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Cellular impact of combinations of endosulfan, atrazine, and chlorpyrifos on human primary hepatocytes and HepaRG cells after short and chronic exposures

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Abstract

Chronic exposure to low doses of pesticides present in the environment is increasingly suspected to cause major health issues to humans. Toxicological evaluations become more complex when the exposure concerns chemical combinations. Atrazine, chlorpyrifos, and endosulfan are pesticides used worldwide in agriculture and are therefore currently found at residual levels in food and the environment, even in countries in which they are now banned. Our study aimed to use Real-Time Cell Impedance Analyzer to investigate changes in phenotypical status of primary human hepatocytes and differentiated HepaRG cells induced by short and chronic exposures to these three chemicals. In contrast to the traditionally used endpoint cytotoxicity test, this technology allows kinetic measurements in real-time throughout the entire experiment. Our data show significantly higher cytotoxic effects of mixtures as compared to individual pesticides and a greater susceptibility of human hepatocytes as compared to HepaRG to short-term exposure (24 h). Repeated exposure over 2 weeks to endosulfan and endosulfan-containing mixture induced HepaRG cell death in a time- and dose-dependent manner. Of the typical genes involved in metabolism and cell-response to xenobiotics, we found an exposure time- and condition-dependent deregulation of the expression of CYP3A4 and UGT1A in HepaRG cells exposed to low doses of pesticides and mixtures. Our data demonstrate the usefulness of real-time cell monitoring in long-term toxicological evaluations of co-exposure to xenobiotics. In addition, they support but at the same time highlight certain limitations in the use of HepaRG cells as the gold standard liver cell model in toxicity studies.

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Fundings

The authors received a Public Institutional Funding from INRA, the French National Research Program 07-PNRA-006, and the ANSES (ES-021).

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Authors and Affiliations

  1. Institut National de la Recherche Agronomique (INRA), Laboratoire de Toxicologie Cellulaire et Moléculaire des Xénobiotiques, UMR 1331, 400 route des Chappes, BP 167, 06903, Sophia-Antipolis Cedex, France

    Ahmad Nawaz, Andrej Razpotnik, Georges de Sousa & Roger Rahmani

  2. UMR 1331 TOXALIM (Research Centre in Food Toxicology), Institut National de la Recherche Agronomique (INRA), 180 chemin de Tournefeuille, 31027, Toulouse, France

    Patrick Rouimi & Jean Pierre Cravedi

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  1. Ahmad Nawaz
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  2. Andrej Razpotnik
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  3. Patrick Rouimi
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  4. Georges de Sousa
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Correspondence to Georges de Sousa.

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Ahmad Nawaz and Andrej Razpotnik equally contributed to this work.

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Nawaz, A., Razpotnik, A., Rouimi, P. et al. Cellular impact of combinations of endosulfan, atrazine, and chlorpyrifos on human primary hepatocytes and HepaRG cells after short and chronic exposures. Cell Biol Toxicol 30, 17–29 (2014). https://doi.org/10.1007/s10565-013-9266-x

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