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Vol. 296, Issue 2, 642-649, February 2001
Department of Pharmacology and Toxicology, Medical College of
Wisconsin, Milwaukee, Wisconsin
Stimulation of the 
1-opioid receptor has been shown to
trigger ischemic preconditioning (IPC). Additionally, myocardial
ischemia/reperfusion induces the activation of extracellular
signal-regulated kinase (ERK). Therefore, we examined the role of ERK
in acute cardioprotection induced by 1-opioid receptor
stimulation or IPC. Infarct size (IS) was expressed as a percentage of
the area at risk (AAR). Control animals had an IS/AAR of 60.6 ± 1.8. IPC and 1-opioid receptor stimulation with TAN-67
reduced IS/AAR (8.2 ± 1.3 and 30.2 ± 2.4). Inhibition of
ERK with the selective MEK-1 antagonist, PD 098059 during IPC or TAN-67
administration significantly reduced cardioprotection (41.5 ± 6.4 and 63.0 ± 4.8). Western Blot analysis and subsequent
densitometry corroborated these observations. Control, TAN-67-, or
IPC-treated hearts were harvested after 0, 5, 15, and 30 min of
ischemia or 5, 30, and 60 min of reperfusion and separated into
cytosolic and nuclear fractions. Both isoforms of ERK (p44 and p42)
rapidly increased to greater levels throughout reperfusion in the
nuclear fraction of IPC- and opioid-treated versus control rats,
however, this increase was not attenuated by PD 098059. Conversely, the
rapid activation of the 44-kDa isoform of ERK after 5 min of
reperfusion in the cytosolic fraction was significantly increased in
IPC- and opioid-treated hearts versus control, and this increase was
abolished by pretreatment with PD 098059. Additionally, p42 was
activated in the cytosolic fraction of IPC-treated animals. These
results suggest a key role for the 44-kDa isoform of ERK in the
cytoplasm during cardioprotection induced by either IPC or stimulation
of the 1-opioid receptor.
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