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1 Medicine/Cardiovascular Diease, University of Alabama at Birmingham, Birmingham, Alabama, United States
2 Physiology & Biophysics, University of Alabama at Birmingham, United States
3 Cell Biology, University of Alabama at Birmingham, Birmingham, Alabama, United States
* To whom correspondence should be addressed. E-mail: jchatham{at}uab.edu.
We have shown that in the perfused heart glucosamine improved functional recovery following ischemia and this appeared to be mediated via an increase in O-Linked N-acetylglucosamine (O-GlcNAc) levels on nucleocytoplasmic proteins. Several kinase pathways, specifically Akt and the mitogen activated protein kinases (MAPK) p38 and ERK1/2, which have been implicated in ischemic cardioprotection, have been reported to be modified in response to increased O-GlcNAc levels. Therefore, the goals of this study were to determine the effect of ischemia on O-GlcNAc levels and to evaluate whether the cardioprotection resulting from glucosamine treatment could be attributed to changes in ERK1/2, Akt and p38 phosphorylation. Isolated rat hearts were perfused with or without 5mM glucosamine and were subjected to 5, 10 or 30 min low flow ischemia (LFI) or 30 min LFI and 60min reperfusion. Glucosamine treatment attenuated ischemic contracture and improved functional recovery at the end of reperfusion. Glucosamine treatment increased flux through the hexosamine biosynthesis pathway and increased O-GlcNAc levels but had no effect on ATP levels. Glucosamine did not alter the response of either ERK1/2 or Akt to ischemia/reperfusion; however it significantly attenuated the ischemia-induced increase in p38 phosphorylation and paradoxically increased p38 phosphorylation at the end of reperfusion. These data support the notion that O-GlcNAc may play an important role as an internal stress response and that glucosamine-induced cardioprotection may be mediated via the p38 MAPK pathway.
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