title: Glutathione depletion in the guinea pig and its effect on the acute cochlear toxicity of ethacrynic acid.
abstract: There is controversy as to whether or not the acute cochlear toxicity of ethacrynic acid (EA) is dependent upon its metabolic conversion to EA-cysteine via conjugation with glutathione. In order to investigate this we examined the acute effects of EA on cochlear potentials in guinea pigs in which glutathione levels were decreased by prior administration of (+/-)-buthionine sulphoximine (BSO), an inhibitor of glutamylcysteine synthetase. First, we determined the effects of BSO on hepatic and renal glutathione levels in the guinea pig. Guinea pigs (pigmented animals of both sexes or male albino animals) were killed at intervals up to 72 hr after i.p. administration of 1.6 g kg-1 BSO. Livers, and also kidneys in the case of pigmented guinea pigs, were removed and total glutathione (GSH + GSSG) measured. Glutathione levels reached a nadir in the liver at 24-48 hr (11% of control) and in the kidneys at 24 hr (14% of control) after administration of BSO. Hepatic but not renal levels approached control values by 72 hr. There were no sex or strain differences. Pigmented guinea pigs were anaesthetised and their endocochlear potential and a.c. cochlear potential in response to a 4 kHz tone were measured using an intracochlear microelectrode. The depression of these potentials by i.v. administration of 60 mg kg-1 EA was not affected by administration of 1.6 g kg-1 BSO 24 hr earlier, despite profound depletion of glutathione. Also prior p.o. administration of N-acetyl-L-cysteine did not affect hepatic glutathione levels nor modify the toxicity of EA. These results suggest that the acute cochlear toxicity of EA is not altered by glutathione depletion, a finding which argues against a role for the metabolic activation of EA in its ototoxicity.
authors.affiliation: MRC Mechanisms of Drug Toxicity Research Group, Aston University, Birmingham, U.K.
authors.forename: C M
authors.initials: CM
authors.lastname: Lazenby
authors.forename: S J
authors.initials: SJ
authors.lastname: Lee
authors.forename: E S
authors.initials: ES
authors.lastname: Harpur
authors.forename: A
authors.initials: A
authors.lastname: Gescher
mesh_terms: D000818:Animals; D019328:Buthionine Sulfoximine; D003051:Cochlea / Q000187:drug effects*; D003545:Cysteine / Q000378:metabolism; D004976:Ethacrynic Acid / Q000378:metabolism / Q000633:toxicity*; D005260:Female; D005978:Glutathione / Q000032:analysis / Q000502:physiology*; D006168:Guinea Pigs; D007668:Kidney / Q000032:analysis; D008099:Liver / Q000032:analysis; D008297:Male; D008717:Methionine Sulfoximine / Q000031:analogs & derivatives / Q000494:pharmacology; D013045:Species Specificity
chemical_list: D008717:Methionine Sulfoximine; D019328:Buthionine Sulfoximine; D005978:Glutathione; D003545:Cysteine; D004976:Ethacrynic Acid