Exploring the oxidative stress response mechanism triggered by environmental water samples
Environmental waters can contain a wide range of micropollutants. Bioanalytical test batteries using assays indicative of different stages of cellular toxicity pathways, such as adaptive stress responses, have been applied to a range of water samples. Oxidative stress response assays have proven to be sensitive tools, but the mechanism by which water samples are inducing the oxidative stress response remains unclear because both electrophiles and reactive oxygen species (ROS) may activate the Nrf2-antioxidant response element (ARE) pathway. The current study aimed to explore the underlying mechanisms of the oxidative stress response triggered by exposure to surface water extracts previously shown to be active in the ARE GeneBLAzer oxidative stress response assay. ROS formation and changes in glutathione (GSH) concentration were assessed in human liver cells exposed to water extracts from a large river in addition to individual chemicals that were detected in these water extracts and reported to be active in the ARE GeneBLAzer assay in a previous study. Many of the surface water samples induced ROS formation and decreased the GSH to glutathione disulfide (GSSG) ratio, suggesting that the formation of ROS is an important mechanism. However, some of the most responsive samples in the ARE GeneBLAzer assay, as well as the individual chemicals, did not have a significant effect on either ROS formation or the GSH/GSSG ratio, suggesting a different mechanism. Antioxidants can also induce the Nrf2-ARE pathway and the ARE GeneBLAzer assay may also detect compounds that activate ARE by Nrf2-independent mechanisms, thus further research is required to characterise active chemicals in oxidative stress response assays. However, these tests are still useful for quantifying the integrated cellular response to multiple molecular initiating events and can be used complementary to assays indicative of specific effects, such as receptor-mediated assays.