EPR imaging has been used to visualize redox status in oxidative brain diseases, but the role of cerebral glutathione (GSH) is not clear. In this study, using the mouse model of GSH depletion with diethylmaleate (DEM), the role of GSH in brain redox status was examined. The remarkable change in redox status in DEM-treated mouse brain was visualized with EPR imaging, and in vitro assay showed decrease in the level of GSH. ME-MRI clearly visualized blood-brain barrier dysfunction in DEM-treated mice. Results indicate that GSH plays an important role in the maintenance of both brain redox status and BBB integrity.