Deng Ying, Lin Xueguang, Wei Jun, Chen Bo, Yan Huafang, Wang Bo, Li Jialong, Zhao Yuqun, Yu Bo, Tang Jingdong, Jiang Shuai
Department of General Surgery, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Shanghai Key Laboratory of Vascular Lesions Regulation and Remodeling, Shanghai, 201399, China.
Fudan Zhangjiang Institute, Shanghai, China.
Eur J Med Res. 2025 Jun 6;30(1):463. doi: 10.1186/s40001-025-02716-y.
Thromboangiitis obliterans (TAO) is characterized by progressive inflammatory vasculopathy featuring thrombotic occlusion. Aberrant thrombosis induces endothelial damage through pathological clotting, while iron may act as a pro-oxidant cofactor. However, the function and mechanism of iron in TAO pathogenesis and endothelial damage remain to be elucidated. In the current study, the iron status and key lipid peroxidation markers (MDA, 4HNE, and ACSL4) were evaluated in patients with TAO and the sodium laurate-induced rat model. The CCK-8 assay, immunofluorescence, western blot, qPCR, and transmission electron microscopy were employed to detect iron overload and ferroptosis in vascular endothelial cells. In addition, bioinformatics analysis, luciferase reporter gene assay, qPCR, and western blot were used to confirm the miR-32-5p/Neurofibromin-2 (NF2) pathway in vitro. The therapeutic feasibility was validated by deferoxamine and Ferrostatin-1 treatment in vivo. The results showed iron overload and increased TFR1 expression in the vessel lesions of patients with TAO, as well as significant increases in MDA, 4HNE, and ACSL4. Serum from patients with TAO increased intracellular iron and lipid peroxidation and decreased the viability of HUVECs in vitro. Mechanism studies indicated that exosomal miR-32-5p increased in patients with TAO and could target and decrease the expression of NF2, which then decreased the phosphorylation of YAP at Ser109 and Ser217 sites. Then the NF2-targeted genes TFR1 and ACSL4 were upregulated. Finally, deferoxamine and Ferrostatin-1 treatment relieved the disease score, inflammation, and ferroptosis in vivo. This study newly demonstrates that iron overload and ferroptosis are key risk factors in patients with TAO and that the exosomal miR-32-5p/NF2 pathway may play an important role in TAO pathogenesis.
血栓闭塞性脉管炎（TAO）的特征是一种以血栓形成性闭塞为特征的进行性炎症性血管病变。异常血栓形成通过病理性凝血诱导内皮损伤，而铁可能作为一种促氧化辅因子。然而，铁在TAO发病机制和内皮损伤中的作用及机制仍有待阐明。在本研究中，对TAO患者和月桂酸钠诱导的大鼠模型的铁状态及关键脂质过氧化标志物（丙二醛、4-羟基壬烯醛和长链脂酰辅酶A合成酶4）进行了评估。采用CCK-8法、免疫荧光法、蛋白质印迹法、定量聚合酶链反应（qPCR）和透射电子显微镜检测血管内皮细胞中的铁过载和铁死亡。此外，利用生物信息学分析、荧光素酶报告基因检测、qPCR和蛋白质印迹在体外证实了miR-32-5p/神经纤维瘤蛋白2（NF2）通路。通过去铁胺和铁死亡抑制剂1在体内的治疗验证了其可行性。结果显示，TAO患者血管病变中铁过载且转铁蛋白受体1（TFR1）表达增加，同时丙二醛、4-羟基壬烯醛和长链脂酰辅酶A合成酶4显著增加。TAO患者的血清在体外增加了细胞内铁和脂质过氧化，并降低了人脐静脉内皮细胞（HUVEC）的活力。机制研究表明，TAO患者中外泌体miR-32-5p增加，可靶向并降低NF2的表达，进而降低Yes相关蛋白（YAP）在丝氨酸109和丝氨酸217位点的磷酸化。然后，NF2靶向基因TFR1和长链脂酰辅酶A合成酶4上调。最后，去铁胺和铁死亡抑制剂1治疗减轻了体内疾病评分、炎症和铁死亡。本研究首次证明铁过载和铁死亡是TAO患者的关键危险因素，且外泌体miR-32-5p/NF2通路可能在TAO发病机制中起重要作用。
