title: Ouabain protects rat hearts against ischemia-reperfusion injury via pathway involving src kinase, mitoKATP, and ROS.
abstract: We showed recently that mitochondrial ATP-dependent K(+) channel (mitoK(ATP)) opening is required for the inotropic response to ouabain. Because mitoK(ATP) opening is also required for most forms of cardioprotection, we investigated whether exposure to ouabain was cardioprotective. We also began to map the signaling pathways linking ouabain binding to Na(+)-K(+)-ATPase with the opening of mitoK(ATP). In Langendorff-perfused rat hearts, 10-80 microM ouabain given before the onset of ischemia resulted in cardioprotection against ischemia-reperfusion injury, as documented by an improved recovery of contractile function and a reduction of infarct size. In skinned cardiac fibers, a ouabain-induced protection of mitochondrial outer membrane integrity, adenine nucleotide compartmentation, and energy transfer efficiency was evidenced by a decreased release of cytochrome c and preserved half-saturation constant of respiration for ADP and adenine nucleotide translocase-mitochondrial creatine kinase coupling, respectively. Ouabain-induced positive inotropy was dose dependent over the range studied, whereas ouabain-induced cardioprotection was maximal at the lowest dose tested. Compared with bradykinin (BK)-induced preconditioning, ouabain was equally efficient. However, the two ligands clearly diverge in the intracellular steps leading to mitoK(ATP) opening from their respective receptors. Thus BK-induced cardioprotection was blocked by inhibitors of cGMP-dependent protein kinase (PKG) or guanylyl cyclase (GC), whereas ouabain-induced protection was not blocked by either agent. Interestingly, however, ouabain-induced inotropy appears to require PKG and GC. Thus 5-hydroxydecanoate (a selective mitoK(ATP) inhibitor), N-(2-mercaptopropionyl)glycine (MPG; a reactive oxygen species scavenger), ODQ (a GC inhibitor), PP2 (a src kinase inhibitor), and KT-5823 (a PKG inhibitor) abolished preconditioning by BK and blocked the inotropic response to ouabain. However, only PP2, 5-HD, and MPG blocked ouabain-induced cardioprotection.
authors.affiliation: Institut National de la Santé et de la Recherche, Médicale U441, University Victor Segalen-Bordeaux 2, University Hospital of Bordeaux, Bordeaux, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Pasdois
authors.forename: Casey L
authors.initials: CL
authors.lastname: Quinlan
authors.forename: Abraham
authors.initials: A
authors.lastname: Rissa
authors.forename: Liliane
authors.initials: L
authors.lastname: Tariosse
authors.forename: Béatrice
authors.initials: B
authors.lastname: Vinassa
authors.forename: Alexandre D T
authors.initials: AD
authors.lastname: Costa
authors.forename: Sandrine V
authors.initials: SV
authors.lastname: Pierre
authors.forename: Pierre
authors.initials: P
authors.lastname: Dos Santos
authors.forename: Keith D
authors.initials: KD
authors.lastname: Garlid
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D000818:Animals; D002316:Cardiotonic Agents / Q000627:therapeutic use; D003401:Creatine / Q000378:metabolism; D004195:Disease Models, Animal; D006339:Heart Rate / Q000187:drug effects; D008297:Male; D008929:Mitochondria, Heart / Q000187:drug effects / Q000378:metabolism; D051336:Mitochondrial Membranes / Q000502:physiology; D015428:Myocardial Reperfusion Injury / Q000517:prevention & control*; D010042:Ouabain / Q000627:therapeutic use*; D010101:Oxygen Consumption; D010539:Permeability; D015221:Potassium Channels / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism*; D019061:src-Family Kinases / Q000378:metabolism*
chemical_list: D002316:Cardiotonic Agents; D015221:Potassium Channels; D017382:Reactive Oxygen Species; C117780:mitochondrial K(ATP) channel; D010042:Ouabain; D000244:Adenosine Diphosphate; D019061:src-Family Kinases; D003401:Creatine