AMP 579, an
adenosine A(1)/A(2) receptor agonist, has a strong anti-
infarct effect when administered just before reperfusion. Because oxidative stress has been proposed to contribute to
myocardial reperfusion injury, we tested whether
AMP 579 can reduce the production of reactive
oxidant species (ROS) during reoxygenation in cultured chick embryonic cardiomyocytes. The intracellular
fluorescent probe 2',7'-dichlorofluorescin diacetate (DCFH) was used to detect ROS. The cells were subjected to 60 min of simulated
ischemia, followed by either 15 min or 3 h of reoxygenation.
AMP 579 (0.5 and 1 microM), when started 10 min before reoxygenation, significantly reduced ROS generation from 4.86 +/- 0.30 (arbitrary units) in untreated cells to 2.72 +/- 0.31 and 1.85 +/- 0.14, respectively (P < 0.05). Cell death that was assessed by
propidium iodide uptake was markedly reduced by
AMP 579 (49.6 +/- 4.7% of control cells vs. 25.4 +/- 2.4%, P < 0.05). In contrast,
adenosine did not alter ROS generation or cell death. Attenuation of ROS production by
AMP 579 was completely prevented by simultaneous exposure of cells to the selective
adenosine A(2) antagonist 8-(13-chlorostyryl)
caffeine. These results indicate that
AMP 579 directly protects cardiomyocytes from
reperfusion injury by a mechanism that attenuates intracellular
oxidant stress. Furthermore,
adenosine could not duplicate these effects.