Linking Oxidative Events to Inflammatory and Adaptive Gene Expression Induced by Exposure to an Organic Particulate Matter Component

Background: Toxicological studies have correlated inflammatory effects of diesel exhaust particles (DEP) with its organic constituents, such as the organic electrophile 1,2-naphthoquinone (1,2-NQ). Objective: To elucidate the mechanisms involved in 1,2-NQ–induced inflammatory responses, we examined the role of oxidant stress in 1,2-NQ–induced expression of inflammatory and adaptive genes in a human airway epithelial cell line. Methods: We measured cytosolic redox status and hydrogen peroxide (H(2)O(2)) in living cells using the genetically encoded green fluorescent protein (GFP)-based fluorescent indicators roGFP2 and HyPer, respectively. Expression of interleukin-8 (IL-8), cyclooxygenase-2 (COX-2), and heme oxygenase-1 (HO-1) mRNA was measured in BEAS-2B cells exposed to 1,2-NQ for 1–4 hr. Catalase overexpression and metabolic inhibitors were used to determine the role of redox changes and H(2)O(2) in 1,2-NQ–induced gene expression. Results: Cells expressing roGFP2 and HyPer showed a rapid loss of redox potential and an increase in H(2)O(2) of mitochondrial origin following exposure to 1,2-NQ. Overexpression of catalase diminished the H(2)O(2)-dependent signal but not the 1,2-NQ–induced loss of reducing potential. Catalase overexpression and inhibitors of mitochondrial respiration diminished elevations in IL-8 and COX-2 induced by exposure to 1,2-NQ, but potentiated HO-1 mRNA levels in BEAS cells. Conclusion: These data show that 1,2-NQ exposure induces mitochondrial production of H(2)O(2) that mediates the expression of inflammatory genes, but not the concurrent loss of reducing redox potential in BEAS cells. 1,2-NQ exposure also causes marked expression of HO-1 that appears to be enhanced by suppression of H(2)O(2). These findings shed light into the oxidant-dependent events that underlie cellular responses to environmental electrophiles.