Role of Nrf2 in preventing oxidative stress induced chloride current alteration in human lung cells

The lung tissue is one of the main targets of oxidative stress due to external sources and respiratory activity. In our previous work, we have demonstrated in that O₃ exposure alters the Cl^- current-voltage relationship, with the appearance of a large outward rectifier component mainly sustained by outward rectifier chloride channels (ORCCs) in human lung epithelial cells (A549 line). In the present study, we have performed patch clamp experiments, in order to identify which one of the O₃ byproducts (4hydroxynonenal (HNE) and/or H₂ O₂ ) was responsible for chloride current change. While 4HNE exposition (up to 25 μM for 30' before electrophysiological analysis) did not reproduce O₃ effect, H₂ O₂ produced by glucose oxidase 10 mU for 24 hr before electrophysiological analysis mimicked O₃ response. This result was confirmed treating the cell with catalase (CAT) before O₃ exposure (1,000 U/ml for 2 hr): CAT was able to rescue Cl^- current alteration. Since CAT is regulated by Nrf2 transcription factor, we pre-treated the cells with the Nrf2 activators, resveratrol and tBHQ. Immunochemical and immunocytochemical results showed Nrf2 activation with both substances that lead to prevent OS effect on Cl^- current. These data bring new insights into the mechanisms involved in OS-induced lung tissue damage, pointing out the role of H₂ O₂ in chloride current alteration and the ability of Nfr2 activation in preventing this effect.