中国神经再生研究(英文版) ›› 2023, Vol. 18 ›› Issue (10): 2229-2236.doi: 10.4103/1673-5374.369117

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

再灌注对大鼠缺氧缺血性脑损伤的影响

  

  • 出版日期:2023-10-15 发布日期:2023-03-28
  • 基金资助:
    国家自然科学基金项目(82271747);浙江省医疗卫生科学技术计划项目(2023RC048)

Reperfusion after hypoxia-ischemia exacerbates brain injury with compensatory activation of the anti- ferroptosis system: based on a novel rat model 

Tian-Lei Zhang1, #, Zhi-Wei Zhang2, #, Wei Lin1, Xin-Ru Lin2, Ke-Xin Lin2, Ming-Chu Fang1, Jiang-Hu Zhu1, 3, *, Xiao-Ling Guo1, 4, 5, *, Zhen-Lang Lin1, 2, 3, 4, *#br#   

  1. 1Department of Pediatrics, the Second School of Medicine, the Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, Wenzhou, Zhejiang Province, China; 2Second Clinical Medical College, Wenzhou Medical University, Wenzhou, Zhejiang Province, China; 3Key Laboratory of Children Genitourinary Diseases of Wenzhou, the Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, Wenzhou, Zhejiang Province, China; 4Key Laboratory of Perinatal Medicine of Wenzhou, the Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, Wenzhou, Zhejiang Province, China; 5Basic Medical Research Center, the Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, Wenzhou, Zhejiang Province, China
  • Online:2023-10-15 Published:2023-03-28
  • Contact: Zhen-Lang Lin, lzlprof2021@163.com; Xiao-Ling Guo, guoxling@hotmail.com; Jiang-Hu Zhu, zhujianghu@wmu.edu.cn.
  • Supported by:
    This study was supported by the National Natural Science Foundation of China, No. 82271747 (to ZLL); and Medical and Health Science and Technology Program of Zhejiang Province of China, No. 2023RC048 (to WL).

摘要:

为更好地了解缺氧缺血性脑病的病理生理机制,实验拟比较新生儿缺氧缺血再灌注脑损伤与单纯缺氧缺血脑损伤的差异。首先在传统缺氧缺血性脑病(Rice-Vannucci)模型的基础上,利用颈总动脉肌桥开发了缺氧缺血性再灌注脑损伤模型。然后以串联质量标记蛋白组学分析检验缺氧缺血性再灌注脑损伤模型和传统缺氧缺血性脑病模型中差异表达的蛋白,得出大多数差异蛋白都集中在线粒体中。进而以透射电镜观察2种模型海马CA1区线粒体,可见均出现萎缩、膜破裂以及线粒体嵴减少或消失等铁死亡的特征。紧接着以胶质纤维酸性蛋白和髓磷脂碱性蛋白作为缺氧缺血性脑病新生儿缺氧缺血性脑损伤的生物指标,结果证实了缺氧缺血性再灌注脑损伤新生大鼠模型的脑损伤并不比单纯缺氧缺血脑损伤轻。最后发现与单纯缺氧缺血性脑损伤相比,缺氧缺血性再灌注脑损伤后脑组织中铁死亡相关蛋白铁蛋白重链和谷胱甘肽过氧化酶4升高。上述结果提示缺氧缺血性再灌注脑损伤可以更好地模拟再灌注阶段的病理生理过程,且其再灌注过程在加重缺氧缺血性脑损伤的同时激活了细胞抗铁死亡系统。

https://orcid.org/0000-0002-8579-8482 (Zhen-Lang Lin); https://orcid.org/0000-0003-3153-6249 (Xiao-Ling Guo); 

https://orcid.org/0000-0002-3661-7999 (Jiang-Hu Zhu); https://orcid.org/0000-0001-8746-2985 (Tian-Lei Zhang);

 https://orcid.org/0000-0002-9768-0836 (Zhi-Wei Zhang); https://orcid.org/0000-0003-2163-8006 (Wei Lin); 

https://orcid.org/0000-0001-5488-0881 (Xin-Ru Lin); https://orcid.org/0000-0003-4336-1191 (Ke-Xin Lin); 

https://orcid.org/0000-0002-1843-3547 (Ming-Chu Fang)

关键词: 缺氧缺血性脑病, Rice-Vannucci模型, 缺氧缺血再灌注脑损伤, 模型, 蛋白质组学, 线粒体, 铁死亡, 缺氧缺血性脑损伤, 再灌注, 透射电镜

Abstract: Hypoxic-ischemic encephalopathy, which predisposes to neonatal death and neurological sequelae, has a high morbidity, but there is still a lack of effective prevention and treatment in clinical practice. To better understand the pathophysiological mechanism underlying hypoxic-ischemic encephalopathy, in this study we compared hypoxic-ischemic reperfusion brain injury and simple hypoxic-ischemic brain injury in neonatal rats. First, based on the conventional Rice-Vannucci model of hypoxic-ischemic encephalopathy, we established a rat model of hypoxic-ischemic reperfusion brain injury by creating a common carotid artery muscle bridge. Then we performed tandem mass tag-based proteomic analysis to identify differentially expressed proteins between the hypoxic-ischemic reperfusion brain injury model and the conventional Rice-Vannucci model and found that the majority were mitochondrial proteins. We also performed transmission electron microscopy and found typical characteristics of ferroptosis, including mitochondrial shrinkage, ruptured mitochondrial membranes, and reduced or absent mitochondrial cristae. Further, both rat models showed high levels of glial fibrillary acidic protein and low levels of myelin basic protein, which are biological indicators of hypoxic-ischemic brain injury and indicate similar degrees of damage. Finally, we found that ferroptosis-related Ferritin (Fth1) and glutathione peroxidase 4 were expressed at higher levels in the brain tissue of rats with hypoxic-ischemic reperfusion brain injury than in rats with simple hypoxic-ischemic brain injury. Based on these results, it appears that the rat model of hypoxic-ischemic reperfusion brain injury is more closely related to the pathophysiology of clinical reperfusion. Reperfusion not only aggravates hypoxic-ischemic brain injury but also activates the anti-ferroptosis system. 

Key words: ferroptosis, hypoxic-ischemic brain injury, hypoxic-ischemic encephalopathy, hypoxic-ischemic reperfusion brain injury, mitochondria, model, proteomic analysis, reperfusion, Rice-Vannucci, transmission electron microscopy