Modelling ferroptosis in IPEC-J2 cells: insights into iron-dependent cell death

Ferroptosis, a form of regulated cell death characterised by iron accumulation and lipid peroxidation, plays a crucial role in various diseases. However, its mechanisms in livestock, particularly in pigs, remain unclear. In this study, we established an <i>in vitro</i> ferroptosis model using the porcine intestinal epithelial cell line IPEC-J2 to investigate ferroptosis mechanisms in the intestinal epithelium. IPEC-J2 cells were treated with hydrogen peroxide (H₂O₂), ferrous sulphate (FeSO₄), and the ferroptosis inducer erastin. Our results demonstrated that FeSO₄ successfully induced ferroptosis, as evidenced by increased lipid peroxidation markers, such as malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE). In the case of erastin, ferroptosis induction was confirmed by the increase in MDA levels, and the differential regulation of ferroptosis-related genes <i>ACSL4</i>, <i>TFR1</i>, and <i>FTH1</i> in response to FeSO₄ and erastin treatments. Co-treatment with the ferroptosis inhibitor ferrostatin-1 (fer-1) alleviated ferroptosis-induced lipid peroxidation and reduced cell death, further confirming the occurrence of ferroptosis. Conversely, H₂O₂ treatment increased oxidative stress without inducing ferroptosis-specific markers, suggesting that H₂O₂ does not trigger ferroptosis in IPEC-J2 cells. This study establishes a robust ferroptosis model in porcine intestinal epithelial cells and provides insights into the role of ferroptosis in intestinal health, offering a valuable platform for further research in the context of livestock. FeSO₄ and erastin treatments significantly increased lipid peroxidation markers and ferroptosis-related gene expressionCo-treatment with ferrostatin-1 effectively reduced ferroptosis-induced lipid peroxidation and cell death.H₂O₂ increased oxidative stress but did not induce ferroptosis-specific markers in IPEC-J2 cells. FeSO₄ and erastin treatments significantly increased lipid peroxidation markers and ferroptosis-related gene expression Co-treatment with ferrostatin-1 effectively reduced ferroptosis-induced lipid peroxidation and cell death. H₂O₂ increased oxidative stress but did not induce ferroptosis-specific markers in IPEC-J2 cells.

铁死亡（Ferroptosis）是一种以铁蓄积和脂质过氧化为特征的调节性细胞死亡形式，在多种疾病中发挥关键作用。不过，其在家畜（尤其是猪）中的作用机制仍不明确。本研究利用猪肠道上皮细胞系IPEC-J2建立了体外（<i>in vitro</i>）铁死亡模型，以探究肠道上皮中的铁死亡机制。用过氧化氢（H₂O₂）、硫酸亚铁（FeSO₄）及铁死亡诱导剂erastin处理IPEC-J2细胞。结果显示，FeSO₄可成功诱导铁死亡，其依据为丙二醛（MDA）和4-羟基壬烯醛（4-HNE）等脂质过氧化标志物水平升高。对于erastin，MDA水平升高以及FeSO₄与erastin处理后铁死亡相关基因<i>ACSL4</i>、<i>TFR1</i>和<i>FTH1</i>的差异调控证实了铁死亡的诱导。联合使用铁死亡抑制剂ferrostatin-1（fer-1）可缓解铁死亡诱导的脂质过氧化并减少细胞死亡，进一步证实了铁死亡的发生。相反，H₂O₂处理虽可增加氧化应激，但未诱导铁死亡特异性标志物，提示H₂O₂不会触发IPEC-J2细胞的铁死亡。本研究在猪肠道上皮细胞中建立了稳定的铁死亡模型，为理解铁死亡在肠道健康中的作用提供了见解，并为家畜相关领域的后续研究提供了宝贵平台。FeSO₄与erastin处理可显著升高脂质过氧化标志物水平并上调铁死亡相关基因表达；联合使用ferrostatin-1可有效减轻铁死亡诱导的脂质过氧化及细胞死亡；H₂O₂可增加IPEC-J2细胞的氧化应激，但不会诱导铁死亡特异性标志物。FeSO₄与erastin处理可显著升高脂质过氧化标志物水平并上调铁死亡相关基因表达；联合使用ferrostatin-1可有效减轻铁死亡诱导的脂质过氧化及细胞死亡；H₂O₂可增加IPEC-J2细胞的氧化应激，但不会诱导铁死亡特异性标志物。