Circulation Research. 2001;88:1267-1275
Published online before print June 7, 2001, doi: 10.1161/hh1201.092094
Published online before print June 7, 2001, doi: 10.1161/hh1201.092094
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© 2001 American Heart Association, Inc.
Cellular Biology |
Mitochondrial ATP-Sensitive Potassium Channels Inhibit Apoptosis Induced by Oxidative Stress in Cardiac Cells
From the Institute of Molecular Cardiobiology, The Johns Hopkins University, Baltimore, Md. Current address of A.O. is Georg August Universität Göttingen, Abteilung Kardiologie und Pneumologie, Göttingen, Germany.
Correspondence to Eduardo Marbán, MD, PhD, Institute of Molecular Cardiobiology, The Johns Hopkins University, 720 Rutland Ave, 844 Ross Bldg, Baltimore, MD 21205. E-mail marban{at}jhmi.edu
Abstract
Abstract—Mitochondria
can either enhance or suppress cell death. Cytochrome
c release from mitochondria and
depolarization of the mitochondrial membrane potential (
) are
crucial events in triggering apoptosis. In contrast, activation
of mitochondrial ATP-sensitive potassium
(mitoKATP) channels prevents lethal
ischemic injury in vivo, implicating these channels as key
players in the process of ischemic preconditioning. We probed
the relationship between mitoKATP channels and
apoptosis in cultured neonatal rat cardiac
ventricular myocytes. Incubation with 200 µmol/L hydrogen
peroxide induced TUNEL positivity, cytochrome
c translocation, caspase-3
activation, poly(ADP-ribose) polymerase cleavage, and dissipation of
. Pharmacological opening of mitoKATP
channels by diazoxide (100 µmol/L) preserved mitochondrial integrity
and suppressed all markers of apoptosis. Diazoxide prevented
depolarization in a concentration-dependent manner
(EC50 
40 µmol/L, with saturation by 100
µmol/L), as shown by both flow cytometry and quantitative image
analysis of cells stained with fluorescent
indicators. These cytoprotective effects of diazoxide were reproduced
by pinacidil, another mitoKATP agonist, and
blocked by the mitoKATP channel
antagonist 5-hydroxydecanoate (500 µmol/L). Our findings
identify a novel mitochondrial pathway that is protective against
apoptosis. The results also pinpoint
mitoKATP channels as logical therapeutic targets
in diseases of enhanced apoptosis and oxidative
stress.
Key Words: apoptosis • ischemia • oxidative stress
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