title: A nonfeminizing estrogen analog protects against ethanol withdrawal toxicity in immortalized hippocampal cells.
abstract: We have shown that 17beta-estradiol protects against ethanol withdrawal toxicity in rats. Here, we investigated whether a cellular model of ethanol withdrawal could be developed in a cultured hippocampal cell line (HT22) and whether an adamantyl-containing nonfeminizing estrogen analog, ZYC26 [(3-hydroxy-2-adamantyl(1)-4-methyl-estra-1,3,5(10)-17-one], protects against ethanol withdrawal toxicity. HT22 cells were exposed to ethanol (0-500 mM) for 24 h in the presence or absence of ZYC26 or 17beta-estradiol. The ethanol solution was then removed from the cells for 4 h to create ethanol withdrawal. Samples were collected at the end of a 24-h ethanol exposure or at 4 h of ethanol withdrawal to assess cell viability using a calcein assay, lipid peroxidation by measuring malondialdehyde, and protein oxidation by measuring carbonyl contents. When tested, ethanol concentrations were constantly maintained during a 24-h ethanol exposure and eliminated at 4 h of ethanol withdrawal. Ethanol withdrawal decreased cell viability and increased the levels of malondialdehyde and carbonyls more than ethanol exposure. ZYC26 reduced the cell death and malondialdehyde levels at a lower dose (1 microM) than 17beta-estradiol (10 microM). The increased carbonyl contents were reduced only by ZYC26 treatment. These data suggest that ethanol withdrawal can be created in HT22 cells in a manner that is more toxic than ethanol exposure and that ZYC26 is a more potent cytoprotectant than 17beta-estradiol against cell death and oxidative damage induced by ethanol withdrawal. Therefore, ZYC26 can be a potential alternative estrogen therapy for a cellular and oxidative imbalance associated with ethanol withdrawal.
authors.affiliation: Department of Pharmacology and Neuroscience, University of North Texas Health Science Center at Fort Worth, 3500 Camp Bowie Boulevard, Fort Worth, TX 76107, USA. mjung@hsc.unt.edu
authors.forename: Marianna E
authors.initials: ME
authors.lastname: Jung
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Wilson
authors.forename: James W
authors.initials: JW
authors.lastname: Simpkins
mesh_terms: D000218:Adamantane / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D019610:Cytoprotection; D004967:Estrogens / Q000494:pharmacology*; D004970:Estrone / Q000031:analogs & derivatives* / Q000494:pharmacology; D000431:Ethanol / Q000633:toxicity*; D006624:Hippocampus / Q000166:cytology / Q000187:drug effects*; D015227:Lipid Peroxidation / Q000187:drug effects; D008315:Malondialdehyde / Q000032:analysis; D050050:Protein Carbonylation; D013375:Substance Withdrawal Syndrome / Q000378:metabolism* / Q000473:pathology
chemical_list: C515153:3-hydroxy-2-adamantyl(1)-4-methyl-estra-1,3,5(10)-17-one; D004967:Estrogens; D004970:Estrone; D000431:Ethanol; D008315:Malondialdehyde; D000218:Adamantane