Induction of extracellular ATP mediates increase in intracellular thioredoxin in RAW264.7 cells exposed to low-dose -rays

Ohshima, Yasuhiro; Kitami, Akihiro*; Kawano, Ayumi*; Tsukimoto, Mitsutoshi*; Kojima, Shuji*

Extracellular ATP, which is released in response to various stresses, regulates the expression of intracellular antioxidants through activation of P2 receptors. We have recently found that low-dose -rays induce ATP release from the exposed cells. However, it is not yet clear whether or not the radiation-induced extracellular ATP modulates the cellular redox balance. Here, we investigated whether or not -ray irradiation-induced release of extracellular ATP contributes to the induction of cellular antioxidant thioredoxin-1 (Trx-1), using mouse macrophage-like RAW264.7 cells. Irradiation with -rays or exogenously added ATP increased expression of Trx-1, and in both cases the increase was blocked by pretreatment with an ecto-nucleotidase, apyrase. Then, involvement of ATP-dependent reactive oxygen species (ROS) generation in the increase of antioxidant capacity was examined. ATP stimulation promoted generation of intracellular ROS and also increased Trx-1 expression. The increase of Trx-1 expression was significantly suppressed by pre-treatment of the cells with antioxidants. In conclusion, the -ray irradiation-induced release of extracellular ATP may, at least in part, contribute to production of ROS via purinergic signaling, leading to promotion of intracellular antioxidants as an adaptive response to an oxidative stress.