INTRACELLULAR GLUTATHIONE IN HUMAN HEPATOCYTES INCUBATED WITH S-ADENOSYL-L-METHIONINE AND GSH-DEPLETING DRUGS

The present study was undertaken to investigate (a) whether S-adenosyl-L-methionine (SAMe) added to culture medium can increase intracellular glutathione (GSH) levels in human hepatocytes and (b) whether SAMe can prevent the GSH depletion found in human hepatocytes incubated with GSH-depleting drugs (paracetamol, opiates, ethanol). Incubation of hepatocytes with increasing concentrations of SAMe resulted in a dose-dependent elevation of intracellular GSH content, which reached its maximum (35% increase) at 30-mu-M after 20 h. SAMe, as the only sulfur source in the medium, was efficient in repleting GSH-depleted hepatocytes following treatment with diethyl maleate. Incubation of human hepatocytes with SAMe attenuated the GSH depletion of cells incubated with toxic concentrations of paracetamol (2 mM), heroin (0.5 mM) and methadone (0.2 mM). A decrease in GSH due to exposure of hepatocytes to 50 mM ethanol was prevented when SAMe was simultaneously added to ethanol, and human hepatocytes maintained their GSH levels like non ethanol-treated cells. The experimental results of our work give the first direct evidence of the ability of exogenously administered SAMe to increase intracellular GSH levels in human hepatocytes and to prevent the GSH depletion caused by paracetamol, opiates and ethanol.