2-Ethylhexyl diphenyl phosphate induces lung oxidative stress and pyroptosis in chicks

2-Ethylhexyl diphenyl phosphate (EHDPHP) is a widely used organophosphorus flame retardant and plasticizer easily released into the environment. Its biological toxicity is of great concern. The lung is considered a possible target organ for EHDPHP, but currently, there are limited studies on the biotoxicity of EHDPHP in poultry lungs. Therefore, the lungs were selected as the target organ to study the toxic effects of EHDPHP on chicks and their mechanisms of action. In this study, 7-day-old chicks were gavaged with different concentrations of EHDPHP, and lung samples were collected at 14, 28, and 42 days after intragastric administration. Lung histopathological and ultrapathological changes were examined by paraffin section-HE staining and transmission electron microscopy. The levels of lung damage markers (LDH) and oxidative stress markers (GSH-Px, SOD, and MDA) were detected by applying the kit. In contrast, lung cell pyroptosis-related factors (NLRP3, ASC, NF-κB, Pro-Caspase-1, IL-18, and IL-1β) and inflammatory factors (IL-6 and TNF-α) were assessed by using the qRT-PCR, Western blot and ELISA techniques. The results showed that EHDPHP induced pathological morphological changes and elevated LDH content in chick lungs, decreased lung antioxidant enzymes (GSH-Px and SOD) activities, increased peroxidation product MDA content and up-regulated the expression levels of cellular pyroptosis factors (NLRP3, ASC, NF-κB, Pro-Caspase-1, IL-18, and IL-1β), and the synthesis and secretion of inflammatory factors (IL-6 and TNF-α) were promoted. The above changes were EHDPHP dose-dependent. The results indicated that EHDPHP induced oxidative stress in chick lungs, resulting in oxidative damage to the lungs, and, intriguingly, the cellular pyroptosis pathway was activated, which was also involved in the process of EHDPHP-induced inflammatory damage in chick lungs. The results of this study revealed for the first time the damaging effects and mechanisms of EHDPHP on chick lungs. Also, they provided a scientific basis for further exploring the mechanisms of toxicity damage, safe use, and pollution control of EHDPHP.