title: Evaluation of oxidative status in acetaminophen treated rat hepatocytes in culture.
abstract: The present study describes the estimation of acetaminophen (AAP) toxicity in cultured rat hepatocytes. We used different concentrations of AAP - 1, 2.5, 5, 10 and 20 mM, to test influence of AAP on cellular viability, functional capacity and oxidative status at given time intervals. WST 1 test showed decrease of dehydrogenase activity in 5, 10 and 20 mM AAP to 75 % of control values after 1 hour of incubation. At 12 h of treatment, all AAP concentrations decreased WST-1 signal; no enzyme activity was found since 18 h in cells treated with 20 mM AAP according to LDH leakage test performed at 24 h of incubation. Functional capacity was tested by albumin assay where the decrease was strictly related to AAP dose. Intracellular oxidative status was assessed by analysis of GSH/GSSG levels and time course of ROS production and glutathione reductase (GR) activity. Increased ROS production was found already after 3 h of incubation in 2.5, 5, 10 and 20 mM AAP, respectively. The highest ROS production was measured after 12 h treatment. GR activity was decreased already after 3 h of incubation and remained also decreased in cells treated with 2.5, 5, 10 and 20 mM AAP during further incubation.
authors.affiliation: Department of Physiology, Charles University in Prague, Hradec Králové, Czech Republic. Tomas.Rousar@upce.cz
authors.forename: T
authors.initials: T
authors.lastname: Rousar
authors.forename: O
authors.initials: O
authors.lastname: Kucera
authors.forename: P
authors.initials: P
authors.lastname: Křiváková
authors.forename: H
authors.initials: H
authors.lastname: Lotková
authors.forename: R
authors.initials: R
authors.lastname: Kanďár
authors.forename: V
authors.initials: V
authors.lastname: Mužáková
authors.forename: Z
authors.initials: Z
authors.lastname: Červinková
mesh_terms: D000082:Acetaminophen / Q000633:toxicity*; D000418:Albumins / Q000378:metabolism; D018712:Analgesics, Non-Narcotic / Q000633:toxicity*; D000818:Animals; D002478:Cells, Cultured; D005978:Glutathione / Q000378:metabolism; D019803:Glutathione Disulfide / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D008297:Male; D018384:Oxidative Stress / Q000187:drug effects*; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D000418:Albumins; D018712:Analgesics, Non-Narcotic; D017382:Reactive Oxygen Species; D000082:Acetaminophen; D005978:Glutathione; D019803:Glutathione Disulfide