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J Biol Chem, Vol. 273, Issue 29, 18092-18098, July 17, 1998
From the Department of Medicine, University of Chicago,
Chicago, Illinois 60637
Reactive oxygen species (ROS) have been proposed
to participate in the induction of cardiac preconditioning. However,
their source and mechanism of induction are unclear. We tested whether brief hypoxia induces preconditioning by augmenting mitochondrial generation of ROS in chick cardiomyocytes. Cells were preconditioned with 10 min of hypoxia, followed by 1 h of simulated ischemia and
3 h of reperfusion. Preconditioning decreased cell death from 47 ± 3% to 14 ± 2%. Return of contraction was observed in
3/3 preconditioned versus 0/6 non-preconditioned
experiments. During induction, ROS oxidation of the probe
dichlorofluorescin (sensitive to H2O2)
increased ~2.5-fold. As a substitute for hypoxia, the addition of
H2O2 (15 µmol/liter) during normoxia also
induced preconditioning-like protection. Conversely, the ROS signal
during hypoxia was attenuated with the thiol reductant
2-mercaptopropionyl glycine, the cytosolic Cu,Zn-superoxide dismutase
inhibitor diethyldithiocarbamic acid, and the anion channel inhibitor
4,4'-diisothiocyanato-stilbene-2,2'-disulfonate, all of which also
abrogated protection. ROS generation during hypoxia was attenuated by
myxothiazol, but not by diphenyleneiodonium or the nitric-oxide
synthase inhibitor L-nitroarginine. We conclude that
hypoxia increases mitochondrial superoxide generation which initiates preconditioning protection. Furthermore, mitochondrial anion channels and cytosolic dismutation to
H2O2 may be important steps for oxidant
induction of hypoxic preconditioning.
Reactive Oxygen Species Released from Mitochondria during Brief
Hypoxia Induce Preconditioning in Cardiomyocytes
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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