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Department of Anesthesiology, University of North Carolina, Chapel Hill, North Carolina 27599
The aims of this study were to
determine whether preconditioning blocks cardiocyte apoptosis
and to determine the role of mitochondrial ATP-sensitive K+
(KATP) channels and the protein kinase C 
-isoform
(PKC-) in this effect. Ventricular myocytes from 10-day-old chick
embryos were used. In the control series, 10 h of simulated
ischemia followed by 12 h of reoxygenation resulted in
42 ± 3% apoptosis (n = 8). These results
were consistent with DNA laddering and TdT-mediated dUTP nick-end
labeling (TUNEL) assay. Preconditioning, elicited with three cycles of
1 min of ischemia separated by 5 min of reoxygenation before
subjection to prolonged simulated ischemia, markedly attenuated the apoptotic process (28 ± 4%, n = 8). The
selective mitochondrial KATP channel opener diazoxide (400 µmol/l), given before ischemia, mimicked preconditioning effects to
prevent apoptosis (22 ± 4%, n = 6). Pretreatment with 5-hydroxydecanoate (100 µmol/l), a
selective mitochondrial KATP channel blocker, abolished
preconditioning (42 ± 2%, n = 6). In addition,
the effects of preconditioning and diazoxide were blocked with the
specific PKC inhibitors Gö-6976 (0.1 µmol/l) or chelerythrine
(4 µmol/l), given at simulated ischemia and reoxygenation.
Furthermore, preconditioning and diazoxide selectively activated
PKC- in the particulate fraction before simulated ischemia
without effect on the total fraction, cytosolic fraction, and PKC
-isoform. The specific PKC activator phorbol 12-myristate 13-acetate
(0.2 µmol/l), added during simulated ischemia and
reoxygenation, mimicked preconditioning to block apoptosis. Opening mitochondrial KATP channels blocks cardiocyte
apoptosis via activating PKC- in cultured ventricular
myocytes. Through this signal transduction, preconditioning blocks
apoptosis and preserves cardiac function in
ischemia-reperfusion.
protein kinase C -isoform; mitochondrial ATP-sensitive potassium
channels
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