title: Early oxidative damage induced by doxorubicin: Source of production, protection by GKT137831 and effect on Ca(2+) transporters in HL-1 cardiomyocytes.
abstract: In atrial-derived HL-1 cells, ryanodine receptor and Na(+)/Ca(2+)-exchanger were altered early by 5 μM doxorubicin. The observed effects were an increase of cytosolic Ca(2+) at rest, ensuing ryanodine receptor phosphorylation, and the slowing of Ca(2+) transient decay after caffeine addition. Doxorubicin triggered a linear rise of reactive oxygen species (ROS) with no early effect on mitochondrial inner membrane potential. Doxorubicin and ROS were both detected in mitochondria by colocalization with fluorescence probes and doxorubicin-induced ROS was totally blocked by mitoTEMPO. The NADPH oxidase activity in the mitochondrial fraction was sensitive to inhibition by GKT137831, and doxorubicin-induced ROS decreased gradually as the GKT137831 concentration added in preincubation was increased. When doxorubicin-induced ROS was prevented by GKT137831, the kinetic response revealed a permanent degree of protection that was consistent with mitochondrial NADPH oxidase inhibition. In contrast, the ROS induction by doxorubicin after melatonin preincubation was totally eliminated at first but the effect was completely reversed with time. Limiting the source of ROS production is a better alternative for dealing with oxidative damage than using ROS scavengers. The short-term effect of doxorubicin on Ca(2+) transporters involved in myocardiac contractility was dependent on oxidative damage, and so the impairment was subsequent to ROS production.
authors.affiliation: Cardiología Clínica y Experimental, Departamento de Medicina Interna, Facultad de Medicina, Universidad de Murcia, Campus de El Palmar, 30120, Murcia, Spain.
authors.forename: Mari C
authors.initials: MC
authors.lastname: Asensio-López
authors.affiliation: Departamento de Bioquímica y Biología Molecular A, Universidad de Murcia, Campus de Espinardo, 30071, Murcia, Spain.
authors.forename: Fernando
authors.initials: F
authors.lastname: Soler
authors.affiliation: Cardiología Clínica y Experimental, Departamento de Medicina Interna, Facultad de Medicina, Universidad de Murcia, Campus de El Palmar, 30120, Murcia, Spain.
authors.forename: Jesús
authors.initials: J
authors.lastname: Sánchez-Más
authors.affiliation: Cardiología Clínica y Experimental, Departamento de Medicina Interna, Facultad de Medicina, Universidad de Murcia, Campus de El Palmar, 30120, Murcia, Spain; Servicio de Cardiología, Hospital Clínico Universitario Virgen de la Arrixaca, 30120, El Palmar, Murcia, Spain.
authors.forename: Domingo
authors.initials: D
authors.lastname: Pascual-Figal
authors.affiliation: Departamento de Bioquímica y Biología Molecular A, Universidad de Murcia, Campus de Espinardo, 30071, Murcia, Spain. Electronic address: fbelda@um.es.
authors.forename: Francisco
authors.initials: F
authors.lastname: Fernández-Belda
authors.affiliation: Cardiología Clínica y Experimental, Departamento de Medicina Interna, Facultad de Medicina, Universidad de Murcia, Campus de El Palmar, 30120, Murcia, Spain.
authors.forename: Antonio
authors.initials: A
authors.lastname: Lax
mesh_terms: D002118:Calcium / Q000378:metabolism*; D027682:Cation Transport Proteins / Q000378:metabolism*; D002460:Cell Line; D019610:Cytoprotection / Q000187:drug effects*; D004317:Doxorubicin / Q000494:pharmacology*; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D009200:Myocardial Contraction / Q000187:drug effects; D032383:Myocytes, Cardiac / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects*; D011720:Pyrazoles / Q000494:pharmacology*; D047069:Pyrazolones; D011725:Pyridines / Q000494:pharmacology*; D011728:Pyridones; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D027682:Cation Transport Proteins; D011720:Pyrazoles; D047069:Pyrazolones; D011725:Pyridines; D011728:Pyridones; D017382:Reactive Oxygen Species; C576694:setanaxib; D004317:Doxorubicin; D002118:Calcium
keywords: Ca(2+) transporters; Cardiotoxicity; Doxorubicin; GKT137831; NADPH oxidase; Reactive oxygen species