Involvement of oxidative stress in the synthesis of metallothionein induced by mitochondrial inhibitors.

We previously reported that synthesis of metallothionein (MT) was induced by mitochondrial inhibitors such as 2,4-dinitrophenol (DNP) or antimycin A (Kondoh et al., 2001), which are potent inhibitors of mitochondrial respiration. Although the inhibitors are known to be radical generators in mitochondria, the involvement of oxidative stress in the synthesis of MT induced by mitochondrial inhibitors and the biological functions of MT remain obscure. In this study, therefore, we examined the involvement of oxidative stress in MT synthesis induced by mitochondrial inhibitors and the biological functions of MT. In cultured mouse fibroblast cells, the addition of DNP increased both MT concentration and MT mRNA level. Administration of DNP to L-buthionine-SR-sulfoximine (BSO)-pretreated mice increased hepatic lipid peroxidation and induction of MT synthesis. In addition, vitamin E prevented induction of MT synthesis as well as lipid peroxidation in the liver of mice caused by administration of DNP. Administration of mitochondrial inhibitor to mice elevated the levels of lipid peroxidation in the liver and mitochondria, and MT in the liver, indicating the generation of mitochondrial oxidative stress. These data suggest that the induction of MT synthesis by mitochondrial inhibitors is correlated with generation of oxidative stress in mitochondria. Furthermore, the level of DNP-induced alanine aminotransferase (ALT) activity, reflecting hepatic damage, was greater in MT-null mice than in wild-type mice, and intracellular accumulation of reactive oxygen species (ROS) caused by the action of mitochondrial inhibitors was greater in MT-null fibloblast cells than in wild-type cells. The results suggest that MT plays a role as a radical scavenger of intracellular ROS produced in mitochondria. Taken together, the results suggest that mitochondrial oxidative stress induces the synthesis of MT, which may contribute to regulation of mitochondrial ROS production.