中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2016, Vol. 11 ›› Issue (11): 1824-1829.doi: 10.4103/1673-5374.194754

• 原著:脊髓损伤修复保护与再生 • 上一篇    下一篇

缺血再灌注损伤脊髓组织不同时序变化的起作用的关键基因

  

  • 出版日期:2016-11-30 发布日期:2016-11-30
  • 基金资助:
    国家自然科学基金（81350013, 31572217, 81672263）

Key genes expressed in different stages of spinal cord ischemia/reperfusion injury

Jian-an Li, Chun-fang Zan, Peng Xia, Chang-jun Zheng, Zhi-ping Qi, Chun-xu Li, Zhi-gang Liu, Ting-ting Hou*, Xiao-yu Yang*   

  1. Department of Orthopaedics, the Second Hospital of Jilin University, Changchun, Jilin Province, China
  • Online:2016-11-30 Published:2016-11-30
  • Contact: Ting-ting Hou, Ph.D., or Xiao-yu Yang, M.D., httandtina@163.com or yangxiaoyu88@sina.com.
  • Supported by:
    This study was supported by a Grant from the National Natural Science Foundation of China, No. 81350013, 31572217, and 81672263.

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摘要：

脊髓缺血再灌注损伤后microRNA不同时序的变化目前尚不明确。实验设计夹闭腹主动脉法制作脊髓缺血再灌注损伤24，48 h大鼠模型，同时设单纯缺血组为对照，拟说明问题。切取各组缺血脊髓节段，其中一部分组织进行苏木精-伊红染色，光学显微镜下可见神经元损伤和组织水肿状况在缺血90 min再灌注24 h达到顶峰，其后于缺血90 min再灌注48 h有所缓解。另一部分组织提取RNA做扩增、标记和芯片杂交，获得microRNA的表达谱，通过整合分析筛选出4种microRNA：miR-22-3p、miR-743b-3p、miR-201-5p and miR-144-5p及其靶基因(Tmem69、Cxcl10等)，见3组中rno-miR-22-3p持续表达上调，且在单纯缺血组中上调倍数最高；但rno-miR-743b-3p、rno-miR-201-5p 和rno-miR-144-5p在3组中表达下调；实验还同时构建了microRNA与靶基因关系网络图。结果显示，实验成功筛选了脊髓缺血再灌注损伤不同时间段与演进过程中起主导作用的关键基因，可成为脊髓损伤机制研究的参考依据。 

orcid: 0000-0001-9010-2147 (Ting-ting Hou), 0000-0001-9388-3794 (Xiao-yu Yang)

关键词: 神经再生, 脊髓损伤, 缺血再灌注损伤, mRNA, MicroRNA, 生物信息学, Tmem69基因, Cxcl10基因, 神经元再生, 转录组学, MicroRNA序列

Abstract: The temporal expression of microRNA afer spinal cord ischemia/reperfusion injury is not yet fully understood. In the present study, we established a model of spinal cord ischemia in Sprague-Dawley rats by clamping the abdominal aorta for 90 minutes, before allowing reperfusion for 24 or 48 hours. A sham-operated group underwent surgery but the aorta was not clamped. Te damaged spinal cord was removed for hematoxylin-eosin staining and RNA extraction. Neuronal degeneration and tissue edema were the most severe in the 24- hour reperfusion group, and milder in the 48-hour reperfusion group. RNA amplifcation, labeling, and hybridization were used to obtain the microRNA expression profles of each group. Bioinformatics analysis confrmed four differentially expressed microRNAs (miR-22-3p, miR-743b-3p, miR-201-5p and miR-144-5p) and their common target genes (Tmem69 and Cxcl10). Compared with the sham group, miR- 22-3p was continuously upregulated in all three ischemia groups but was highest in the group with no reperfusion, whereas miR-743b-3p, miR-201-5p and miR-144-5p were downregulated in the three ischemia groups. We have successfully identifed the key genes expressed at different stages of spinal cord ischemia/reperfusion injury, which provide a reference for future investigations into the mechanism of spinal cord injury.

Key words: nerve regeneration, spinal cord injury, ischemia/reperfusion injury, mRNA, microRNA, bioinformatics, Tmem69, Cxcl10, transcriptome, microRNA arrays, neural regeneration