Lim Seung Joon, Shin Sangsu, Kim Tae Hyun, Lee Sang In
Department of Animal Science and Biotechnology, Kyungpook National University, Sangju 37224, Republic of Korea.
Research Institute for Innovative Animal Science, Kyungpook National University, Sangju 37224, Republic of Korea.
Int J Mol Sci. 2025 Aug 12;26(16):7773. doi: 10.3390/ijms26167773.
Citrinin (CTN), a mycotoxin commonly found in contaminated food and animal feed, impairs intestinal barrier integrity through oxidative stress and cytotoxicity. However, its link to ferroptosis, an iron-dependent form of regulated cell death, remains unclear. This study investigated whether CTN induces ferroptosis in intestinal epithelial cells and evaluated the protective role of (EH) against CTN-induced oxidative damage and tight junction (TJ) disruption. Using IPEC-J2 cells exposed to CTN, intracellular ferrous ion (Fe) levels, reactive oxygen species (ROS) accumulation, and TJ integrity were assessed using FerroOrange and DCFH-DA staining, RT-qPCR, immunofluorescence, and WST-1 assays. Additionally, a high-throughput screen of 459 natural products identified EH extract as a top candidate in mitigating CTN toxicity. The CTN treatment significantly elevated intracellular Fe and ROS levels, downregulated antioxidant genes (notably ), and disrupted expression and TJ morphology in IPEC-J2 cells, all hallmarks of ferroptosis-like cell death. Co-treatment with EH extract effectively reversed these effects, restoring antioxidant gene expression, reducing Fe and ROS accumulation, and preserving TJ structure. Phytochemical profiling of EH extract revealed several bioactive compounds potentially responsible for its protective effects. These findings suggest that CTN induces ferroptosis-related cytotoxicity in IPEC-J2 cells, but EH alleviates this toxicity by modulating oxidative stress and iron homeostasis, supporting its potential use as a natural feed additive for intestinal protection.
桔霉素（CTN）是一种常见于受污染食品和动物饲料中的霉菌毒素，它通过氧化应激和细胞毒性损害肠道屏障完整性。然而，其与铁死亡（一种铁依赖性的程序性细胞死亡形式）之间的联系仍不清楚。本研究调查了CTN是否在肠上皮细胞中诱导铁死亡，并评估了（EH）对CTN诱导的氧化损伤和紧密连接（TJ）破坏的保护作用。使用暴露于CTN的IPEC-J2细胞，通过FerroOrange和DCFH-DA染色、RT-qPCR、免疫荧光和WST-1测定法评估细胞内亚铁离子（Fe）水平、活性氧（ROS）积累和TJ完整性。此外，对459种天然产物的高通量筛选确定EH提取物是减轻CTN毒性的最佳候选物。CTN处理显著提高了IPEC-J2细胞内的Fe和ROS水平，下调了抗氧化基因（特别是），并破坏了表达和TJ形态，这些都是铁死亡样细胞死亡的标志。与EH提取物共同处理有效地逆转了这些影响，恢复了抗氧化基因表达，减少了Fe和ROS积累，并保留了TJ结构。EH提取物的植物化学分析揭示了几种可能对其保护作用负责的生物活性化合物。这些发现表明，CTN在IPEC-J2细胞中诱导铁死亡相关的细胞毒性，但EH通过调节氧化应激和铁稳态减轻这种毒性，支持其作为肠道保护的天然饲料添加剂的潜在用途。
