Alterations of skeletal muscle redox state via antioxidant supplementation have the potential to impact contractile function and vascular smooth muscle tone. The effects of antioxidants on the regulation of muscle O2 delivery–O2 utilization () matching (which sets the microvascular partial pressure of O2; ) in young healthy muscle are not known. Therefore, the purpose of this study was to test the effects of acute antioxidant supplementation on rat spinotrapezius muscle force production, blood flow, and (phosphorescence quenching). Anaesthetized male Fischer 344 × Brown Norway rats (6–8 months old) had their right spinotrapezius muscles either exposed for measurement of blood flow and (n= 13) or exteriorized for measurement of muscle force production (n= 6). Electrically stimulated 1 Hz twitch contractions (∼7–9 V) were elicited for 180 s, and measurements were made before and after acute intra-arterial antioxidant supplementation (76 mg kg−1 ascorbic acid, 52 mg kg−1 tempol) dissolved in saline and infused over 30 min. The principal effects of antioxidants were a ∼25% decrease (P < 0.05) in contracting spinotrapezius muscle force production concurrent with reductions in muscle blood flow and at rest and during contractions (P < 0.05 for both). Antioxidant supplementation reduced the resting baseline (before, 29.9 ± 1.2 mmHg; after, 25.6 ± 1.3 mmHg; P < 0.05), and this magnitude of depression was sustained throughout the rest-to-exercise transition (steady-state value before, 16.4 ± 0.7 mmHg; after, 13.6 ± 0.9 mmHg; P < 0.05). In addition, the time constant of the decrease was reduced after antioxidant supplementation (before, 23.4 ± 4.3 s; after, 15.6 ± 2.7 s; P < 0.05). These results demonstrate that antioxidant supplementation significantly impacts the control of in young rats at rest and during contractions.