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Articles in PresS, published online ahead of print May 16, 2002
Am J Physiol Heart Circ Physiol, 10.1152/ajpheart.00173.2002
Submitted on February 28, 2002
Accepted on May 8, 2002
1 Molecular and Cellular Physiology, Louisiana State University Health Sciences Center, Shreveport, LA, USA; First Department of Internal Medicine, Kyoto Prefectural University of Medicine, Kyoto, Japan
2 Molecular and Cellular Physiology, Louisiana State University Health Sciences Center, Shreveport, LA, USA
3 First Department of Internal Medicine, Kyoto Prefectural University of Medicine, Kyoto, Japan
* To whom correspondence should be addressed. E-mail: rkorth{at}lsuhsc.edu.
Long-term ethanol consumption at low to moderate levels exerts cardioprotective effects in the setting of ischemia and reperfusion (I/R). The aims of this study were to determine whether: (1) a single orally administered dose of ethanol (ethanol preconditioning or EtOH-PC) would induce a biphasic temporal pattern of protection (early and late phases) against the inflammatory responses to I/R and (2) adenosine and nitric oxide (NO) act as initiators of the late phase of protection. Ethanol was administered as a bolus to C57BL/6 mice at a dose that achieved a peak plasma concentration of ~45 mg/dl 30 min after gavage and returned to control levels within 60 of alcohol ingestion. The superior mesenteric artery was occluded for 45 min followed by 60 min of reperfusion beginning 10 min or 1, 2, 3, 4, or 24 hrs after ethanol ingestion and the numbers of fluorescently-labeled rolling and firmly adherent (stationary) leukocytes in single postcapillary venules of the small intestine were quantified using intravital microscopic approaches. I/R induced marked increases in leukocyte rolling and adhesion, effects that were attenuated by EtOH-PC 2-3 hr prior to I/R (early phase), absent when assessed after 10 min, 1 hr, and 4 hr of ethanol ingestion, with an even more powerful late phase of protection re-emerging when I/R was induced 24 hr later. The anti-inflammatory effects of late EtOH-PC were abolished by treatment with adenosine deaminase, an adenosine A2 (but not A1) receptor antagonist, or an NO synthase (NOS) inhibitor, during the period of EtOH-PC. Preconditioning with an adenosine A2 (but not an A1) receptor agonist in lieu of ethanol 24 hr prior to I/R mimicked the protective actions of late phase EtOH-PC. Like EtOH-PC, the effect of preconditioning with an adenosine A2 receptor agonist was abrogated by coincident NOS inhibition. These findings suggest that EtOH-PC induces a biphasic temporal pattern of protection against the proinflammatory effects of I/R. In addition, our observations are consistent with the hypothesis that the late phase of EtOH-PC is triggered by nitric oxide formed secondary to adenosine A2-receptor-dependent activation of nitric oxide synthase during the period of ethanol exposure.
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