Extracellular redox status regulates Nrf2 activation through mitochondrial reactive oxygen species.

The redox status of the extracellular compartment has only just been elucidated as a mechanism controlling intracellular signal transduction and correlates with aging, diabetes, heart disease and lung fibrosis. In the present paper, we describe a mechanism by which oxidizing extracellular environments, as maintained by the cysteine/cystine (Cys/CySS) redox couple, induce mitochondria-derived ROS (reactive oxygen species) generation and cause the activation of Nrf2 (nuclear factor-erythroid 2-related factor 2), inducing an antioxidant response. NIH 3T3 cells were cultured in medium with extracellular Cys/CySS redox potentials (Eh), ranging from 0 to -150 mV. Cellular and mitochondrial ROS production significantly increased in cells incubated under more oxidizing extracellular conditions (0 and -46 mV). Trx2 (thioredoxin-2) is a mitochondrial-specific oxidoreductase and antioxidant and became oxidized in cells incubated at 0 or -46 mV. MEFs (mouse embryonic fibroblasts) from Trx2-overexpressing transgenic (Trx2 Tg) mice produced less intracellular ROS compared with WT (wild-type) MEFs at the more oxidizing extracellular conditions. Nrf2 activity was increased in WT MEFs at the 0 or -46 mV conditions, but was inhibited in Trx2 Tg MEFs under the same conditions. Furthermore, Nrf2-regulated gene expression was significantly increased in the WT MEFs, but not in the Trx2 Tg MEFs. These results show that the Cys/CySS redox status in the extracellular compartment regulates intracellular ROS generated primarily in the mitochondria, which play an important role in the activation of Nrf2 and up-regulation of antioxidant and detoxification systems.