Effects of exercise training on vasodilatory protein expression and activity in rats

Abstract

Increased endothelium-dependent vasodilatation is associated with endurance exercise training. The purpose of this study was to test the hypothesis that increased endothelial nitric oxide synthase (eNOS) protein function, but not increased vascular smooth muscle sensitivity to NO, underlies augmented endothelium-dependent dilatation with training. To test these hypotheses, rats ran on a treadmill at 30 m/min (10% grade) for 60 min/day, 5 days/week, over 8–12 weeks (Trn). Training efficacy was demonstrated by greater (P < 0.05) hindlimb muscle citrate synthase activity and left ventricular mass–body mass ratio in Trn compared with sedentary control rats (Sed). Expression of eNOS protein in the aorta was increased with training (Sed, 1.00 ± 0.18 normalized units; Trn, 1.55 ± 0.23; P < 0.05). Aortic NOS activity was, however, unchanged by training (Sed, 1,505 ± 288 fmol/h/mg protein; Trn, 1,650 ± 247; n.s.). Expression of heat shock protein 90 and protein kinase B/Akt was not different between groups, nor was their association with eNOS. In follow-up series of rats, phosphorylated eNOS content (Serine 1177) was similar for Sed and Trn in both the aorta and gastrocnemius feed artery. Aortic NOS activity with eNOS phosphorylation status preserved was also similar between groups. Finally, cGMP concentration with a NO donor did not differ between groups (Sed, 73.0 ± 20.2 pmol/mg protein; Trn, 62.5 ± 12.9; n.s.). These findings indicate that training-induced increases in eNOS protein expression are not coupled to augmented function, illustrating the complexity of eNOS regulation. Further, they show that vascular sensitivity to NO is not altered by exercise training.