TBHQ alleviates pyroptosis and necroptosis in chicken alveolar epithelial cells induced by fine particulate matter from broiler houses

Fine particulate matter (PM(2.5)) from poultry houses has adverse effects on the health of animals and workers. Tert-butylhydroquinone (TBHQ), an antioxidant, is widely used in feed additives. The present study investigated the effects of TBHQ on broiler house PM(2.5)-induced damage in chicken primary alveolar epithelial cells (AECII) extracted from 16-day-old chicken embryos using the method of differential adhesion. AECII were exposed to PM(2.5) and TBHQ alone or in combination, and then, cell membrane integrity, pyroptosis, and necroptosis were detected. Our results showed that PM(2.5) from broiler houses caused cell rupture and loss of cell membrane integrity. This result was confirmed by the obvious increases in lactate dehydrogenase (LDH) release and propidium iodide (PI)-positive cells compared to the control group. In addition, the intracellular reactive oxygen species (ROS) levels and the expression levels of pyroptosis-related genes (NLRP3, IL-18, IL-1β) and necroptosis-related genes (RIPK3) were also significantly enhanced. However, TBHQ significantly inhibited intracellular ROS, improved cell viability, and reduced the release of LDH and the number of PI-positive cells compared to those in the PM(2.5) group. The expression levels of pyroptosis-related genes (Caspase-1, NLRP3, IL-18, IL-1β) and necroptosis-related genes (RIPK3) were also significantly decreased in the co-treatment group. In summary, these results indicated that TBHQ can alleviate PM(2.5)-mediated cell pyroptosis and necroptosis in chicken AECII and provide a basis for overcoming the danger that air pollutants from broiler houses pose to the health of chickens.