Technical Report: Application of Rat Precision-cut Liver Slices for Toxicity Assessment In Vitro

The usefulness of precision-cut liver slices for toxicological evaluation in vitro was confirmed with a representative hepatotoxic agent acetaminophen. Liver slices obtained from normal and in vivo phenobarbital-treated rats were incubated with 0, 1.25, 2.5, 5, and 10 mM of acetaminophen for 24 hours. Intracellular potassium concentration dose-dependently decreased and lactate dehydrogenase (LDH) leakage increased in the slices from phenobarbital-treated rats, whereas no changes were detected in the slices from normal rats. The cytotoxicity of acetaminophen was also shown in the slices from normal rats when the slices were preincubated with phenobarbital in vitro for one hour. The acetaminophen cytotoxicity in the liver slices was enhanced by the preincubation with diethyl maleate, a depressant of hepatic glutathione, tributyltin acetate, an inhibitor of glutathione S-transferase, and 2, 6-dichloro-4-nitrophenol, an inhibitor of sulfotransferase. These data suggest that the formation and accumulation of the toxic metabolite are essential to induce the acetaminophen hepatotoxicity. Histopathological observation revealed centrilobular hepatocyte necrosis on the slices from phenobarbital-treated rats 24 hours after the exposure to acetaminophen. These results coincide with those reported in in vivo acetaminophen toxicity. The application of precision-cut liver slices is considered to be useful in toxicological evaluation of chemicals.