FTO-mediated m6A Demethylation of OTUB1 stabilizes SLC7A11 to alleviate Ferroptosis in cerebral ischemia/reperfusion injury

IF 2 4区医学 Q3 NEUROSCIENCES

Journal of Stroke & Cerebrovascular Diseases Pub Date : 2025-04-13 DOI:10.1016/j.jstrokecerebrovasdis.2025.108316

Youjin Shen , Wentao Liu , Zonghua Zhou , Jianwen He , Xiaokun Qi

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引用次数: 0

Abstract

Backgroud

Therapeutic strategies for cerebral ischemia/reperfusion (I/R) injury, an important contributor to neurological impairment and disability, exhibit limited efficacy. Reperfusion therapy intensifies neuronal damage by promoting iron deposition, ferroptosis (lipid peroxidation-associated iron-dependent cellular death), and reactive oxygen species (ROS) accumulation.

Methods

we investigated the role of the m6A demethylase FTO in modulating ferroptosis during cerebral I/R injury, using middle cerebral artery occlusion/reperfusion (MCAO/R) model rats and neuronal cells subjected to oxygen glucose deprivation/reoxygenation (OGD/R) as in vivo and in vitro experimental platforms, respectively. Neurological scores and cerebral infarction volumes were measured by TTC staining. FTO, OTUB1, and SLC7A11 levels, and FTO demethylase activity, were assessed by qRT-PCR, western blotting, and immunohistochemistry. MeRIP was applied to ascertain the m6A methylation status of OTUB1 mRNA. Apoptotic rates and cell viability were quantitatively aalyzed by flow cytometry and CCK-8 assay, respectively, while brain tissue apoptosis was evaluated using TUNEL staining.

Results

MCAO/R rat brains and OGD/R cells showed decreased FTO expression and increased OTUB1 m6A methylation. FTO overexpression upregulated OTUB1 by diminishing m6A methylation, consequently stabilizing SLC7A11 and reducing ferroptosis. FTO or OTUB1 silencing increased ferroptosis, while their co-overexpression enhanced neuroprotective effects. FTO overexpression reduced infarct volume and cell apoptosis, and improved neurological outcomes in vivo.

Conclusions

FTO enhanced OTUB1 expression via m6A demethylation, stabilizing SLC7A11, and inhibiting ferroptosis to alleviate cerebral I/R injury. The FTO/OTUB1/SLC7A11 pathway is a viable therapeutic target for ischemic stroke, providing novel perspectives on the molecular mechanisms underlying neuroprotection and proposing innovative m6A-based therapies.

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FTO 介导的 OTUB1 m6A 去甲基化可稳定 SLC7A11，从而缓解脑缺血再灌注损伤中的铁变态反应

脑缺血/再灌注（I/R）损伤是神经功能损伤和残疾的重要因素，其治疗策略的疗效有限。再灌注治疗通过促进铁沉积、铁凋亡（脂质过氧化相关的铁依赖性细胞死亡）和活性氧（ROS）积累而加剧神经元损伤。方法采用大脑中动脉闭塞/再灌注（MCAO/R）模型大鼠和氧葡萄糖剥夺/再氧化（OGD/R）神经细胞作为体内和体外实验平台，研究m6A去甲基化酶FTO在脑I/R损伤中对铁上吊的调节作用。TTC染色测定神经学评分和脑梗死体积。FTO、OTUB1和SLC7A11水平以及FTO去甲基化酶活性通过qRT-PCR、western blotting和免疫组织化学进行评估。应用MeRIP确定OTUB1 mRNA的m6A甲基化状态。流式细胞术和CCK-8法分别定量分析凋亡率和细胞活力，TUNEL染色法检测脑组织凋亡。结果smcao /R大鼠脑和OGD/R细胞FTO表达降低，OTUB1 m6A甲基化升高。FTO过表达通过减少m6A甲基化而上调OTUB1，从而稳定SLC7A11并减少铁下垂。FTO或OTUB1沉默增加铁凋亡，而它们的共过表达增强了神经保护作用。FTO过表达减少了梗死体积和细胞凋亡，并改善了体内神经系统预后。结论sfto通过m6A去甲基化OTUB1表达，稳定SLC7A11，抑制铁下沉，减轻脑I/R损伤。FTO/OTUB1/SLC7A11通路是缺血性卒中的可行治疗靶点，为神经保护的分子机制提供了新的视角，并提出了基于m6的创新治疗方法。

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来源期刊

Journal of Stroke & Cerebrovascular Diseases Medicine-Surgery

CiteScore

5.00

自引率

4.00%

发文量

583

审稿时长

62 days

期刊介绍： The Journal of Stroke & Cerebrovascular Diseases publishes original papers on basic and clinical science related to the fields of stroke and cerebrovascular diseases. The Journal also features review articles, controversies, methods and technical notes, selected case reports and other original articles of special nature. Its editorial mission is to focus on prevention and repair of cerebrovascular disease. Clinical papers emphasize medical and surgical aspects of stroke, clinical trials and design, epidemiology, stroke care delivery systems and outcomes, imaging sciences and rehabilitation of stroke. The Journal will be of special interest to specialists involved in caring for patients with cerebrovascular disease, including neurologists, neurosurgeons and cardiologists.