目的 探讨黄芪甲苷联合骨髓间充质干细胞外泌体（bone marrow mesenchymal stem cells exosomes，BMSCs-Exos）对脊髓损伤细胞模型PC12细胞铁死亡的影响。方法 采用过氧化氢（hydrogen peroxide，H₂O₂）诱导PC12细胞构建脊髓损伤细胞模型，使用黄芪甲苷联合BMSCs-Exos进行干预。采用激光扫描共聚焦显微镜观察细胞对外泌体的摄取情况；流式细胞术检测细胞内活性氧（reactive oxygen species，ROS）水平；试剂盒测定丙二醛（malondialdehyde，MDA）、谷胱甘肽（glutathione，GSH）、亚铁离子（ferrous ion，Fe²⁺）和谷胱甘肽过氧化物酶4（glutathione peroxidase 4，GPX4）水平；JC-1荧光染色检测线粒体膜电位与透射电镜观察线粒体形态；运用qRT-PCR和Western blotting检测铁死亡相关因子GPX4、铁蛋白重链1（ferritin heavy chain 1，FTH1）和铁转运蛋白（ferroportin，FPN）的表达情况。结果 H₂O₂处理增加了细胞内ROS、MDA和Fe²⁺水平（P＜0.05），降低了GSH、GPX4、FTH1和线粒体膜电位（P＜0.05），诱发线粒体形态改变和铁死亡特征。BMSCs-Exos和黄芪甲苷单独及联合干预均显著降低细胞内ROS、MDA和Fe²⁺水平（P＜0.05），提升FPN、GSH、GPX4和线粒体膜电位（P＜0.05），改善线粒体形态、调节铁代谢及增强抗氧化能力。其中黄芪甲苷联合BMSCs-Exos处理效果更显著（P＜0.05）。结论 黄芪甲苷和BMSCs-Exos均能有效抑制脊髓损伤细胞模型中的细胞铁死亡，并显示出显著的神经保护作用。黄芪甲苷联合BMSCs-Exos不仅具有治疗脊髓损伤的潜力，还可能在涉及氧化应激和铁死亡的其他神经疾病和损伤中发挥治疗作用。

Objective To explores the effects of astragaloside Ⅳ combined with bone marrow mesenchymal stem cells exosomes (BMSCs-Exos) on ferroptosis in PC12 cell model of spinal cord injury. Methods Hydrogen peroxide (H₂O₂) was used to induce PC12 cells to construct an spinal cord injury cell model, with interventions by astragaloside Ⅳ combined with BMSCs-Exos. Laser scanning confocal microscopy was employed to observe the uptake of exosomes by the cells. Flow cytometry was used to detect intracellular reactive oxygen species (ROS) levels. Kits were utilized to measure levels of malondialdehyde (MDA), glutathione (GSH), ferrous ion (Fe²⁺), and glutathione peroxidase 4 (GPX4). JC-1 fluorescence staining was applied to detect mitochondrial membrane potential, and transmission electron microscopy was used to observe mitochondrial morphology. The expression levels of ferroptosis-related factors, including GPX4, ferritin heavy chain 1 (FTH1), and ferroportin (FPN), were analyzed using qRT-PCR and Western blotting.Results H₂O₂ treatment increased the levels of ROS, MDA, and Fe²⁺(P < 0.05), while decreasing GSH, GPX4, FTH1, and mitochondrial membrane potential (P < 0.05), leading to mitochondrial morphological changes and characteristics of ferroptosis. Both BMSCs-Exos and astragaloside Ⅳ, individually and in combination, significantly reduced ROS, MDA, and Fe²⁺ levels (P < 0.05), increased FPN, GSH, GPX4, and mitochondrial membrane potential (P < 0.05), improved mitochondrial morphology, regulated iron metabolism, and enhanced antioxidant capacity. The combined treatment of astragaloside Ⅳ and BMSCs-Exos showed a more significant effect (P < 0.05). Conclusion Astragaloside Ⅳ and BMSCs-Exos can effectively inhibit ferroptosis in the spinal cord injury cell model and demonstrate significant neuroprotective effects. The combination of astragaloside Ⅳ and BMSCs-Exos not only holds potential for the treatment of spinal cord injury but may also exhibit similar therapeutic effects in other neurological diseases and injuries involving oxidative stress and ferroptosis.

R285.5

湖南省自然科学基金面上项目（2023JJ30472）;湖南省教育厅重点课题（22A0261）;湖南省卫生健康委科研项目（B202304078103）;湖南省中医药管理局课题（B2023028）