Shi Jinsha, Xiao Xiao, Wang Zhao, Zhang Xinglin, Kuang Xianfeng, Yang Shuhan, Zuo Hanjun, Guo Tao, Xiao Rong, Li Yue, Liang Yundan, Gao Linbo, Li Juanjuan
Department of Anatomy and Histology & Embryology, School of Basic Medical Sciences, Kunming Medical University, 1168 West Chunrong Road, Kunming 650500, PR China.
Laboratory of Molecular Translational Medicine, Center for Translational Medicine, Key Laboratory of Birth Defects and Related Diseases of Women and Children (Sichuan University), Ministry of Education; NHC Key Laboratory of Chronobiology (Sichuan University); Children's Medicine Key Laboratory of Sichuan Province; West China Second University Hospital, Sichuan University, Chengdu, Sichuan 610041, PR China; Department of Medical Genetics, West China Second University Hospital, Sichuan University, Chengdu, Sichuan 610041, PR China.
Phytomedicine. 2025 Nov;147:157177. doi: 10.1016/j.phymed.2025.157177. Epub 2025 Aug 16.
Ischemic stroke (IS) remains a major contributor to global morbidity and mortality, largely due to limited therapies and unclear pathogenesis. Gastrodin (GAS), a bioactive ingredient from traditional Chinese medicine, has shown potential in mitigating apoptosis, inflammation, and pyroptosis. Recent research highlights ferroptosis as a crucial factor in IS development.
In this study, we aimed to examine the effect and mechanism of GAS on ferroptosis in IS.
Mendelian randomization (MR) was applied to evaluate the effect of GAS on IS risk. The neuroprotective and anti-ferroptotic effects of GAS were assessed through TTC staining, neurological deficit scoring, and detection of ferroptosis-related biomarkers. GAS target was predicted via network pharmacology and molecular docking, and validated using surface plasmon resonance and cellular thermal shift assay. Phosphorylation, nuclear translocation, and N⁶-methyladenosine methylation were analyzed using Western blotting, immunofluorescence, and MeRIP-qPCR.
MR analyses identified a negative association between GAS and IS. GAS treatments attenuated cerebral infarct volume, improved neurological function, and alleviated ferroptosis both in vivo and in vitro. Mechanistically, GAS binds to PI3K, activating the PI3K/Akt signaling pathway through phosphorylation. Phosphorylated Akt induces serine/threonine phosphorylation and nuclear translocation of Alkbh5, which reduces the mA methylation levels of glutamate-cysteine ligase modifier subunit (Gclm), resulting in increased Gclm expression and enhanced GSH synthesis, and ultimately contributing to the inhibition of ferroptosis in IS.
This study is the first to demonstrate that GAS mitigates IS-induced ferroptosis via the PI3K/Akt-Alkbh5-Gclm axis, bridging epigenetic regulation and iron metabolism.
缺血性中风（IS）仍然是全球发病率和死亡率的主要原因，这主要是由于治疗方法有限且发病机制不明。天麻素（GAS）是一种来自中药的生物活性成分，已显示出在减轻细胞凋亡、炎症和焦亡方面的潜力。最近的研究强调铁死亡是IS发展的关键因素。
在本研究中，我们旨在研究GAS对IS中铁死亡的影响及其机制。
采用孟德尔随机化（MR）方法评估GAS对IS风险的影响。通过TTC染色、神经功能缺损评分以及检测铁死亡相关生物标志物来评估GAS的神经保护和抗铁死亡作用。通过网络药理学和分子对接预测GAS靶点，并使用表面等离子体共振和细胞热位移分析进行验证。使用蛋白质免疫印迹法、免疫荧光法和MeRIP-qPCR分析磷酸化、核转位和N⁶-甲基腺苷甲基化。
MR分析确定GAS与IS之间存在负相关。GAS治疗在体内和体外均能减少脑梗死体积、改善神经功能并减轻铁死亡。机制上，GAS与PI3K结合，通过磷酸化激活PI3K/Akt信号通路。磷酸化的Akt诱导Alkbh5的丝氨酸/苏氨酸磷酸化和核转位，这降低了谷氨酸-半胱氨酸连接酶修饰亚基（Gclm）的mA甲基化水平，导致Gclm表达增加和谷胱甘肽合成增强，最终有助于抑制IS中的铁死亡。
本研究首次证明GAS通过PI3K/Akt-Alkbh5-Gclm轴减轻IS诱导的铁死亡，架起了表观遗传调控与铁代谢之间的桥梁。
