2-Chloroacetophenone is an effective glutathione depletor in isolated rat hepatocytes

The glutathione (GSH) depleting effect of 2-chloroacetophenone (CN) was studied in freshly isolated rat hepatocytes. CN proved to be more effective in depleting GSH than diethylmaleate, phorone or styrene oxide. The reaction between GSH and CN followed a 1:1 stoichiometry, allowing adjustment of cellular GSH concentrations at distinct levels. After incubating cells (8 mg protein/ml) with 200 mumol CN/l for 5 min, GSH depletion was almost complete without signs of cytotoxicity. At 300 mumol/l CN, GSH depletion persisted, and cytotoxicity occurred after 30 min. Activities of cytochrome P450 dependent enzymes, even at concentrations up to 500 mumol CN/l, were only marginally affected. Therefore, CN is of particular value for in vitro studies at decreased availability of GSH.