An isolated rat heart model of intermittent hypoxia was used to investigate the impact of exogenous supplementation of glutathione and two thiol delivery vehicles on functional recovery during reoxygenation and whether efficacy was dependent on enhanced intracellular thiol concentration. Hearts from F344 rats were perfused in the Langendorff mode and exposed to three, 5 minute bouts of global, substrate free, normothermic hypoxia separated by 5 minute reoxygenation periods. Changes in coronary flow, heart rate, systolic and diastolic pressure, and rate pressure product were evaluated throughout in control hearts and compared with hearts in which one of the following was provided during the hypoxic periods: reduced glutathione (GSH, 1 or 10 mM), 10 mM GSH mono-ethyl ester (GSHMEE), or 1 mM L-2-oxothiozolidine-4-carboxylate (OZT). After three hypoxic periods plus reoxygenation, rate pressure product in control hearts was - 60% of pre-hypoxic values. Exposing hearts to 1 or 10 mM GSH, 10 mM GSHMEE, or 1 mM OZT significantly (p<0.05) enhanced post-hypoxic recovery of rate pressure product and attenuated the rise in diastolic pressure during hypoxia. This improvement in function was not associated with an elevated intracellular thiol concentration in treated hearts. Cumulative oxidative changes may occur during intermittent hypoxia via a mechanism localized on or near the sarcolemmal membrane. These changes appear to precede the appearance of significant intracellular oxidative stress and may be due to alterations in the reduced status of critical membrane bound proteins. Exogenously administered thiols attenuate protein alterations via a localized increase in thiol availability without an increase in gross measures of intracellular thiol or glutathione content.