We have previously observed that in vivo lipoic acid (LA) treatment induced a protective effect onto primary cortical neurons after brain injury. In an effort to better understand LA action mechanism in the brain, in the present study, we stressed brain cells in vitro and ex vivo and then analyzed by inmmunocytochemistry and biochemical assays, the changes induced by LA on cell survival and on the concentration of oxidative stress markers, such as glutathione (GSH), oxidized glutathione (GSSG), and malondialdehyde (MDA). The stressors used were lipopolysaccharide (LPS), dopamine, and l-buthionine-S,R-sulfoximine (BSO). Our results showed that LA decreased cell death and increased GSH/GSSG ratio in cells stressed by LPS + dopamine, suggesting that the mechanism underlying LA action is regeneration of GSSG to GSH. When cells were stressed by BSO, LA diminished cell death and decreased GSH/GSSG ratio. In this case, it could be concluded that, due to the low GSH basal levels, GSSG reduction is not possible and therefore it might be thought that cell death prevention might be mediated through other mechanisms. Finally, we induced chemical oxidative damage in brain homogenate. After LA treatment, GSH and GSH/GSSG ratio increased and MDA concentration decreased, demonstrating again that LA was not able to increase de novo GSH synthesis but is able to increase GSSG conversion to GSH. © 2016 American Chemical Society.