Porcine epidemic diarrhea virus induces PANoptosis in piglet intestinal cells via Z-RNA/ZBP1/ROS pathway-mediated oxidative stress activation.

Abstract

Porcine epidemic diarrhea virus (PEDV) is a type of coronavirus that infects pigs, resulting in high mortality rates in piglets and posing a significant threat to the swine industry. However, the biological mechanisms underlying PEDV-induced intestinal damage and the role of oxidative stress in this context remain poorly understood. In the present study, quantitative proteomics was employed to identify key genes associated with PEDV infection. We established an in vivo PEDV infection model using piglets and conducted in vitro studies employing Z-nucleic acid (NA)-binding protein 1 (ZBP1) knockdown and knockout (KO) models in Vero E6 cells. Several techniques were used, including transmission electron microscopy, H&E staining, confocal laser scanning microscopy, TUNEL staining, and AO/EB staining, to assess morphological changes in the intestinal tissue of piglets and to evaluate alterations in oxidative stress, mitochondrial membrane potential, and PANoptosis-related marker molecules in cells. Our findings indicated that PEDV infection results in increased expression of ZBP1 and PANoptosis-related markers. In vitro experiments demonstrated that PEDV-N colocalizes with Z-RNA and ZBP1, and that oxidative stress inhibitors effectively mitigate PEDV-induced PANoptosis. Collectively, our results suggest that ZBP1 triggers cellular oxidative damage by recognizing Z-NA structures during PEDV invasion, thereby inducing apoptosis, pyroptosis, and necroptosis, which ultimately leads to intestinal PANoptosis. These findings provide a theoretical framework for understanding PEDV-induced intestinal injury in piglets and offer valuable insights for comparative medicine research.