:pharmacology* / Q000627:therapeutic use*; D060888:Real-Time Polymerase Chain Reaction; D026423:S-Nitroso-N-Acetylpenicillamine / Q000494:pharmacology; D019210:Troponin I / Q000378:metabolism
chemical_list: D002316:Cardiotonic Agents; D019210:Troponin I; D020097:Natriuretic Peptide, Brain; D009569:Nitric Oxide; D026423:S-Nitroso-N-Acetylpenicillamine; D001120:Arginine