ETHANOL INCREASES CYTOCHROME P4502E1 AND INDUCES OXIDATIVE STRESS IN ASTROCYTES

We demonstrate the presence of cytochrome P4502E1 (CYP2E1) in astrocyt es in primary culture, its induction by ethanol, and the concomitant g eneration of free radical species. Double immunofluorescence using ant i-CYP2E1 and anti-glial fibrillary acidic protein showed that CYP2E1 w as distributed over the cytoplasm and processes, although labeling was more pronounced over the nuclear membrane, Immunogold labeling confir med this pattern of distribution. Addition of 25 mM ethanol to the ast rocyte culture medium for 14 days resulted in an increase in the CYP2E 1 content, as determined by confocal microscopy and dot blot. In addit ion, ethanol induced a dose-dependent increase in the formation of rea ctive oxygen species that was partially prevented by incubating the as trocytes with anti-CYP2E1. Alcohol also induced a dose-dependent incre ase in malonaldehyde and hydroxynonenal formation and a depletion of t he glutathione (GSH) content, These results suggest that ethanol induc es oxidative damage in astrocytes, which could explain some of the tox ic effects of ethanol on these cells, such as cytoskeletal alterations , This assumption is supported here by the fact that an increase in GS H content prevents the deleterious effects of alcohol on the cytoskele ton of astrocytes, These results suggest the importance of oxidative s tress as a mechanism involved in alcohol-induced neural and brain dama ge.