中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2017, Vol. 12 ›› Issue (12): 2007-2013.doi: 10.4103/1673-5374.221157

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

正常体温条件下窒息致心脏骤停早期大鼠海马神经元损伤与肿瘤坏死因子α的免疫反应

  

  • 收稿日期:2017-11-13 出版日期:2017-12-15 发布日期:2017-12-15
  • 基金资助:

    韩国教育部国家研究基金；韩国科学，ICT和未来计划项目

Neuronal injury and tumor necrosis factor-alpha immunoreactivity in the rat hippocampus in the early period of asphyxia-induced cardiac arrest under normothermia

Hyun-Jin Tae1, Il Jun Kang2, Tae-Kyeong Lee3, Jeong Hwi Cho3, Jae-Chul Lee3, Myoung Cheol Shin4, Yoon Sung Kim4, 5, Jun Hwi Cho4,Jong-Dai Kim6, Ji Hyeon Ahn7, Joon Ha Park7, In-Shik Kim1, Hyang-Ah Lee8, Yang Hee Kim9, Moo-Ho Won3, Young Joo Lee10   

  1. 1 Bio-Safety Research Institute, College of Veterinary Medicine, Chonbuk National University, Iksan, South Korea
    2 Department of Food Science and Nutrition, Hallym University, Chuncheon, South Korea
    3 Department of Neurobiology, School of Medicine, Kangwon National University, Chuncheon, South Korea
    4 Department of Emergency Medicine, School of Medicine, Kangwon National University, Chuncheon, South Korea
    5 Department of Emergency Medicine, Samcheok Medical Center, Samcheok, South Korea
    6 Division of Food Biotechnology, School of Biotechnology, Kangwon National University, Chuncheon, South Korea
    7 Department of Biomedical Science, Research Institute of Bioscience and Biotechnology, Hallym University, Chuncheon, South Korea
    8 Department of Obstetrics and Gynecology, School of Medicine, Kangwon National University, Chuncheon, South Korea
    9 Department of Surgery, School of Medicine, Kangwon National University, Chuncheon, South Korea
    10 Department of Emergency Medicine, Seoul Hospital, College of Medicine, Sooncheonhyang University, Seoul, South Korea
  • Received:2017-11-13 Online:2017-12-15 Published:2017-12-15
  • Contact: Moo-Ho Won, D.V.M., Ph.D.or Young Joo Lee, M.D., Ph.D.,mhwon@kangwon.ac.kr or brugada@naver.com.
  • Supported by:

    This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF),the Ministry of Education (NRF-2014R1A1A2057263), by the Basic Science Research Program through the National Research Foundation of Korea(NRF) funded by the Ministry of Science, ICT & Future Planning (NRF-2017R1A2B4009079 & NRF-2017R1A2B4008403), and by the Bio-Synergy Research Project (NRF-2015M3A9C4076322) of the Ministry of Science, ICT and Future Planning through the National Research Foundation.

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摘要：

首次经历心脏骤停（CA）后恢复自主循环的患者存活率低。实验以成年雄性Sprague-Daley大鼠诱导窒息致心脏骤停模型，并维持体温为(37 ± 0.5)°C，然后在心肺复苏后观察存活率；以焦油紫（CV）和Fluore-Jade B（FJ B）染色方法检测海马神经元损伤和退变情况；应用免疫组化染色法检测海马小胶质细胞标志物离子化的钙结合接头分子1（Iba-1），星形胶质细胞标志物胶质纤维酸性蛋白（GFAP）和炎症因子肿瘤坏死因子免疫反应，以反映正常体温条件下窒息致心脏骤停引起的促炎症反应。CA模型大鼠在造模后6h，1和8d生存率逐渐下降，分别为63%，37%，8%。直到心脏骤停后1d未在海马中发现有神经元损伤，但在心脏骤停后2d，海马CA区锥体细胞层中的一些神经元死亡。CA1区的Iba-1免疫反应阳性小胶质细胞直到心脏骤停后1d才发生变化，并在心脏骤停后2d在所有层中激活，表现为有短而粗突起的增大细胞体）。心脏骤停后2 d，胶质纤维酸性蛋白免疫阳性星形胶质细胞变化不明显；心脏骤停后6h，CA1区锥体细胞层神经元中肿瘤坏死因子α免疫反应明显下降，且下降趋势持续至心脏骤停后1d，心脏骤停后2d又再次明显升高。说明心脏骤停早期诱导体温正常大鼠死亡率高与肿瘤坏死因子α表达的早期升高有关，而非海马CA1区神经元损伤。

orcid:0000-0002-7178-6501(Moo-Ho Won)
0000-0003-2412-270X(Young Joo Lee)

关键词: 神经再生, 心脏骤停后综合征, 体温正常, 神经元损伤, 胶质细胞增生, 肿瘤坏死因子α

Abstract:

Low survival rate occurs in patients who initially experience a spontaneous return of circulation after cardiac arrest (CA). In this study, we induced asphyxial CA in adult male Sprague-Daley rats, maintained their body temperature at 37 ± 0.5°C, and then observed the survival rate during the post-resuscitation phase. We examined neuronal damage in the hippocampus using cresyl violet (CV) and Fluore-Jade B (F-J B) staining, and pro-inflammatory response using ionized calcium-binding adapter molecule 1 (Iba-1), glial fibrillary acidic protein (GFAP), and tumor necrosis factor-alpha (TNF-α) immunohistochemistry in the hippocampus after asphyxial CA in rats under normothermia. Our results show that the survival rate decreased gradually post-CA (about 63% at 6 hours, 37% at 1 day, and 8% at 2 days post-CA). Rats were sacrificed at these points in time post-CA, and no neuronal damage was found in the hippocampus until 1 day post-CA. However, some neurons in the stratum pyramidale of the CA region in the hippocampus were dead 2 days post-CA. Iba-1 immunoreactive microglia in the CA1 region did not change until 1 day post-CA, and they were activated (enlarged cell bodies with short and thicken processes) in all layers 2 days post-CA. Meanwhile, GFAP-immunoreactive astrocytes did not change significantly until 2 days post-CA. TNF-αimmunoreactivity decreased significantly in neurons of the stratum pyramidale in the CA1 region 6 hours post-CA, decreased gradually until 1 day post-CA, and increased significantly again 2 days post-CA. These findings suggest that low survival rate of normothermic rats in the early period of asphyxia-induced CA is related to increased TNF-α immunoreactivity, but not to neuronal damage in the hippocampal CA1 region.

Key words: nerve regeneration, post-cardiac arrest syndrome, normothermia, neuronal damage, gliosis, tumor necrosis factor-alpha, neural regeneration