中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2020, Vol. 15 ›› Issue (5): 903-911.doi: 10.4103/1673-5374.268973

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

选择性脑低温对局灶性脑缺血再灌注损伤神经保护与抑制的线粒体分裂蛋白1表达有关

  

  • 出版日期:2020-05-15 发布日期:2020-06-01

Selective brain hypothermia-induced neuroprotection against focal cerebral ischemia/reperfusion injury is associated with Fis1 inhibition

Ya-Nan Tang1, Gao-Feng Zhang1, Huai-Long Chen1, Xiao-Peng Sun1, Wei-Wei Qin1, Fei Shi1, Li-Xin Sun1, Xiao-Na Xu2, Ming-Shan Wang1   

  1. 1 Department of Anesthesiology, Affiliated Qingdao Municipal Hospital of Qingdao University, Qingdao, Shandong Province, China
    2 Department of Central Laboratory, Affiliated Qingdao Municipal Hospital of Qingdao University, Qingdao, Shandong Province, China
  • Online:2020-05-15 Published:2020-06-01
  • Contact: Ming-Shan Wang, MS,wmsqingdao@163.com.
  • Supported by:
    This study was supported by the Natural Science Foundation of Shandong Province of China, No. ZR2015HM023 (to MSW); the Science and Technology Plan Project of Qingdao City of China, No. 19-6-1-50-nsh (to MSW).

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摘要：

临床研究表明，选择性脑低温是减轻脑卒中后神经元损伤的有效方法，而且可以减少全身低温所带来的并发症，然而选择性脑低温影响线粒体分裂的具体机制尚目前不清楚。为揭示选择性脑低温对局灶性脑I/R损伤时线粒体分裂系统的关键因子-线粒体分裂蛋白1表达的调节作用，实验将SD大鼠随机分为4组：sham组仅暴露颈动脉，其他3组采用线栓法建立大鼠大脑中动脉栓塞模型，大脑中动脉栓塞2 h后，I/R组拔出线栓恢复血液灌注，HT组和NT组分别通过颈动脉灌注4 ℃生理盐水和37 ℃常温盐水，然后恢复血液灌注。采用Zea Longa 5 分制评分评估大鼠神经功能，TTC染色测定脑梗死面积，采用TUNEL染色测定皮质缺血半暗带细胞凋亡率，Western blot 测定线粒体分裂蛋白1及cyto-Cyto c 的表达，qRT-PCR测定Fis1 mRNA的表达，电镜下观察线粒体超微结构。结果显示：(1)与sham组相比，其他3组再灌注6，24，48 h大鼠皮质缺血半暗带细胞凋亡率，线粒体分裂蛋白1蛋白及其mRNA的表达，cyto-Cyto c蛋白表达均增加，再灌注24 h脑梗死面积增加；(2)HT组可对抗I/R引起的上述变化，而NT组则不能；(3)上述数据说明，选择性脑低温可以抑制线粒体分裂蛋白1的表达，减少细胞凋亡，进而减轻大鼠局部脑I/R损伤。

orcid: 0000-0002-8938-8851 (Ming-Shan Wang)

关键词: 卒中, 缺血再灌注损伤, 神经保护, 低温, 选择性脑低温, Fis1, 线粒体, 线粒体分裂, 线粒体超微结构, 凋亡

Abstract: Selective brain hypothermia is considered an effective treatment for neuronal injury after stroke, and avoids the complications of general hypothermia. However, the mechanisms by which selective brain hypothermia affects mitochondrial fission remain unknown. In this study, we investigated the effect of selective brain hypothermia on the expression of fission 1 (Fis1) protein, a key factor in the mitochondrial fission system, during focal cerebral ischemia/reperfusion injury. Sprague-Dawley rats were divided into four groups. In the sham group, the carotid arteries were exposed only. In the other three groups, middle cerebral artery occlusion was performed using the intraluminal filament technique. After 2 hours of occlusion, the filament was slowly removed to allow blood reperfusion in the ischemia/reperfusion group. Saline, at 4°C and 37°C, were perfused through the carotid artery in the hypothermia and normothermia groups, respectively, followed by restoration of blood flow. Neurological function was assessed with the Zea Longa 5-point scoring method. Cerebral infarct volume was assessed by 2,3,5-triphenyltetrazolium chloride staining, and apoptosis was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling staining. Fis1 and cytosolic cytochrome c levels were assessed by western blot assay. Fis1 mRNA expression was assessed by quantitative reverse transcription-polymerase chain reaction. Mitochondrial ultrastructure was evaluated by transmission electron microscopy. Compared with the sham group, apoptosis, Fis1 protein and mRNA expression and cytosolic cytochrome c levels in the cortical ischemic penumbra and cerebral infarct volume were increased after reperfusion in the other three groups. These changes caused by cerebral ischemia/reperfusion were inhibited in the hypothermia group compared with the normothermia group. These findings show that selective brain hypothermia inhibits Fis1 expression and reduces apoptosis, thereby ameliorating focal cerebral ischemia/reperfusion injury in rats. Experiments were authorized by the Ethics Committee of Qingdao Municipal Hospital of China (approval No. 2019008).

Key words: apoptosis, Fis1, hypothermia, ischemia/reperfusion injury, mitochondria, mitochondrial fission, mitochondrial ultrastructure, neuroprotection, selective brain hypothermia, stroke