Purpose: To determine (1) the effect of a 40-minute steady-state run on muscle membrane integrity of elite athletes as reflected by serum creatine kinase (CK), (2) whether antioxidant supplementation (AS) with vitamins E and C has a protective effect, and (3) if a minimal blood concentration of vitamin E or C is required for any such protection. Methods: Fifteen elite-level endurance athletes ($\dot{\mathrm{V}}{\mathrm{O}}_2\text{max}=71.5\pm 1.2\mathrm{mL}·{\mathrm{kg}}^{-1}{\min }^{-1}$) were randomly assigned to 6 weeks AS (1000 IU·d⁻¹ natural vitamin E and 1000 mg·d⁻¹ vitamin C) or placebo. Using a double-blind crossover design and 4-week washout period, each treatment was followed by a 40-minute steady-state run at 3 mM blood lactate. Blood samples before and 0 and 24 hours after the run were assayed for serum and red cell α-tocopherol (α-TOH), serum ascorbate, and CK. Results: The AS produced a 2.5-fold, well-correlated (r = .84) increase in serum and red cell α-TOH (P < .001) that attenuated the increase in postrun CK (P = .01). There was no change in serum ascorbate with AS and no relationship with CK (P > .1). Curvilinear regression revealed some evidence that a critical level of serum α-TOH in the vicinity of 12 mg·L⁻¹ was required to attenuate CK efflux, a level only achieved with AS. Conclusion: The muscle membrane integrity of elite-level athletes is compromised even during steady-state running of moderate intensity and duration. The AS provided a protective effect, with evidence that a serum α-TOH concentration of around 12 mg·L⁻¹ is required.