Although the
adenosine A(3) receptor agonist N(6)-(3-iodobenzyl)-adenosine-5'-N-methylcarboxamide (
IB-MECA) has been reported to be cardioprotective at reperfusion, little is known about the mechanisms underlying the protection. We hypothesized that
IB-MECA may protect the heart at reperfusion by preventing the opening of
mitochondrial permeability transition pore (mPTP) through inactivation of
glycogen synthase kinase (
GSK) 3beta.
IB-MECA (1 microM) applied during reperfusion reduced
infarct size in isolated rat hearts, an effect that was abrogated by the selective A3 receptor antagonist 1,4-dihydro-2-methyl-6-phenyl-4-(phenylethynyl)-3,5-pyridinedicarboxylic
acid 3-ethyl-5-[(3-nitrophenyl)-methyl]
ester (MRS1334) (100 nM). The effect of
IB-MECA was abrogated by the
mPTP opener
atractyloside (20 microM), implying that the action of
IB-MECA may be mediated by inhibition of the
mPTP opening. In cardiomyocytes,
IB-MECA attenuated
oxidant-induced loss of mitochondrial membrane potential (DeltaPsim), which was reversed by MRS1334.
IB-MECA also reduced Ca2+-induced mitochondrial swelling.
IB-MECA enhanced phosphorylation of
GSK-3beta (Ser9) upon reperfusion, and the
GSK-3 inhibitor
3-(2,4-dichlorophenyl)-4-(1-methyl-1H-indol-3-yl)-1H-pyrrole-2,5-dione (
SB216763) (3 microM) mimicked the protective effect of
IB-MECA by attenuating both
infarction and the loss of DeltaPsim. In addition, the effect of
IB-MECA on
GSK-3beta was reversed by
wortmannin (100 nM), and
IB-MECA was shown to enhance Akt phosphorylation upon reperfusion. In contrast,
rapamycin (2 nM) failed to affect
GSK-3beta phosphorylation by
IB-MECA, and
IB-MECA did not alter phosphorylation of either mTOR (Ser2448) or 70s6K (Thr389). Taken together, these data suggest that
IB-MECA prevents
myocardial reperfusion injury by inhibiting the
mPTP opening through the inactivation of
GSK-3beta at reperfusion.
IB-MECA-induced
GSK-3beta inhibition is mediated by the
PI3-kinase/Akt signal pathway but not by the mTOR/
p70s6K pathway.