Published online before print August 11, 2005, doi: 10.1161/01.RES.0000181171.65293.65
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© 2005 American Heart Association, Inc.
Integrative Physiology |
Impairment of Diazoxide-Induced Formation of Reactive Oxygen Species and Loss of Cardioprotection in Connexin 43 Deficient Mice
From the Institut für Pathophysiologie (F.R.H., Y.L., X.L., K.B., A.B., R.S., G.H.), Universitätsklinikum Essen, Germany; Servicio de Cardiologia (D.G.-D.), Hospital Vall d’Hebron, Barcelona, Spain; and the Dipartimento di Chimica Biologica (F.D.L.), Universita di Padova, Italy.
Correspondence to Prof. Dr. med. Dr. h. c. Gerd Heusch, Direktor des Instituts für Pathophysiologie, Zentrum für Innere Medizin, Universitätsklinikum Essen, Hufelandstr. 55, 45122 Essen, Germany. E-mail gerd.heusch{at}uni-essen.de
Protection by ischemic preconditioning is lost in cardiomyocytes and hearts of heterozygous connexin 43 deficient (Cx43+/–) mice. Because connexin 43 (Cx43) is localized in cardiomyocyte mitochondria and mitochondrial Cx43 content is increased with ischemic preconditioning, we now tried to identify a functional defect at the level of the mitochondria in Cx43+/– mice by use of diazoxide and menadione. Diazoxide stimulates the mitochondrial formation of reactive oxygen species (ROS) and menadione generates superoxide at multiple intracellular sites; both substances elicit cardioprotection through increased ROS formation. ROS formation in response to the potassium ionophore valinomycin was also measured for comparison. Menadione (2 µmol/L) and valinomycin (10 nmol/L) induced similar ROS formation in wild-type (WT) and Cx43+/– cardiomyocytes. In contrast, diazoxide (200 µmol/L) increased ROS formation by 43±10% versus vehicle in WT, but only by 18±4% in Cx43+/– cardiomyoctes (P<0.05). Two hour–simulated ischemia and oxygenated, hypo-osmolar reperfusion reduced viability as compared with normoxia (WT: 7±1% versus 39±2%, Cx43+/–: 8±1% versus 40±3%, P<0.01). Although menadione protected WT and Cx43+/– cardiomyocytes, diazoxide increased viability (17±2%, P<0.01) in WT, but not in Cx43+/– (9±1%). Menadione (37 µg/kg i.v.) before 30 minutes coronary occlusion and 2 hour reperfusion reduced infarct size in WT and Cx43+/– mice (24±4% versus 24±5%). In contrast, diazoxide (5 mg/kg i.v.) reduced infarct size in WT (35±4% versus 55±3% of area at risk, P<0.01), but not in Cx43+/– mice (56±2% versus 54±3%). Cardiomyocytes of Cx43+/– mice have a specific functional deficit in ROS formation in response to diazoxide and accordingly less protection.
Key Words: ischemia • signal transduction
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