南方医科大学学报

南方医科大学学报 ›› 2025, Vol. 45 ›› Issue (10): 2071-2081.doi: 10.12122/j.issn.1673-4254.2025.10.03

• • 上一篇    

天麻素通过激活GPX4/SLC7A11/FTH1信号抑制铁死亡减轻新生小鼠缺氧缺血性脑损伤

郭涛1,2(), 陈柏霖1, 石金沙1, 匡显锋1, 余腾跃1, 魏嵩1, 刘雄2, 肖蓉1, 李娟娟1()   

  1. 1.昆明医科大学基础医学院人体解剖学与组织胚胎学系，云南 昆明 650500
    2.昆明理工大学基础医学院解剖学教研室，云南 昆明 650500
  • 收稿日期:2025-06-19 出版日期:2025-10-20 发布日期:2025-10-24
  • 通讯作者: 李娟娟 E-mail:475612496@qq.com;lijuanjuan@kmmu.edu.cn
  • 作者简介:郭 涛，实验师，在读博士研究生，E-mail: 475612496@qq.com
  • 基金资助:
    国家自然科学基金(32460218);国家自然科学基金(31960194);昆明医科大学博士研究生创新基金(J114150330296);昆明医科大学大学生创新训练计划基金(2024CYD038);昆明医科大学大学生创新训练计划基金(2024CYD161)

Gastrodin inhibits ferroptosis to alleviate hypoxic-ischemic brain damage in neonatal mice by activating GPX4/SLC7A11/FTH1 signaling

Tao GUO1,2(), Bolin CHEN1, Jinsha SHI1, Xianfeng KUANG1, Tengyue YU1, Song WEI1, Xiong LIU2, Rong XIAO1, Juanjuan LI1()   

  1. 1.Department of Human Anatomy & Histology and Embryology, School of Basic Medical Sciences, Kunming Medical University, Kunming 650500, China
    2.Department of Anatomy, School of Basic Medical Sciences, Kunming University of Science and Technology, Kunming 650500, China
  • Received:2025-06-19 Online:2025-10-20 Published:2025-10-24
  • Contact: Juanjuan LI E-mail:475612496@qq.com;lijuanjuan@kmmu.edu.cn
  • Supported by:
    National Natural Science Foundation of China(32460218)

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摘要：

目的 探讨天麻素（GAS）通过调控GPX4/SLC7A11/FTH1信号通路抑制神经元铁死亡的机制，并评估其在新生小鼠缺氧缺血性脑损伤（HIBD）中的治疗效果。 方法 选取24只出生后9~11 d的C57BL/6J新生小鼠，采用右侧颈总动脉结扎缺血，并置于92% N2和8% O2的环境中缺氧1 h，以建立HIBD模型。将小鼠随机分为假手术组（Sham）、缺氧缺血模型组（HIBD）、天麻素低剂量组（HIBD+GAS-L，100 mg/kg）和天麻素高剂量组（HIBD+GAS-H，200 mg/kg），6只/组。持续腹腔给药3 d后，选取小鼠脑皮层缺血半暗带组织，通过HE染色、Nissl 染色、Zea-Longa法评分检测其病理损伤，评估亚铁离子、丙二醛（MDA）、谷胱甘肽（GSH）、二氢乙啶（DHE）的水平和铁死亡相关蛋白GPX4、SLC7A11和FTH1的表达，并电镜检测铁死亡的形态学特征变化。体外实验选用HT22神经细胞，添加天麻素及GPX4的抑制剂RSL3干预1 h，并进行缺氧缺糖2 h，分为对照组（Control）、氧糖剥夺组（OGD）、天麻素治疗组（OGD+GAS，0.5 mmol/L）、RSL3干预组（OGD+RSL3，10 μmol/L）、天麻素和RSL3联合组（OGD+GAS+RSL3）。通过Western blotting和免疫荧光双标染色检测铁死亡相关蛋白的表达变化，同时评价FerroOrange、DCFH-DA、BODIPY-C11、MDA、GSH、JC-1和细胞活性指标。 结果 天麻素低剂量和高剂量组均表现出改善HIBD病理损伤及神经行为学评分的效果（P<0.05），铁离子、脂质过氧化物及ROS的水平降低，抗氧化能力得到恢复（P<0.05），电镜观察到线粒体的脊得到恢复。Western blotting和免疫荧光显示，天麻素的这种神经保护作用是通过激活GPX4、SLC7A11和FTH1信号的表达水平而实现的。此外，细胞模型得到了类似的结果，并且当GPX4信号被RSL3靶向抑制后，天麻素对Fe2+、ROS、BODIPY-C11和MDA的抑制作用被RSL3抵消（P<0.05），同时，天麻素对GSH、细胞活力和线粒体膜电位水平的增强效果被RSL3减弱（P<0.05）。 结论 天麻素通过上调GPX4/SLC7A11/FTH1信号通路，抑制神经元的铁死亡，从而在新生小鼠HIBD中发挥神经保护作用。

关键词: 天麻素, 缺氧缺血性脑损伤, 氧糖剥夺, 铁死亡, GPX4/SLC7A11/FTH1, 神经元

Abstract:

Objective To evaluate the therapeutic effect of gastrodin against hypoxic-ischemic brain damage (HIBD) in neonatal mice and explore the role of GPX4/SLC7A11/FTH1 signaling in mediating its effect. Methods Twenty-four 9- to 11-day-old C57BL/6J mice were randomized equally into 4 groups for sham operation, HIBD modeling by right common carotid artery ligation and subsequent exposure to hypoxia for 1 h, or gastrodin treatment at 100 or 200 mg/kg before and at 1 and 2 days after modeling. The mice then underwent neurological assessment (Zea-Longa scores), and the cerebral cortical penumbra tissue were collected for HE and Nissl staining, detection of ferroptosis biomarkers and protein expressions of GPX4, SLC7A11, and FTH1 with Western blotting and immunofluorescence co-localization, and observation of mitochondrial ultrastructure with electron microscopy. In cultured HT22 neuronal cells with oxygen-glucose deprivation (OGD) for 2 h, the effects of pretreatments with 0.5 mmol/L gastrodin, 10 μmol/L RSL3 (a GPX4 inhibitor), alone or in combination, were analyzed on expressions of ferroptosis-related proteins, cellular Fe²⁺, ROS, lipid peroxidation, MDA, and GSH levels, mitochondrial membrane potential (JC-1), and cell viability. Results Gastrodin treatment at the two doses both significantly ameliorated HIBD and neurological deficits of the mice, reduced mitochondrial damage and Fe²⁺, MDA and ROS levels, increased GSH level, and upregulated GPX4, SLC7A11, and FTH1 protein expressions. In HT22 cells, gastrodin pretreatment obviously attenuated OGD-induced ferroptosis and improved cell viability and mitochondrial function. Co-treatment with RSL3 potently abrogated the inhibitory effects of gastrodin on Fe²⁺, ROS, BODIPY-C11, and MDA levels and attenuated its protective effects on GSH level, cell viability, and mitochondrial membrane potential. Conclusion Gastrodin provides neuroprotective effects in neonatal mice with HIBD by suppressing neuronal ferroptosis via upregulating the GPX4/SLC7A11/FTH1 signaling pathway.

Key words: gastrodin, hypoxic-ischemic brain damage, oxygen-glucose deprivation, ferroptosis, GPX4/SLC7A11/FTH1, neurons