2-Ethylhexyl salicylate induces developmental toxicity through oxidative stress: Insights from in silico prediction, Drosophila experiments and AOP framework.

2-Ethylhexyl salicylate (EHS), a prevalent organic ultraviolet filter in personal-care and industrial goods, enters humans and ecosystems via dermal contact and environmental discharge, triggering worries over its long-term developmental toxicity. This study integrated network toxicology, Drosophila melanogaster experiments, dose-dependent transcriptomics, and the adverse outcome pathway (AOP) framework to investigate the mechanisms of EHS-induced developmental toxicity. Network toxicology yielded 168 candidate targets and 11 hub genes, whose enrichment pointed to oxidative‑stress‑related processes and the PI3K/AKT cascade. Molecular docking confirmed stable EHS-hub-protein binding. Parental EHS exposure significantly reduced pupal number and eclosion rate in offspring, indicating developmental toxicity. Additionally, exposure of EHS to adult Drosophila revealed decreases in body weight, triglyceride levels, superoxide dismutase activity, and climbing ability, accompanied by increased catalase activity and malondialdehyde content. N-acetylcysteine rescue experiments further validated the pivotal role of oxidative stress in EHS-induced toxicity. Based on dose-dependent transcriptomic profiling, an AOP was constructed with increased reactive oxygen species (ROS) as the molecular initiating event, progressing through lipid peroxidation, mitochondrial dysfunction, and DNA damage, ultimately leading to developmental toxicity. MAPK and PI3K/AKT signaling pathways might play critical roles in this process. This study uncovers EHS developmental-toxicity mechanisms and supports its risk evaluation.