Endothelin stimulates vascular hydroxyl radical formation: effect of obesity

Reactive oxygen species (ROS) and endothelin-1 (ET-1) contribute to vascular pathophysiology in obesity. In this context, whether ET-1 modulates hydroxyl radical (OH) formation and the function of ROS/center dot OH in obesity is not known. In the present study, formation and function of ROS, including center dot OH, were investigated in the aorta of lean and leptin-deficient obese ob/ob mice. Hydroxyl radical formation was detected ex vivo using terephthalic acid in intact aortic rings and the involvement of ROS in ET-1-mediated vasoreactivity was analyzed using the antioxidant EPC-K1, a combination of alpha-tocopherol and ascorbic acid. Generation of either center dot OH, center dot O-2(-), and H2O2 was strongly inhibited by EPC-K1 (all P < 0.05). In obese mice, basal vascular center dot OH formation and ROS activity were reduced by 3-fold and 5-fold, respectively (P < 0.05 vs. lean). ET-1 markedly enhanced center dot OH formation in lean (6-fold, P < 0.05 vs. untreated) but not in obese mice. Obesity increased ET-1-induced contractions (P < 0.05 vs. lean), and ROS scavenging further enhanced the response (P < 0.05 vs. untreated). Exogenous ROS, including center dot OH caused stronger vasodilation in obese animals (P < 0.05 vs. lean), whereas endothelium-dependent relaxation was similar between lean and obese animals. In conclusion, we present a sensitive method allowing ex vivo measurement of vascular center dot OH generation and provide evidence that ET-1 regulates vascular center dot OH formation. The data indicate that in obesity, vascular formation of ROS, including center dot OH is lower, whereas the sensitivity to ROS is increased, suggesting a novel and important role of ROS, including center dot OH in the regulation of vascular tone in disease status associated with increased body weight.