中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2019, Vol. 14 ›› Issue (8): 1394-1403.doi: 10.4103/1673-5374.253524

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

轻重度或致死性短暂性全脑缺血沙鼠海马神经元丢失和神经胶质增生的时程模式

  

  • 出版日期:2019-08-15 发布日期:2019-08-15
  • 基金资助:

    基础科学研究计划通过由教育部资助的韩国国家研究基金会（NRF）（NRF-2016R1D1A1B01011790; JHC）支持; 韩国科学，ICT和未来规划部国家研究基金会（NRF）资助的基础科学研究计划（NRF-2017R1A2B4009079; MHW）和农业科学技术发展合作研究计划（PJ01329401）

Time-course pattern of neuronal loss and gliosis in gerbil hippocampi following mild, severe, or lethal transient global cerebral ischemia

Tae-Kyeong Lee 1 , Hyunjung Kim 1 , Minah Song 1 , Jae-Chul Lee 1 , Joon Ha Park 2 , Ji Hyeon Ahn 2 , Go Eun Yang 3 , Hyeyoung Kim 4, 5 , Taek Geun Ohk 5 , Myoung Cheol Shin 5 , Jun Hwi Cho 5 , Moo-Ho Won 1   

  1. 1 Department of Neurobiology, School of Medicine, Kangwon National University, Chuncheon, Gangwon, Republic of Korea
    2 Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University, Chuncheon, Gangwon, Republic of Korea
    3 Department of Radiology, Kangwon National University Hospital, Chuncheon, Gangwon, Republic of Korea
    4 Department of Anesthesiology and Pain Medicine, Chungju Hospital, Konkuk University School of Medicine, Chungju Chungcheongbuk, Republic of Korea
    5 Department of Emergency Medicine, School of Medicine, Kangwon National University, Chuncheon, Gangwon, Republic of Korea
  • Online:2019-08-15 Published:2019-08-15
  • Contact: Moo-Ho Won, DVM, PhD, mhwon@kangwon.ac.kr; Jun Hwi Cho, MD, PhD, cjhemd@kangwon.ac.kr.
  • Supported by:

    This study was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2016R1D1A1B01011790; to JHC); Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT & Future Planning (NRF-2017R1A2B4009079; to MHW), and by Cooperative Research Program for Agriculture Science and Technology Development (Project No. PJ01329401; to MHW) Rural Development Administration, Republic of Korea.

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摘要：

全脑短暂性缺血（TI）在脆弱的大脑区域中可引起神经元丢失/死亡，TI后纹状体、新皮质和海马神经元可变松散，仅仅5min的TI可以在缺血后4d引起海马CA1区锥体神经元死亡。为了解5min（轻度），15min（严重）和20min（致死）双侧颈动脉闭塞（BCCAO）对沙鼠行为改变及海马CA1-3区和齿状回神经元死亡、星形细胞和小胶质细胞增生的影响，实验以自发运动活动（SMA）测试评估沙鼠行为学变化，以甲酚紫染色检测细胞分布，以神经元核免疫组化染色检测神经元分布，以Fluoro-Jade B组织荧光染色检测神经元死亡。分别通过胶质纤维酸性蛋白（GFAP）和离子化钙结合衔接分子1（Iba1）免疫组织化学染色检测星形细胞增生和小胶质细胞增生。首先，实验发现BCCAO 20min沙鼠在至少2d内死亡。缺血15min的沙鼠在术后2d诱发CA1-3锥体细胞死亡，5天后齿状回颗粒细胞死亡。在BCCAO5min的沙鼠中未发现这种效应。同时，BCCAO15min沙鼠胶质细胞增生比在BCCAO5分钟沙鼠更快且更严重地进展。实验结果表明TI后海马中神经元丢失情况依脑区不同和缺血严重程度而显著不同。

orcid: 0000-0002-7178-6501 (Moo-Ho Won)
           0000-0001-8558-3564 (Jun Hwi Cho)

关键词: 短暂性全脑缺血, 迟发性神经元死亡, 胶质激活, 缺血持续时间, 海马, 自发运动, 蒙古沙鼠, 组织学, 神经再生

Abstract:

Transient ischemia in the whole brain leads to neuronal loss/death in vulnerable brain regions. The striatum, neocortex and hippocampus selectively loose specific neurons after transient ischemia. Just 5 minutes of transient ischemia can cause pyramidal neuronal death in the hippocampal cornu ammonis (CA) 1 field at 4 days after transient ischemia. In this study, we investigated the effects of 5-minute (mild), 15-minute (severe), and 20-minute (lethal) transient ischemia by bilateral common carotid artery occlusion (BCCAO) on behavioral change and neuronal death and gliosis (astrocytosis and microgliosis) in gerbil hippocampal subregions (CA1–3 region and dentate gyrus). We performed spontaneous motor activity test to evaluate gerbil locomotor activity, cresyl violet staining to detect cellular distribution, neuronal nuclei immunohistochemistry to detect neuronal distribution, and Fluoro-Jade B histofluorescence to evaluate neuronal death. We also conducted immunohistochemical staining for glial fibrillary acidic protein and ionized calcium-binding adapter molecule 1 (Iba1) to evaluate astrocytosis and microgliosis, respectively. Animals subjected to 20-minute BCCAO died in at least 2 days. BCCAO for 15 minutes led to pyramidal cell death in hippocampal CA1–3 region 2 days later and granule cell death in hippocampal dentate gyrus 5 days later. Similar results were not found in animals subjected to 5-minute BCCAO. Gliosis was much more rapidly and severely progressed in animals subjected to 15-minute BCCAO than in those subjected to 5-minute BCCAO. Our results indicate that neuronal loss in the hippocampal formation following transient ischemia is significantly different according to regions and severity of transient ischemia. The experimental protocol was approved by Institutional Animal Care and Use Committee (AICUC) of Kangwon National University (approval No., KW-180124-1) on May 22, 2018.

Key words: transient global brain ischemia, delayed neuronal death, glial activation, ischemic duration, hippocampus, spontaneous motor activity, Mongolian gerbil, histology, neural regeneration