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Articles in PresS, published online ahead of print January 17, 2002
Am J Physiol Heart Circ Physiol, 10.1152/ajpheart.00760.2001
Submitted on August 30, 2001
Accepted on January 16, 2002
1 Thoracic and Cardiovascular Surgery, Kansai Medical University, Moriguchi, Japan
2 Cardio-Thoracic Surgery, Beijing Friendship Hospital, Beijing, China
* To whom correspondence should be addressed. E-mail: otanih{at}takii.kmu.ac.jp.
Although adenosine is an important mediator of ischemic preconditioning (IPC), its relative contribution to IPC remains unknown. Because adenosine is formed through the hydrolysis of ATP, the present study investigated the role of ATP and adenosine in IPC. Isolated and buffer perfused rat heart underwent IPC by 3 cycles of 5 minutes ischemia and 5 min reperfusion before 25 minutes of global ischemia. Rate pressure products (RPP) 30 minutes after reperfusion was taken as an end-point of functional protection. Interstitial fluid (ISF) adenine nucleotides and adenosine were measured by a cardiac microdialysis technique. Inhibition of IPC-induced recovery of RPP was partial by the adenosine receptor antagonist 8-(p-sulfophenyl)theophylline (SPT; 100 µM) or by the structurally distinct P2Y purinoceptor antagonists suramin (300 µM) or reactive blue (RB; 10 µM), but was additive when SPT was given with suramin or RB. A P2X antagonist pyridoxal-phosphate-6-azophenyl-2',4'-disulphonic acid tetrasodium (50 µM) had no effect on functional protection. The improved functional recovery was not significantly affected by an ecto-5'-nucleotidase inhibitor, 
, ß-methylene adenosine diphosphate (AMP-CP; 100 µM) alone but was inhibited by AMP-CP plus SPT, suramin or RB. ISF ATP and adenosine increased temporarily by 10-fold during IPC. AMP-CP augmented the increase in ISF ATP associated with the decrease in ISF adenosine. There was a reciprocal correlation between the ISF concentration of ATP and adenosine in preconditioned hearts. In addition, there was significant correlation between ISF adenosine and ATP and the inhibitory potency of SPT and suramin or RB against functional protection conferred by IPC. These results suggest that extracellular ATP and adenosine play a complementary role in IPC through P2Y purinoceptors and adenosine receptors, respectively.
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