Biokinetics of chlorpromazine in primary rat and human hepatocytes and human HepaRG cells after repeated exposure. Issue 1 (25th December 2015)

• Record Type: Journal Article
• Authors: Broeders, Jessica J.W.; Parmentier, Céline; Truisi, Germaine L.; Jossé, Rozenn; Alexandre, Eliane; Savary, Camille C.; Hewitt, Philip G.; Mueller, Stefan O.; Guillouzo, André; Richert, Lysiane; van Eijkeren, Jan C.H.; Hermens, Joop L.M.; Blaauboer, Bas J.
• Is Part Of: Toxicology in vitro. Volume 30:Issue 1 Part A (2015)
• Page Start: 52
• Page End: 61
• Publication Date: 2015-12-25
• Subjects: BSA bovine serum albumin -- CPZ chlorpromazine -- CYPs cytochrome P450 enzymes -- DMEM Dulbecco's modified eagle medium -- DMSO dimethyl sulfoxide -- EDTA ethylenediaminetetra acetic acid -- HPLC high pressure liquid chromatography -- ITS insulin–transferrin–selenium -- PBS phosphate buffered saline -- TC10 10% toxic concentration; Biokinetics -- Primary hepatocytes -- HepaRG cells -- Chlorpromazine; Toxicity testing -- In vitro -- Periodicals; Toxicology -- Periodicals; 615.9
• DOI: 10.1016/j.tiv.2014.08.012
• Languages: English
• ISSNs: 0887-2333

Abstract

Highlights: Chlorpromazine accumulated in the cells during the repeated exposure. Differences in biokinetics were found in the different liver in vitro systems. The in vitro biokinetics were dependent on specific cell properties and culture conditions. 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.