Antiapoptotic effect of nicorandil mediated by mitochondrial atp-sensitive potassium channels in cultured cardiac myocytes.
OBJECTIVE
We examined whether nicorandil, a clinically useful drug for the treatment of ischemic syndromes, inhibits myocardial apoptosis.


BACKGROUND
Nicorandil has been reported to have a cardioprotective action through activation of mitochondrial ATP-sensitive potassium (mitoK(ATP)) channels. Based on our recent observation that mitoK(ATP) channel activation has a remarkable antiapoptotic effect in cultured cardiac cells, we hypothesized that the protective effects of nicorandil may be at least partially due to an antiapoptotic effect.


METHODS
Cultured neonatal rat cardiac myocytes were exposed to hydrogen peroxide to induce apoptosis. Effects of nicorandil were evaluated using a number of apoptotic markers.


RESULTS
Exposure to 100 microM hydrogen peroxide resulted in apoptotic cell death as shown by TUNEL positivity, cytochrome c translocation, caspase-3 activation and dissipation of mitochondrial inner membrane potential (Delta(Psi)(m)). Nicorandil (100 microM) suppressed all of these markers of apoptosis. Notably, nicorandil prevented Delta(Psi)(m) depolarization in a concentration-dependent manner (EC(50) approximately 40 microM, with saturation by 100 microM), as shown by fluorescence-activated cell sorter analysis of cells stained with a fluorescent Delta(Psi)(m)-indicator, tetramethylrhodamine ethyl ester (TMRE). Time-lapse confocal microscopy of individual cells loaded with TMRE shows that nicorandil suppresses Delta(Psi)(m) loss. Subcellular calcein localization revealed inhibition of the mitochondrial permeability transition by nicorandil. These protective effects of nicorandil were blocked by the mitoK(ATP) channel antagonist 5-hydroxydecanoate.


CONCLUSIONS
Our findings identify nicorandil as an inhibitor of apoptosis induced by oxidative stress in cardiac myocytes, and confirm the critical role of mitoK(ATP) channels in inhibiting apoptosis.
Akao
Masaharu
M
Institute of Molecular Cardiobiology, The Johns Hopkins University, Baltimore, Maryland 21205, USA.
Teshima
Yasushi
Y
Marbán
Eduardo
E
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002317:Cardiovascular Agents / Q000494:pharmacology*; D002478:Cells, Cultured; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects; D008929:Mitochondria, Heart / Q000187:drug effects* / Q000502:physiology; D009206:Myocardium / Q000166:cytology; D020108:Nicorandil / Q000494:pharmacology*; D018384:Oxidative Stress / Q000187:drug effects; D015221:Potassium Channels / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley
D002317:Cardiovascular Agents; D015221:Potassium Channels; D020108:Nicorandil; D000255:Adenosine Triphosphate; D006861:Hydrogen Peroxide