EFFECTS OF REACTIVE OXYGEN METABOLITES ON NOREPINEPHRINE-INDUCED VASOCONSTRICTION

Aortic rings, 4 mm in length, were obtained from rats and placed on isometric force transducers in oxygenated Krebs buffer. Following a period of stabilization, the cumulative dose response relationship to norepinephrine was assessed. The vessels were washed and allowed to return to baseline in Krebs buffer containing xanthine (0.5 mM). Xanthine oxidase (0.1 U/ml) was then added to the bath and vessels incubated for 30 min. The vessels were resuspended in Krebs buffer and cumulative dose-response curves to norepinephrine reevaluated. The results indicate that generation of reactive oxygen metabolites by xanthine/xanthine oxidase decreases the pD(2) from 7.80 +/- 0.04 to 7.40 +/- 0.09 with the endothelium intact. Removal of the endothelium did not attenuate the contractile dysfunction, indicating that endothelial-derived metabolites were not mediating the loss of vasoconstrictor effectiveness. Maximal tension development did not differ between normal and oxidized vessel rings. Introduction of oxypurinol (0.2 mg/ml) to the bath prevented the loss of constrictor responsiveness, thereby confirming that all of the oxidants were derived from the xanthine/xanthine oxidase reaction. Superoxide dismutase (200 U/ml) partially prevented the loss of norepinephrine responsiveness produced by xanthine oxidase-derived radicals. The pD(2) in the SOD + xanthine/xanthine oxidase-treated vessels rings (7.19 +/- 0.11) was significantly lower than control vessel rings (7.49 +/- 0.04) and significantly higher than the xanthine/xanthine oxidase-treated vessels (6.89 +/- 0.06). Catalase (1000 U/ml) also partially attenuated the loss of vascular norepinephrine responsiveness. The pD(2) for the catalase + xanthine/xanthine oxidase-treated vessels (7.15 +/- 0.02) was significantly lower than control vessels (7.39 +/- 0.07) and significantly higher than the xanthine/xanthine oxidase-treated vessels (6.82 +/- 0.11). The pD(2) of vessels treated with a combination of SOD and catalase (7.40 +/- 0.10) did not differ from control vessels (7.49 +/- 0.12). The results of this study indicate that reactive oxygen species produced by the interaction of xanthine with xanthine oxidase depress norepinephrine-induced vasoconstriction. The loss of vasoconstrictor responsiveness appears to involve both superoxide and hydrogen peroxide.