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J. Biol. Chem., Vol. 276, Issue 28, 26357-26364, July 13, 2001
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From the Oxidative stress can cause significant cell death
by apoptosis. We performed studies in L-cells to explore whether prior
exposure to oxidative stress ("oxidative preconditioning") can
protect the cell against the apoptotic consequences of subsequent
oxidative insults and to establish the mediators in the preconditioning signaling cascade. Cells were preconditioned with three 5-min exposures
to H2O2, followed by 10-h recovery and
subsequent exposure to 600 µM
H2O2 for 10 h. A single 10-h exposure to
H2O2 induced substantial apoptotic cell death
(~90%), as determined by enzyme-linked immunosorbent assay, TUNEL
(terminal deoxyribonucleotide transferase-mediated dUTP nick end
labeling), and Annexin V methods, but apoptosis was largely prevented
in preconditioned cells. The degree of cytoprotection depended on
the strength of preconditioning or H2O2
concentration (20~600 µM). Transient increases in
mitogen-activated protein kinase (MAPK), p38, and JNK/SAPK
activities and sustained protein kinase B (Akt) activation,
accompanied by drastically reduced caspase 3 activity, were seen after
preconditioning. The expression levels of these kinases were unaltered.
Inhibitors of p38 (SB203580) and phosphoinositide 3-kinase (PI3K,
LY294002) pathways abolished the protection provided by
preconditioning. We conclude that oxidative preconditioning protects
cells against apoptosis and that this effect involves MAPK and PI3K/Akt
pathways. This system may be important in regulating apoptotic cell
death in development and disease states.
Oxidative Preconditioning and Apoptosis in L-cells
ROLES OF PROTEIN KINASE B AND MITOGEN-ACTIVATED PROTEIN
KINASES*
§,¶,,
**, and
Research Center, Montreal Heart Institute,
Montreal, Quebec H1T 1C8, the Department of Medicine, University
of Montreal, Montreal, Quebec H3C 3J7, and the ** Department of
Pharmacology and Therapeutics, McGill University, Montreal, Quebec
H3A 2T5, Canada
*
This work was supported in part by the Canadian Institute of
Health Research, the Heart and Stroke Foundation of Quebec, and the
Fonds de la Recherche de l'Institut de Cardiologie de Montreal (to
Z. W.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
A research scholar of the Heart and Stroke Foundation of
Canada. To whom correspondence should be addressed: Research Center, Montreal Heart Institute, 5000 Belanger East, Montreal, Quebec H1T 1C8,
Canada. Tel.: 514-376-3330; Fax: 514-376-1355; E-mail: wangz@icm.
umontreal.ca.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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