Sun Xin-Yue, Wang Jing, Tian Wen-Jun, Zhang Yan-Ru, Zhang Yin-Long, Shi Yun-Long, Gong Ran-Ran, Wu Si-Nong, Qu Hong-Yu, Wang Xiao-Jia
State Key Laboratory of Veterinary Public Health and Safety, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, China.
State Key Laboratory of Veterinary Public Health and Safety, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, China; Xianyang Polytechnic Institute, Shaanxi, 712000, China.
Free Radic Biol Med. 2025 Nov;239:386-405. doi: 10.1016/j.freeradbiomed.2025.07.037. Epub 2025 Aug 5.
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.
猪流行性腹泻病毒（PEDV）是一种感染猪的冠状病毒，可导致仔猪高死亡率，对养猪业构成重大威胁。然而，PEDV诱导肠道损伤的生物学机制以及氧化应激在这一过程中的作用仍知之甚少。在本研究中，采用定量蛋白质组学来鉴定与PEDV感染相关的关键基因。我们使用仔猪建立了体内PEDV感染模型，并在Vero E6细胞中利用Z核酸（NA）结合蛋白1（ZBP1）敲低和敲除（KO）模型进行了体外研究。采用了几种技术，包括透射电子显微镜、苏木精和伊红（H&E）染色、共聚焦激光扫描显微镜、末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）染色和吖啶橙/溴化乙锭（AO/EB）染色，以评估仔猪肠道组织的形态变化，并评估细胞中氧化应激、线粒体膜电位和PANoptosis相关标记分子的改变。我们的研究结果表明，PEDV感染导致ZBP1和PANoptosis相关标记物的表达增加。体外实验表明，PEDV-N与Z-RNA和ZBP1共定位，氧化应激抑制剂可有效减轻PEDV诱导的PANoptosis。总的来说，我们的结果表明，ZBP1在PEDV入侵过程中通过识别Z-NA结构触发细胞氧化损伤，从而诱导凋亡、焦亡和坏死性凋亡，最终导致肠道PANoptosis。这些发现为理解PEDV诱导的仔猪肠道损伤提供了理论框架，并为比较医学研究提供了有价值的见解。
