A correlation between glutathione levels and cellular damage in isolated hepatocytes.

To obtain high levels of glutathione in isolated hepatocytes an isolation procedure shorter than 16 min was used. This procedure gave a moderately high yield of viable cells (200--300 X 10(6) cells/10 g liver) with 44 +/- 3 nmol of glutathione/10(6) cells. Incubation in Krebs-Henseleit solution containing 2% albumin resulted in a continuous loss of reduced glutathione from the cells, while incubation in a medium containing amino acids and horse serum resulted in increased levels, suggesting active synthesis for 5 h. A short and apparently harmless depletion of reduced glutathione was induced by diethylmaleate or cumene hydroperoxide. A depletion of reduced glutathione lasting more than 1 h was accompanied by an increased cellular leakage. The depletion was induced by either diethylmaleate plus paracetamol or diethylmaleate alone in higher concentrations. A common mechanism for these toxic responses is suggested.