title: Biokinetics of chlorpromazine in primary rat and human hepatocytes and human HepaRG cells after repeated exposure.
abstract: Since drug induced liver injury is difficult to predict in animal models, more representative tests are needed to better evaluate these effects in humans. Existing in vitro systems hold great potential to detect hepatotoxicity of pharmaceuticals. In this study, the in vitro biokinetics of the model hepatotoxicant chlorpromazine (CPZ) were evaluated in three different liver cell systems after repeated exposure in order to incorporate repeated-dose testing into an in vitro assay. Primary rat and human hepatocytes, cultured in sandwich configuration and the human HepaRG cell line were treated daily with CPZ for 14 days. Samples were taken from medium, cells and well plastic at specific time points after the first and last exposure. The samples were analysed by HPLC-UV to determine the amount of CPZ in these samples. Based on cytotoxicity assays, the three models were tested at 1-2 μM CPZ, while the primary rat hepatocytes and the HepaRG cell line were in addition exposed to a higher concentration of 15-20 μM. Overall, the mass balance of CPZ decreased in the course of 24 h, indicating the metabolism of the compound within the cells. The largest decrease in parent compound was seen in the primary cultures; in the HepaRG cell cultures the mass balance only decreased to 50%. CPZ accumulated in the cells during the 14-day repeated exposure. Possible explanations for the accumulation of CPZ are a decrease in metabolism over time, inhibition of efflux transporters or binding to phospholipids. The biokinetics of CPZ differed between the three liver cell models and were influenced by specific cell properties as well as culture conditions. These results support the conclusion that in vitro biokinetics data are necessary to better interpret chemical-induced cytotoxicity data.
authors.affiliation: Institute for Risk Assessment Sciences, Utrecht University, Utrecht, The Netherlands. Electronic address: jessica.broeders@ctgb.nl.
authors.forename: Jessica J W
authors.initials: JJ
authors.lastname: Broeders
authors.affiliation: Kaly-Cell, 20A Rue du Général Leclerc, Plobsheim, France.
authors.forename: Céline
authors.initials: C
authors.lastname: Parmentier
authors.affiliation: Non-Clinical Safety, Merck Serono, Merck KGaA, Darmstadt, Germany; Institut für Angewandte Biowissenschaften, Karlsruhe Institute of Technology (KIT), Karlsruhe, Germany.
authors.forename: Germaine L
authors.initials: GL
authors.lastname: Truisi
authors.affiliation: Institut National de la Santé et de la Recherche Médicale U991, Université de Rennes 1, Rennes, France.
authors.forename: Rozenn
authors.initials: R
authors.lastname: Jossé
authors.affiliation: Kaly-Cell, 20A Rue du Général Leclerc, Plobsheim, France.
authors.forename: Eliane
authors.initials: E
authors.lastname: Alexandre
authors.affiliation: Institut National de la Santé et de la Recherche Médicale U991, Université de Rennes 1, Rennes, France.
authors.forename: Camille C
authors.initials: CC
authors.lastname: Savary
authors.affiliation: Non-Clinical Safety, Merck Serono, Merck KGaA, Darmstadt, Germany.
authors.forename: Philip G
authors.initials: PG
authors.lastname: Hewitt
authors.affiliation: Non-Clinical Safety, Merck Serono, Merck KGaA, Darmstadt, Germany; Institut für Angewandte Biowissenschaften, Karlsruhe Institute of Technology (KIT), Karlsruhe, Germany.
authors.forename: Stefan O
authors.initials: SO
authors.lastname: Mueller
authors.affiliation: Institut National de la Santé et de la Recherche Médicale U991, Université de Rennes 1, Rennes, France.
authors.forename: André
authors.initials: A
authors.lastname: Guillouzo
authors.affiliation: Kaly-Cell, 20A Rue du Général Leclerc, Plobsheim, France.
authors.forename: Lysiane
authors.initials: L
authors.lastname: Richert
authors.affiliation: National Institute of Public Health and the Environment (RIVM), Bilthoven, The Netherlands.
authors.forename: Jan C H
authors.initials: JC
authors.lastname: van Eijkeren
authors.affiliation: Institute for Risk Assessment Sciences, Utrecht University, Utrecht, The Netherlands.
authors.forename: Joop L M
authors.initials: JL
authors.lastname: Hermens
authors.affiliation: Institute for Risk Assessment Sciences, Utrecht University, Utrecht, The Netherlands.
authors.forename: Bas J
authors.initials: BJ
authors.lastname: Blaauboer
mesh_terms: D000818:Animals; D002460:Cell Line; D002746:Chlorpromazine / Q000008:administration & dosage / Q000493:pharmacokinetics*; D018492:Dopamine Antagonists / Q000008:administration & dosage / Q000493:pharmacokinetics*; D004305:Dose-Response Relationship, Drug; D004334:Drug Administration Schedule; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D051381:Rats
chemical_list: D018492:Dopamine Antagonists; D002746:Chlorpromazine
keywords: Biokinetics; Chlorpromazine; HepaRG cells; Primary hepatocytes