中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2018, Vol. 13 ›› Issue (1): 86-93.doi: 10.4103/1673-5374.224373

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

电针预处理保护局灶性脑缺血再灌注损伤：与抑制动力相关蛋白1有关

  

  • 收稿日期:2018-01-02 出版日期:2018-01-15 发布日期:2018-01-15
  • 基金资助:

      山东省自然科学基金项目（ZR2015HM023）青岛市市南区科技项目（2016-3-029-YY）

Electroacupuncture preconditioning protects against focal cerebral ischemia/reperfusion injury via suppression of dynamin-related protein 1

Gao-feng Zhang1, Pei Yang2, Zeng Yin1, Huai-long Chen1, Fu-guo Ma1, Bin Wang1, Li-xin Sun1, Yan-lin Bi1, Fei Shi1, Ming-shan Wang1   

  1. 1 Department of Anesthesiology, Affiliated Qingdao Municipal Hospital of Qingdao University, Qingdao, Shandong Province, China
    2 Department of Public Health, Affiliated Qingdao Municipal Hospital of Qingdao University, Qingdao, Shandong Province, China
  • Received:2018-01-02 Online:2018-01-15 Published:2018-01-15
  • Contact: Ming-shan Wang,wmhill@163.com.
  • Supported by:

    This study was supported by the Natural Science Foundation of Shandong Province of China, No. ZR2015HM023; a grant from the Science and Technology Plan Project of Shinan District of Qingdao City of China, No. 2016-3-029-YY.

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摘要：

电针百会（GV20）预处理可以减轻局灶性脑缺血再灌注损伤，但其确切机制仍不明确。由于脑缺血再灌注损伤后，动力相关蛋白1介导的线粒体裂变会造成神经元凋亡。作者假设电针预处理可能抑制动力相关蛋白1表达，进而干预线粒体裂变产生脑保护作用。实验以电针百会（深度2mm、强度1 mA、频率2/15 Hz，持续30min，1次/d）连续5d预处理后24h，建立大脑中动脉闭塞局灶性脑缺血再灌注损伤大鼠模型。以Longa神经缺陷评分评价神经功能，以苏木精-伊红染色观察损伤侧缺血半暗带病理变化，以TUNEL染色检测损伤侧缺血半暗带中细胞的凋亡，以透射电镜观察损伤侧缺血半暗带线粒体超微结构，以Western blot检测损伤侧缺血半暗带动力相关蛋白1和细胞色素C表达水平。结果显示，电针预处理降低了局灶性脑缺血再灌注损伤大鼠损伤侧缺血半暗带中总动力相关蛋白1以及线粒体内动力相关蛋白1水平，降低了损伤侧缺血半暗带中总细胞色素C及细胞色素C的水平，较好的维持了损伤侧缺血半暗带中线粒体形态，减少了损伤侧缺血半暗带中凋亡细胞比例，从而显著改善了神经功能。表明电针预处理对局灶性脑缺血再灌注损伤的神经保护作用与抑制动力相关蛋白1有确切关系。

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

关键词: 神经再生, 电针, 局灶性脑缺血再灌注损伤, 动力相关蛋白1, 死亡相关蛋白激酶, 线粒体动力学, 线粒体超微结构, 细胞凋亡, 细胞色素C

Abstract:

Electroacupuncture preconditioning at acupoint Baihui (GV20) can reduce focal cerebral ischemia/reperfusion injury. However, the precise protective mechanism remains unknown. Mitochondrial fission mediated by dynamin-related protein 1 (Drp1) can trigger neuronal apoptosis following cerebral ischemia/reperfusion injury. Herein, we examined the hypothesis that electroacupuncture pretreatment can regulate Drp1, and thus inhibit mitochondrial fission to provide cerebral protection. Rat models of focal cerebral ischemia/reperfusion injury were established by middle cerebral artery occlusion at 24 hours after 5 consecutive days of preconditioning with electroacupuncture at GV20 (depth 2 mm, intensity 1 mA, frequency 2/15 Hz, for 30 minutes, once a day). Neurological function was assessed using the Longa neurological deficit score. Pathological changes in the ischemic penumbra on the injury side were assessed by hematoxylin-eosin staining. Cellular apoptosis in the ischemic penumbra on the injury side was assessed by terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling staining. Mitochondrial ultrastructure in the ischemic penumbra on the injury side was assessed by transmission electron microscopy. Drp1 and cytochrome c expression in the ischemic penumbra on the injury side were assessed by western blot assay. Results showed that electroacupuncture preconditioning decreased expression of total and mitochondrial Drp1, decreased expression of total and cytosolic cytochrome c, maintained mitochondrial morphology and reduced the proportion of apoptotic cells in the ischemic penumbra on the injury side, with associated improvements in neurological function. These data suggest that electroacupuncture preconditioning-induced neuronal protection involves inhibition of the expression and translocation of Drp1.

Key words: nerve regeneration, electroacupuncture, focal cerebral ischemia/reperfusion injury, dynamin-related protein 1, death-associated protein kinases, mitochondrial dynamics, mitochondrial ultrastructure, apoptosis, cytochrome c, neural regeneration