Tacrine-induced Reactive Oxygen Species in a Human Liver Cell Line: The Role of Anethole Dithiolethione as a Scavenger

Toxicology in Vitro : an International Journal Published in Association with BIBRA
R A OsseniJ-M Warnet

Abstract

The mechanisms leading to tacrine (THA) hepatotoxic effects are not yet fully understood. Reactive oxygen species (ROS) overproduction and intracellular reduced glutathione (GSH) depletion are common mechanisms involved in drug toxicity. The aim of this study was to investigate, on the human liver cell line HepG2, whether THA at human blood concentrations induces ROS production stimulation and/or GSH depletion. A possible effect of a free radical scavenger, anethole dithiolethione (ADT), was also assessed. ROS production was measured with a fluorogen probe 2',7'-dichlorofluorescin diacetate (DCFH-DA). Reduced GSH and cell viability were measured with, respectively, monochlorobimane (mBCl) and neutral red probes. Assays were performed directly on living adherent cells in 96-well microplates, and sensitive fluorescent detection used microplate cytofluorimetry with cold light fluorimetry technology. The results showed that THA induced a concentration-dependent increase in ROS production and a decrease in GSH. Furthermore, for THA concentrations between 10 and 100 mum, ADT protected cells from ROS production stimulation and GSH depletion induced by THA. In conclusion, our in vitro study demonstrates that oxidative stress, evidenced...Continue Reading

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