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Articles in PresS, published online ahead of print September 19, 2002
Am J Physiol Heart Circ Physiol, 10.1152/ajpheart.00717.2002
Submitted on August 16, 2002
Accepted on September 12, 2002
1 Heart Foundation Research Centre, Griffith University Gold Cost Campus, Southport, QLD, Australia
* To whom correspondence should be addressed. E-mail: j.headrick{at}mailbox.gu.edu.au.
Relative roles of mitochondrial (mito) KATP channels, protein kinase C (PKC), and adenosine kinase (AK) in adenosine-mediated protection were assessed in Langendorff perfused mouse hearts subjected to 20 min ischemia and 45 min reperfusion. Control hearts recovered 72±3 mmHg ventricular pressure (50% pre-ischemia), and released 23±2 IU/g lactate dehydrogenase (LDH). Adenosine (50 µM) during ischemia-reperfusion improved recovery (149±8 mmHg) and reduced LDH efflux (5±1 IU/g). Treatment during ischemia alone was less effective. Treatment with 50 µM diazoxide (mito KATP opener) during ischemia and reperfusion enhanced recovery, and was equally effective during ischemia alone. A3 agonism (100 nM 2-chloro-N6-(3-iodobenzyl)-adenosine-5-N-methyluronamide), A1 agonism (N6-cyclohexyladenosine), and AK inhibition (10 µM iodotubercidin) all reduced necrosis to the same extent as adenosine, but less effectively reduced contractile dysfunction. These responses were abolished by 100 µM 5-hydroxydecanoate (5-HD, mito KATP channel blocker) or 3 µM chelerythrine (PKC inhibitor). However, protective effects of adenosine during ischemia-reperfusion were resistant to 5-HD and chelerythrine, and only abolished when inhibitors were co-infused with iodotubercidin. Data indicate adenosine-mediated protection via A1/A3ARs is mito KATP channel and PKC dependent, with evidence for a downstream location of PKC. Adenosine provides additional and substantial protection via phosphorylation to 5'-AMP, primarily during reperfusion.
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