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 (11): 2003-2010.doi: 10.4103/1673-5374.253174

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

深低温停循环大脑中微小RNA的表达变化

  

  • 出版日期:2019-11-15 发布日期:2019-11-15
  • 基金资助:

    中国国家自然科学基金项目（81401084），北京市医院管理局启航计划（DFL20150802），北京215高级医疗卫生人才学术带头人计划（008-0027），北京市卫健委项目（PXM2017_026283_000002），北京市医院管理局临床医学发展专项资助支持（ZYLX201706，303-01-005-0137-11）

MicroRNA expression in the hippocampal CA1 region under deep hypothermic circulatory arrest

Xiao-Hua Wang 1, 2, 3 , Dong-Xu Yao 1, 2, 3 , Xiu-Shu Luan 1, 2, 3 , Yu Wang 1, 2, 3 , Hai-Xia Liu 1, 2, 3 , Bei Liu 1, 2, 3 , Yang Liu 1, 2, 3 , Lei Zhao 1, 2, 3 , Xun-Ming Ji 4, 5 , Tian-Long Wang 1, 2, 3   

  1. 1 Department of Anesthesiology, Xuanwu Hospital, Capital Medical University, Beijing, China
    2 Institute of Geriatrics, Beijing, China
    3 National Clinical Research Center for Geriatric Disorders, Beijing, China
    4 Department of Neurosurgery, Xuanwu Hospital, Capital Medical University, Beijing, China
    5 Cerebrovascular Research Center, Xuanwu Hospital, Capital Medical University, Beijing, China
  • Online:2019-11-15 Published:2019-11-15
  • Contact: Tian-Long Wang, MD, w_tl5595@yahoo.com; Xun-Ming Ji, jixunming@vip.163.com.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 81401084 (to XHW); the Beijing Municipal Administration of Hospital Ascent Plan in China, No. DFL20150802 (to TLW); the Beijing 215 High Level Healthcare Talent Plan Academic Leader in China, No. 008-0027 (to TLW); the Beijing Municipal Commission of Health and Family Planning in China, No. PXM2017_026283_000002 (to TLW); the Beijing Municipal Administration of Hospitals Clinical Medicine Development of Special Funding Support in China, No. ZYLX201706 (to TLW), 303-01-005-0137-11 (to TLW), 65683.00 (to TLW).

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摘要：

在深低温停循环进行胸主动脉以及复杂心脏病手术的同时，尚未深刻认为深低温停循环期间的脑保护作用机制。为此，实验设计对新生仔猪进行体外循环实施为期为60min的14°C深低温停循环。(1)使用微阵列扫描分析海马中miRNA的表达情况，TargetScan 6.2，RNA22 v2.0，miRWalk 2.0和miRanda进行潜在目标miRNA预测，以基因本体富集分析发现相关功能通路，以实时定量逆转录PCR对相关miRNA的变化进行验证；(2)结果显示，深低温停循环后仔猪海马中有35种miRNA的表达发生了变化，其中22种表达下调，13种表达上调；差异表达的miRNA参与神经元投射（CDK16和SLC1A2）、中枢神经系统发育（FOXO3，TYRO3和SLC1A2）、离子跨膜转运蛋白活性（iATP2B2和SLC1A2）以及白细胞介素6受体结合（IL6R），且这些均为深低温停循环期间参与脑保护的关键信号通路。同时实时定量反转录PCR证实了微阵列分析得出的结果；(3)实验结果阐明了深低温停循环中转录调控的新作用，为利用miRNA开发治疗脑损伤方法可提供有意义的帮助；(4)实验已于2017-03-01经首都医科大学宣武医院动物伦理委员会批准，批准号XW-INI-AD2017-0112。

orcid: 0000-0003-1636-0142(Tian-Long Wang)

关键词: 脑保护, 深低温停循环, 基因本体富集分析, 微小RNA, 海马, 转录后表达, 基因芯片, 生物信息学, 神经再生

Abstract:

Using deep hypothermic circulatory arrest, thoracic aorta diseases and complex heart diseases can be subjected to corrective procedures. However, mechanisms underlying brain protection during deep hypothermic circulatory arrest are unclear. After piglet models underwent 60 minutes of deep hypothermic circulatory arrest at 14°C, expression of microRNAs (miRNAs) was analyzed in the hippocampus by microarray. Subsequently, TargetScan 6.2, RNA22 v2.0, miRWalk 2.0, and miRanda were used to predict potential targets, and gene ontology enrichment analysis was carried out to identify functional pathways involved. Quantitative reverse transcription-polymerase chain reaction was conducted to verify miRNA changes. Deep hypothermic circulatory arrest altered the expression of 35 miRNAs. Twenty-two miRNAs were significantly downregulated and thirteen miRNAs were significantly upregulated in the hippocampus after deep hypothermic circulatory arrest. Six out of eight targets among the differentially expressed miRNAs were enriched for neuronal projection (cyclin dependent kinase, CDK16 and SLC1A2), central nervous system development (FOXO3, TYRO3, and SLC1A2), ion transmembrane transporter activity (ATP2B2 and SLC1A2), and interleukin-6 receptor binding (IL6R) – these are the key functional pathways involved in cerebral protection during deep hypothermic circulatory arrest. Quantitative reverse transcription-polymerase chain reaction confirmed the results of microarray analysis. Our experimental results illustrate a new role for transcriptional regulation in deep hypothermic circulatory arrest, and provide significant insight for the development of miRNAs to treat brain injuries. All procedures were approved by the Animal Care Committee of Xuanwu Hospital, Capital Medical University, China on March 1, 2017 (approval No. XW-INI-AD2017-0112).

Key words: nerve regeneration, cerebral protection, deep hypothermic circulatory arrest, gene ontology enrichment analysis, microRNA, hippocampus, post-transcriptional expression, microarray, bioinformatics, neural regeneration