The link between
endothelial nitric oxide synthase (eNOS) activation and vascular diameter during
ischemia-reperfusion was investigated in the rat heart. After short (<30 min) and long (>45 min) time of
ischemia conferred by coronary artery occlusion of the rats, reperfusion caused dilatation and constriction of arterioles, respectively. Partial
oxygen pressure (pO2) measurement of the heart by the
electrode confirmed the hyper-perfusion and no-reflow phenomena during reperfusion, as well as
myocardial ischemia. The vascular diameter was correlated with phosphorylation of Akt and
serine 1177 residue of eNOS, and formation of NO-bound
guanylate cyclase (GC) by immuoflorescence study. Western blotting confirmed the phosphorylation of eNOS-Ser1177 depending on
ischemia time. The constriction during reperfusion after 45 min of
ischemia is supposedly caused by the inhibition of Akt-mediated eNOS-Ser1177 phosphorylation, which was suppressed by a PKC inhibitor
chelerythrine, or ROS scavengers N-2-mercaptopropionyl
glycine (MPG) and 4,5-Dihydroxy-1, 3-benzenedisulfonic
acid disodium
salt (
Tiron). However, an
endothelin receptor antagonist BQ123 alleviated the vasoconstriction by increasing NO availability but not eNOS-Ser1177 phosphorylation. Thus, vascular patency is correlated with eNOS-Ser1177 phosphorylation in association with ROS, and PKC during reperfusion.
Endothelin inhibits vasodilatation by reducing NO availability during reperfusion.