来自fndc5工程骨髓间质干细胞的外泌体鸢尾素通过抑制YAP/EGR1/ acsl4介导的铁凋亡改善缺血性卒中。

IF 4.6 2区医学 Q1 NEUROSCIENCES

Experimental Neurology Pub Date : 2025-01-29 DOI:10.1016/j.expneurol.2025.115172

Cuini Fang, Lei Huang, Jiayi Gu, Tao Song

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引用次数: 0

摘要

背景：骨髓间充质干细胞来源的外泌体在缺血性卒中中起重要作用。鸢尾素及其前体纤维连接蛋白III型结构域含蛋白5 （FNDC5）参与神经保护作用。我们旨在阐明fndc5过表达的骨髓间充质干细胞外泌体鸢尾素在缺血性卒中中的作用。方法：建立氧糖剥夺再氧合（OGD/R）神经元细胞（HT-22和neuro2 A细胞）模型和大脑中动脉闭塞（MCAO）小鼠模型。外泌体从fndc5过表达的骨髓间充质干细胞（BMSCs- fndc5 -exos）中分离出来。用CCK-8测定细胞活力。采用商品化试剂盒检测Fe2+、乳酸脱氢酶（LDH）、谷胱甘肽（GSH）和丙二醛（MDA）。流式细胞术检测脂质活性氧（ROS）含量。采用改良神经严重程度评分（mNSS）、TTC染色和透射电镜（TEM）评估神经功能障碍、梗死体积和线粒体损伤。采用染色质免疫沉淀和双荧光素酶测定验证分子相互作用。Western blot、RT-qPCR、ELISA检测相关基因和蛋白。结果：OGD/ r处理细胞中YAP、EGR1、ACSL4表达升高。来自BMSCs-FNDC5-exos的鸢尾素可提高细胞活力并抑制铁下垂。EGR1通过转录上调ACSL4并促进OGD/ r诱导的铁下垂。此外，YAP通过转录上调EGR1，促进OGD/ r诱导的铁下垂。EGR1或YAP过表达可逆转BMSCs-FNDC5-exos的作用。EGR1沉默或BMSCs-FNDC5-exos可逆转YAP过表达诱导的铁下沉，同时EGR1沉默可进一步增强BMSCs-FNDC5-exos的诱导作用。BMSCs-FNDC5-exos减少MCAO小鼠脑梗死，改善神经功能障碍，抑制铁吊，下调YAP， EGR1, ACSL4，上调鸢尾素。结论：fndc5过表达的骨髓间质干细胞外泌体鸢尾素通过抑制YAP/EGR1/ acsl4介导的铁凋亡而改善缺血性卒中，为探索缺血性卒中治疗新策略提供了新途径。

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Exosomal irisin from FNDC5-engineered BMSCs improves ischemic stroke via inhibiting YAP/EGR1/ACSL4-mediated ferroptosis

Background

BMSCs-derived exosomes play an important role in ischemic stroke. Irisin and its precursor fibronectin type III domain-containing protein 5 (FNDC5) are implicated in neuroprotective effect. We aimed to clarify the role of exosomal irisin from FNDC5-overexpressed BMSCs in ischemic stroke.

Methods

Oxygen-glucose deprivation and reoxygenation (OGD/R) neuronal cells (HT-22 and Neuro-2a cells) model and middle cerebral artery occlusion (MCAO) mice model were established. Exosomes were isolated from FNDC5-overexpressed BMSCs (BMSCs-FNDC5-exos). Cell viability was estimated with CCK-8. Fe²⁺, lactate dehydrogenase (LDH), glutathione (GSH) and malondialdehyde (MDA) were determined with commercial kits. Moreover, lipid reactive oxygen species (ROS) was analyzed using flow cytometry. Neurological dysfunction, infarct volume and mitochondria injury were estimated with modified neurological severity score (mNSS), TTC staining and transmission electron microscopy (TEM). Chromatin immunoprecipitation and dual luciferase assay were applied to verify the molecular interactions. Western blot, RT-qPCR and ELISA were performed for the detection of related genes and proteins.

Results

YAP, EGR1 and ACSL4 were increased in OGD/R-subjected cells. Irisin from BMSCs-FNDC5-exos elevated cell viability and suppressed ferroptosis. EGR1 transcriptionally upregulated ACSL4 and promoted OGD/R-induced ferroptosis. Additionally, YAP transcriptionally upregulated EGR1 and promoted OGD/R-induced ferroptosis. EGR1 or YAP overexpression could reverse the effects of BMSCs-FNDC5-exos. EGR1 silencing or BMSCs-FNDC5-exos overturned the facilitated ferroptosis induced by YAP overexpression, meanwhile, EGR1 silencing further enhanced the effect elicited by BMSCs-FNDC5-exos. BMSCs-FNDC5-exos reduced cerebral infarction, improved neurological impairment, inhibited ferroptosis, downregulated YAP, EGR1, ACSL4 and up-regulated irisin in MCAO mice.

Conclusion

Exosomal irisin from FNDC5-overexpressed BMSCs improves ischemic stroke via inhibiting YAP/EGR1/ACSL4-mediated ferroptosis, which shed light on discovering new strategy against ischemic stroke.

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来源期刊

Experimental Neurology 医学-神经科学

CiteScore

10.10

自引率

3.80%

发文量

258

审稿时长

42 days

期刊介绍： Experimental Neurology, a Journal of Neuroscience Research, publishes original research in neuroscience with a particular emphasis on novel findings in neural development, regeneration, plasticity and transplantation. The journal has focused on research concerning basic mechanisms underlying neurological disorders.