Abstract：
Deoxynivalenol (DON) is a secondary metabolite produced by fungi, which causes serious health issues worldwide due to its widespread presence in human and animal diets. Necroptosis is a newly proposed cell death mode and has been proposed as a potential mechanism of intestinal disease. This study aimed to investigate the role of necroptosis in intestinal damage caused by DON exposure. Piglets were fed diets with or without 4 mg/kg DON for 3 weeks or given a gavage of 2 mg/kg BW DON or sterile saline to investigate the effects of chronic or acute DON exposure on the gut, respectively. IPEC-1 cells were challenged with different concentrations of DON to investigate the effect of DON exposure on the intestinal epithelial cells (IECs) in vitro. Subsequently, the inhibitors of necroptosis were used to treat cells or piglets prior to DON challenge. Chronic and acute DON exposure both caused morphological damage, reduction of disaccharidase activity, decrease of tight junction protein expression, inflammation of the small intestine, and necroptosis of intestinal epithelial cells in piglets. Necroptosis was also detected when IPEC-1 cell damage was induced by DON in vitro. The suppression of necroptosis in IPEC-1 cells by inhibitors (necrostatin-1 (Nec-1), GSK\'872, or GW806742X) alleviated cell death, the decrease of tight junction protein expression, oxidative stress, and the inflammatory response induced by DON. Furthermore, pre-treatment with Nec-1 in piglets was also observed to protect the intestine against DON-induced enterotoxicity. Additionally, the expression of histone methyltransferase SETDB1 was abnormally downregulated upon chronic and acute DON exposure in piglets, and necroptosis was activated in IPEC-1 cells due to knockout of SETDB1. Collectively, these results demonstrate that necroptosis of IECs is a mechanism of DON-induced enterotoxicity and SETDB1 mediates necroptosis upon DON exposure in IECs, suggesting the potential for targeted inhibition of necroptosis to alleviate mycotoxin-induced enterotoxicity and intestinal disease.
摘要：
脱氧雪腐镰刀菌烯醇（DON）是真菌产生的次生代谢产物，由于其在人类和动物饮食中的广泛存在，在全球范围内引起严重的健康问题。坏死性凋亡是一种新提出的细胞死亡模式，已被认为是肠道疾病的潜在机制。本研究旨在探讨凋亡在DON暴露所致肠道损伤中的作用。仔猪饲喂有或没有4mg/kgDON的饮食3周，或给予2mg/kgBWDON或无菌盐水的灌胃，以研究慢性或急性DON暴露对肠道的影响，分别。用不同浓度的DON攻击IPEC-1细胞，以研究DON暴露对肠上皮细胞（IECs）的影响。随后，在DON攻击之前，使用坏死的抑制剂治疗细胞或仔猪。慢性和急性DON暴露均导致形态学损伤，二糖酶活性的降低，紧密连接蛋白表达减少，小肠的炎症，仔猪肠上皮细胞坏死。当体外DON诱导IPEC-1细胞损伤时，也检测到坏死。抑制剂抑制IPEC-1细胞的坏死性凋亡（necrostatin-1（Nec-1），GSK\'872，或GW806742X）减轻细胞死亡，紧密连接蛋白表达的减少，氧化应激，以及DON诱导的炎症反应。此外，在仔猪中还观察到用Nec-1预处理可以保护肠道免受DON诱导的肠毒性。此外,在仔猪慢性和急性DON暴露后，组蛋白甲基转移酶SETDB1的表达异常下调，由于SETDB1的敲除，IPEC-1细胞中的坏死被激活。总的来说，这些结果表明，IECs的坏死性凋亡是DON诱导的肠毒性的机制，SETDB1介导IECs中DON暴露后的坏死性凋亡，提示靶向抑制坏死以减轻霉菌毒素诱导的肠毒性和肠道疾病的潜力。
