Oxidative stress impairs endothelium-dependent dilation (EDD) with aging in healthy sedentary adults. Increased cytochrome P-450 2C9 (CYP 2C9) signaling can contribute to oxidative stress-mediated suppression of EDD, but its role in aging is unknown. We hypothesized that inhibition of CYP 2C9 signaling with sulfaphenazole would improve EDD in older, but not young, healthy sedentary adults. At baseline, increases in forearm blood flow (FBF; venous occlusion plethysmography) in response to brachial artery infusions of ACh (1, 2, 4, and 8 microg.100 ml forearm volume(-1).min(-1)), an endothelium-dependent dilator, were smaller in older [n = 14, 63 +/- 1 (SE) yr] than in young (n = 11, 23 +/- 2 yr) adults (P < 0.05), with a reduction in peak FBF of 32% (11.8 +/- 1.7 vs. 17.3 +/- 2.3 ml.100 ml tissue(-1).min(-1)). Infusion of sulfaphenazole at doses that block CYP 2C9 signaling in humans did not affect the FBF responses to ACh in the older (peak FBF = 13.0 +/- 4.3 ml.100 ml tissue(-1).min(-1), P = 0.41) or the young (peak FBF = 17.1 +/- 1.9 ml.100 ml tissue(-1).min(-1), P = 0.55) adults. Coadministration of the nitric oxide inhibitor l-NMMA and sulfaphenazole decreased the FBF response to ACh in young and older subjects (P < 0.05); the effect was smaller in the older subjects, but group differences in EDD remained (P < 0.05). Endothelium-independent dilation assessed with sodium nitroprusside was not different in the young and older subjects. These results provide the first support for the concept that increased CYP 2C9 signaling does not contribute to impairments in EDD with aging in healthy adults.