Endothelial vasomotor function decreases with increasing age. Extracellular superoxide dismutase (ecSOD) protects against vascular dysfunction in several disease states. The purpose of this study was to determine whether endogenous ecSOD protects against endothelial dysfunction in old mice. Vasomotor function of the aorta was studied ex vivo in wild-type (ecSOD(+/+)) and ecSOD-deficient (ecSOD(-/-)) mice at 11 (adult) and 29 (old) mo of age. Maximal relaxation to acetylcholine (10(-4) M) was impaired in vessels from adult ecSOD(-/-) mice [75 +/- 3% (mean +/- SE)] compared with wild-type mice (89 +/- 2%, P < 0.05). Maximal relaxation to acetylcholine (10(-4) M) was profoundly impaired in aorta from old ecSOD(-/-) mice (45 +/- 5%) compared with wild-type mice (75 +/- 4%, P < 0.05). There was a significant correlation between expression of ecSOD and maximal relaxation to acetylcholine in adult and old mice. Tempol (1 mM), a scavenger of superoxide, improved relaxation in response to acetylcholine (63 +/- 8%) in old ecSOD(-/-) mice (P < 0.05), but not wild-type mice (75 +/- 4%). Maximal relaxation to sodium nitroprusside was similar in aorta from adult and old wild-type and ecSOD(-/-) mice. Quantitative RT-PCR showed a decrease in mRNA levels of ecSOD and catalase in aorta of old mice and an increase in levels of TNFalpha and Nox-4 in aorta of old mice compared with adult mice. The findings support the hypothesis that impaired antioxidant mechanisms may contribute to cumulative increases in oxidative stress and impaired endothelial function in old mice. In conclusion, endogenous ecSOD plays an important role in protection against endothelial dysfunction during aging.