Mitochondrial ATP-dependent potassium channels: novel effectors of cardioprotection?
BACKGROUND
Brief interruptions of coronary blood flow paradoxically protect the heart from subsequent prolonged ischemia. The basis of such endogenous cardioprotection, known as "ischemic preconditioning," remains uncertain. Pharmacological evidence has implicated ATP-dependent potassium (KATP) channels in the mechanism of preconditioning; however, the effects of sarcolemmal KATP channels on excitability cannot account for the protection.


RESULTS
We simultaneously measured flavoprotein fluorescence, an index of mitochondrial redox state, and sarcolemmal KATP currents in intact rabbit ventricular myocytes. Our results show that diazoxide, a KATP channel opener, selectively activates mitochondrial KATP channels. Diazoxide induced reversible oxidation of flavoproteins with an EC50 of 27 micromol/L but did not activate sarcolemmal KATP channels. The subcellular site of diazoxide action is further localized to mitochondria by confocal imaging of fluorescence arising from flavoproteins and tetramethylrhodamine ethyl ester. In a cellular model of simulated ischemia, inclusion of diazoxide decreased the rate of cell death to about half of that in controls. Both the redox changes and protection are inhibited by the KATP channel blocker 5-hydroxydecanoic acid.


CONCLUSIONS
Our results demonstrate that diazoxide targets mitochondrial but not sarcolemmal KATP channels and imply that mitochondrial KATP channels may mediate the protection from KATP channel openers.
Liu
Y
Y
Department of Medicine, Johns Hopkins University, Baltimore, MD 21205, USA.
Sato
T
T
O'Rourke
B
B
Marban
E
E
D000255:Adenosine Triphosphate; D000818:Animals; D002478:Cells, Cultured; D003981:Diazoxide / Q000494:pharmacology*; D005420:Flavoproteins / Q000187:drug effects / Q000378:metabolism; D005453:Fluorescence; D006352:Heart Ventricles / Q000166:cytology / Q000187:drug effects; D019157:Ischemic Preconditioning, Myocardial; D018613:Microscopy, Confocal; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D017202:Myocardial Ischemia / Q000378:metabolism; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D010084:Oxidation-Reduction; D015221:Potassium Channels / Q000187:drug effects* / Q000378:metabolism; D011817:Rabbits; D012508:Sarcolemma / Q000187:drug effects* / Q000378:metabolism
D005420:Flavoproteins; D015221:Potassium Channels; D000255:Adenosine Triphosphate; D003981:Diazoxide