中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (11): 2806-2813.doi: 10.12307/2026.038

• 组织构建基础实验 basic experiments in tissue construction • 上一篇    下一篇

内质网应激促进铁死亡加重脑缺血再灌注损伤

张月婷1，李静林2，傅振燚3，晏  斐3，高  宇3，刘佳鑫3   

  1. 1昆明医科大学第二附属医院，云南省昆明市   650100；2昆明医科大学第一附属医院，云南省昆明市   650032；3昆明理工大学医学院，云南省昆明市   650500
  • 收稿日期:2025-02-15 接受日期:2025-05-09 出版日期:2026-04-18 发布日期:2025-09-06
  • 通讯作者: 刘佳鑫，博士，讲师，昆明理工大学医学院，云南省昆明市 650500
  • 作者简介:张月婷，女，1985年生，云南省德宏州人，汉族，2018年昆明医科大学毕业，硕士，主治医师，主要从事脑缺血机制研究。
  • 基金资助:
    昆明医科大学第二附属医院内部科技项目(XJ2021003601)，项目负责人：张月婷；昆明理工大学联合专项(KUST-KH2023001Y)，项目负责人：刘佳鑫；云南省科技厅：创新引导和技术型企业培育计划(202404AM350005)，项目负责人：刘佳鑫；云南省教育厅科学研究基金项目(2023J0210)，项目负责人：李静林

Endoplasmic reticulum stress promotes ferroptosis and aggravates cerebral ischemia-reperfusion injury#br#
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Zhang Yueting1, Li Jinglin2, Fu Zhenyi3, Yan Fei3, Gao Yu3, Liu Jiaxin3   

  1. 1The Second Affiliated Hospital of Kunming Medical University, Kunming 650100, Yunnan Province, China; 2First Affiliated Hospital of Kunming Medical University, Kunming 650032, Yunnan Province, China; 3Medical School of Kunming University of Science and Technology, Kunming 650500, Yunnan Province, China
  • Received:2025-02-15 Accepted:2025-05-09 Online:2026-04-18 Published:2025-09-06
  • Contact: Liu Jiaxin, PhD, Lecturer, Medical School of Kunming University of Science and Technology, Kunming 650500, Yunnan Province, China
  • About author:Zhang Yueting, MS, Attending physician, The Second Affiliated Hospital of Kunming Medical University, Kunming 650100, Yunnan Province, China
  • Supported by:
    the Internal Science and Technology Project of the Second Affiliated Hospital of Kunming Medical University, No. XJ2021003601 (to ZYT); Kunming University of Science and Technology Joint Specialization, No. KUST-KH2023001Y (to LJX); Yunnan Provincial Department of Science and Technology: Innovation Guidance and Technology-based Enterprise Cultivation Program, No. 202404AM350005 (to LJX); Yunnan Provincial Department of Education Scientific Research Project, No. 2023J0210 (to LJL) 

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摘要：


文题释义：
铁死亡：是一种独特的细胞死亡方式，依赖于铁离子和活性氧的相互作用，导致脂质过氧化物的积累。铁死亡本质是谷胱甘肽的耗竭，谷胱甘肽过氧化物酶4活性下降，导致脂质氧化物不能被代谢，进而引发细胞死亡，主要特点包括：细胞死亡过程中伴随着大量的铁离子累积，同时还会出现脂质过氧化、活性氧升高；线粒体变小，线粒体膜皱缩，线粒体嵴减少或消失，外膜破碎，但细胞核形态变化不明显。
内质网应激：是指细胞内质网中蛋白质折叠和修饰发生异常时所引发的一种细胞应激反应。当细胞面临营养匮乏、Ca²⁺调控代谢失衡、毒素刺激、持续氧化应激等应激源时，内质网功能被部分中断，导致蛋白质加工和运输受损，未折叠或错误折叠的蛋白质在内质网内积累，从而引发内质网应激。

背景：栓子栓塞脑动脉导致相应的脑组织缺血缺氧，在恢复血流以及再氧合时，通常会出现脑缺血再灌注损伤。然而，目前关于铁死亡与内质网应激在脑缺血再灌注损伤中相互作用的研究较为有限。
目的：探讨内质网应激和铁死亡在脑缺血再灌注损伤中的作用及机制。
方法：HT-22细胞系分为4组：对照组未进行造模和干预，模型组构建氧糖剥夺/再灌注模型，4-PBA组在进行氧糖剥夺/再灌注前使用
1.25 mmol/L内质网应激抑制剂4-PBA处理2 h，4-PBA组+CCT组在进行氧糖剥夺/再灌注前使用1.25 mmol/L 4-PBA和5 μmol/L蛋白激酶RNA样ER激酶激活剂CCT020312处理2 h。通过CCK-8实验检测细胞活力；Western blot检测铁死亡、内质网应激及蛋白激酶RNA样ER激酶/转录激活因子4信号通路相关蛋白的表达；流式细胞术检测细胞凋亡；使用试剂盒检测Fe²⁺浓度。
结果与结论：与对照组相比，氧糖剥夺/再灌注处理通过诱导铁死亡、内质网应激和细胞凋亡机制显著抑制了神经细胞的增殖活性。内质网应激抑制剂4-PBA处理有效减轻了氧糖剥夺/再灌注引起的铁死亡、凋亡以及细胞活力抑制现象。此外，4-PBA还显著降低了氧糖剥夺/再灌注造模后蛋白激酶RNA样ER激酶和转录激活因子4表达水平。而PERK激活剂CCT020312则逆转了4-PBA上述作用。结果表明：内质网应激通过蛋白激酶RNA样ER激酶/转录激活因子4信号通路促进铁死亡，从而加重脑缺血再灌注损伤。
https://orcid.org/0000-0003-3508-9132 (刘佳鑫) 

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 铁死亡, 内质网应激, 急性缺血性脑卒中, 氧糖剥夺/再灌注, 细胞活力, 细胞凋亡, 蛋白激酶RNA样ER激酶(PERK), 转录激活因子4(ATF4) 

Abstract: BACKGROUND: Embolization of cerebral arteries leads to cerebral tissue ischemia and hypoxia. Restoration of blood flow and reoxygenation usually results in cerebral ischemia-reperfusion injury. However, current research on the interaction between ferroptosis and endoplasmic reticulum stress in cerebral ischemia-reperfusion injury is relatively limited.
OBJECTIVE: To investigate the roles and mechanisms of endoplasmic reticulum stress and ferroptosis in cerebral ischemia-reperfusion injury.
METHODS: The HT-22 cell line was used and divided into four groups. An oxygen glucose deprivation/reperfusion (OGD/R) model was established in the model group. Cells were treated with 1.25 mmol/L 4-PBA (an endoplasmic reticulum stress inhibitor) for 2 hours prior to OGD/R, forming the 4-PBA group. Cells were treated with 1.25 mmol/L 4-PBA and 5 μmol/L CCT020312 (a protein kinase RNA-like ER kinase activator) for 2 hours before OGD/R, forming the 4-PBA+CCT group. Cell viability was detected by cell counting kit-8 experiment; western blot was used to detect the expression of ferroptosis-, endoplasmic reticulum stress, and protein kinase RNA-like ER kinase/transcription activating factor 4-related proteins; flow cytometry was used to detect cell apoptosis; and a reagent kit was used to detect the concentration of Fe2+.
RESULTS AND CONCLUSION: Compared with the control group, OGD/R treatment significantly suppressed neuronal cell proliferation activity by inducing ferroptosis, endoplasmic reticulum stress and apoptosis mechanisms. Treatment with the endoplasmic reticulum stress inhibitor 4-PBA effectively alleviated ferroptosis, apoptosis and inhibition of cell viability caused by OGD/R. In addition, 4-PBA significantly reduced the expression levels of protein kinase RNA-like ER kinase and transcription activating factor 4 induced by OGD/R. The protein kinase RNA-like ER kinase activator CCT020312 reversed the inhibitory effects of 4-PBA on endoplasmic reticulum stress, ferroptosis, apoptosis and cell activity in the OGD/R model. To conclude, endoplasmic reticulum stress promotes ferroptosis through the protein kinase RNA-like ER kinase/transcription activating factor 4 signaling pathway, thereby exacerbating cerebral ischemia-reperfusion injury. 

Key words: ferroptosis, endoplasmic reticulum stress, acute ischemic stroke, oxygen glucose deprivation/reperfusion, cell viability, apoptosis, protein kinase RNA-like ER kinase (PERK), transcription activating factor 4 (ATF4) 

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