Development of intracellular
calcium overload is an important pathophysiological factor in
myocardial ischemia/reperfusion or
anoxia/reoxygenation injury. Recent studies have shown that
Sodium Ferulate (SF) stimulates
nitric oxide (NO) production and exerts a cardioprotective effect in the
ischemia-reperfused heart. However, it has not been determined whether the cardioprotection of SF is associated with suppression of Ca(2+) overload via NO/
cyclic GMP (cGMP)/
cGMP-dependent protein kinase (PKG) pathway. In this work, after cardiomyocytes were incubated with 100, 200, 400, or 800 microM SF for 3 h,
anoxia/reoxygenation injury was induced and intracellular Ca(2+) concentration,
NO synthase (NOS) activity,
guanylate cyclase activity, NO, and cGMP formation were measured appropriately. The results showed that treatment with SF concentration-dependently inhibited
calcium overload induced by
anoxia/reoxygenation. We also demonstrated that SF (100-800 microM) concentration dependently enhanced NO and cGMP formation through increasing NOS activity and
guanylate cyclase activity in the cardiomyocytes. On the contrary, inhibition of
calcium overload by SF was markedly attenuated by addition of an NOS inhibitor, an NO scavenger, an
soluble guanylate cyclase inhibitor, and a PKG inhibitor:
N(G)-nitro-l-arginine methyl ester (
L-NAME, 100 microM), 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazole-1-oxyl-3-oxide (c-
PTIO, 1.0 microM), 1H-[1, 2, 4] oxadiazolo [4, 3-alpha] quinoxalin-1-one (ODQ, 20 microM) and
KT5823 (0.2 microM), respectively. Our findings indicate that SF significantly attenuates
anoxia/reoxygenation-induced Ca(2+) overload and improves cell survival in cultured cardiomyocytes through NO/cGMP/PKG signal pathway.