Endothelial cell iron overload and ferroptosis mediate thrombosis and inflammation through the miR-32-5p/neurofibromin 2 pathway

Abstract 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...

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Main Authors: Ying Deng, Xueguang Lin, Jun Wei, Bo Chen, Huafang Yan, Bo Wang, Jialong Li, Yuqun Zhao, Bo Yu, Jingdong Tang, Shuai Jiang
Format: Article
Language:English
Published: BMC 2025-06-01
Series:European Journal of Medical Research
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Online Access:https://doi.org/10.1186/s40001-025-02716-y
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author Ying Deng
Xueguang Lin
Jun Wei
Bo Chen
Huafang Yan
Bo Wang
Jialong Li
Yuqun Zhao
Bo Yu
Jingdong Tang
Shuai Jiang
author_facet Ying Deng
Xueguang Lin
Jun Wei
Bo Chen
Huafang Yan
Bo Wang
Jialong Li
Yuqun Zhao
Bo Yu
Jingdong Tang
Shuai Jiang
author_sort Ying Deng
collection DOAJ
description Abstract 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.
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spelling doaj-art-b76ce5ce54204946a2860f2ca99d78742025-08-20T02:30:45ZengBMCEuropean Journal of Medical Research2047-783X2025-06-0130111310.1186/s40001-025-02716-yEndothelial cell iron overload and ferroptosis mediate thrombosis and inflammation through the miR-32-5p/neurofibromin 2 pathwayYing Deng0Xueguang Lin1Jun Wei2Bo Chen3Huafang Yan4Bo Wang5Jialong Li6Yuqun Zhao7Bo Yu8Jingdong Tang9Shuai Jiang10Department of General Surgery, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Shanghai Key Laboratory of Vascular Lesions Regulation and RemodelingDepartment of General Surgery, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Shanghai Key Laboratory of Vascular Lesions Regulation and RemodelingSouthern Medical University Affiliated Fengxian HospitalDepartment of General Surgery, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Shanghai Key Laboratory of Vascular Lesions Regulation and RemodelingPhysical Examination Center, Shanghai Pudong Hospital, Fudan University Pudong Medical CenterDepartment of General Surgery, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Shanghai Key Laboratory of Vascular Lesions Regulation and RemodelingDepartment of General Surgery, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Shanghai Key Laboratory of Vascular Lesions Regulation and RemodelingShan Xi Yi Kang Vasculitis HospitalDepartment of General Surgery, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Shanghai Key Laboratory of Vascular Lesions Regulation and RemodelingDepartment of General Surgery, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Shanghai Key Laboratory of Vascular Lesions Regulation and RemodelingDepartment of General Surgery, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Shanghai Key Laboratory of Vascular Lesions Regulation and RemodelingAbstract 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.https://doi.org/10.1186/s40001-025-02716-yThromboangiitis obliteransFerroptosisIron metabolismNeurofibromin 2
spellingShingle Ying Deng
Xueguang Lin
Jun Wei
Bo Chen
Huafang Yan
Bo Wang
Jialong Li
Yuqun Zhao
Bo Yu
Jingdong Tang
Shuai Jiang
Endothelial cell iron overload and ferroptosis mediate thrombosis and inflammation through the miR-32-5p/neurofibromin 2 pathway
European Journal of Medical Research
Thromboangiitis obliterans
Ferroptosis
Iron metabolism
Neurofibromin 2
title Endothelial cell iron overload and ferroptosis mediate thrombosis and inflammation through the miR-32-5p/neurofibromin 2 pathway
title_full Endothelial cell iron overload and ferroptosis mediate thrombosis and inflammation through the miR-32-5p/neurofibromin 2 pathway
title_fullStr Endothelial cell iron overload and ferroptosis mediate thrombosis and inflammation through the miR-32-5p/neurofibromin 2 pathway
title_full_unstemmed Endothelial cell iron overload and ferroptosis mediate thrombosis and inflammation through the miR-32-5p/neurofibromin 2 pathway
title_short Endothelial cell iron overload and ferroptosis mediate thrombosis and inflammation through the miR-32-5p/neurofibromin 2 pathway
title_sort endothelial cell iron overload and ferroptosis mediate thrombosis and inflammation through the mir 32 5p neurofibromin 2 pathway
topic Thromboangiitis obliterans
Ferroptosis
Iron metabolism
Neurofibromin 2
url https://doi.org/10.1186/s40001-025-02716-y
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