Impairment of diazoxide-induced formation of reactive oxygen species and loss of cardioprotection in connexin 43 deficient mice.
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 micromol/L) and valinomycin (10 nmol/L) induced similar ROS formation in wild-type (WT) and Cx43+/- cardiomyocytes. In contrast, diazoxide (200 micromol/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 microg/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.
Heinzel
Frank R
FR
Institut für Pathophysiologie, Universitätsklinikum Essen, Germany.
Luo
Yukun
Y
Li
Xiaokui
X
Boengler
Kerstin
K
Buechert
Astrid
A
García-Dorado
David
D
Di Lisa
Fabio
F
Schulz
Rainer
R
Heusch
Gerd
G
D000818:Animals; D018031:Connexin 43 / Q000502:physiology*; D003981:Diazoxide / Q000494:pharmacology*; D019157:Ischemic Preconditioning, Myocardial; D051379:Mice; D008929:Mitochondria, Heart / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000378:metabolism*; D020011:Protective Agents / Q000494:pharmacology; D017382:Reactive Oxygen Species; D014634:Valinomycin / Q000494:pharmacology; D024483:Vitamin K 3 / Q000494:pharmacology
D018031:Connexin 43; D020011:Protective Agents; D017382:Reactive Oxygen Species; D014634:Valinomycin; D024483:Vitamin K 3; D003981:Diazoxide