摘要：

To investigate the relationship between alterations of cytosolic Ca 2+ concentration and development of cytotoxicity, isolated rat hepatocytes were loaded with the fluorescent indicator Quin-2 AM and then incubated with non-toxic or toxic levels of menadione (2-methyl-1,4-napthoquinone) or tert -butyl hydroperoxide ( t -BH). The resulting changes in cytosolic Ca 2+ concentration were compared to those seen upon exposure of the hepatocytes to an α 1 -adrenergic agonist, phenylephrine, as well as to those induced by menadione and ( t -BH) in hepatocytes pretreated with agents that modify their toxicity. Exposure of hepatocytes to phenylephrine or non-toxic levels of menadione caused a moderate and transient increase in cytosolic Ca 2+ (0.7 μ M ) , whereas a toxic concentration of menadione produced a marked, sustained increase in Ca 2+ which fully saturated the binding capacity of Quin-2 (>1.5 μM). Treatment of the hepatocytes with the protective agent, dithiothreitol, prevented both the increase in cytosolic Ca 2+ and the cytotoxicity induced by menadione. On the other hand, pretreatment of cells with diethylmaleate to deplete intracellular glutathione made otherwise non-toxic concentrations of menadione cause both a sustained increase in cytosolic Ca 2+ and cytotoxicity. Similarly, toxic concentrations of t -BH also caused a sustained increase in cytosolic Ca 2+ . The iron chelator, desferrioxamine, and dithiothreitol (DTT), which protected the cells from t -BH toxicity, also prevented the sustained elevation of cytosolic Ca 2+ . Our findings provide further support for the hypothesis that a perturbation of intracellular Ca 2+ homeostasis is an early and critical event in the development of toxicity in hepatocytes exposed to oxidative stress.

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