中国神经再生研究(英文版) ›› 2020, Vol. 15 ›› Issue (6): 1102-1110.doi: 10.4103/1673-5374.270418

• 原著:脊髓损伤修复保护与再生 • 上一篇    下一篇

脊髓损伤模型小鼠MicroRNA-21-5p的比较蛋白质组学变化及在不同途径中的可能作用

  

  • 出版日期:2020-06-15 发布日期:2020-07-02

Comparative proteomes change and possible role in different pathways of microRNA-21a-5p in a mouse model of spinal cord injury

Almaghalsa-Ziad Mohammed, Hong-Xia Du, Hong-Liang Song, Wei-Ming Gong, Bin Ning, Tang-Hong Jia   

  1. Department of Spinal Surgery, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong Province, China
  • Online:2020-06-15 Published:2020-07-02
  • Contact: Bin Ning, ningbin@sdu.edu.cn.

摘要:

课题组以往研究发现,microRNA21a-5p(miR-21a-5p)基因敲除可改善脊髓损伤模型小鼠脊髓损伤后运动功能的恢复,但具体分子机制尚不清楚。为此,实验采用重物打击法建立脊髓损伤小鼠模型,利用蛋白质组学方法来观察miR-21a-5p基因敲除的脊髓损伤模型小鼠脊髓组织中不同蛋白质的表达,并设未经基因敲除的脊髓损伤模型小鼠为阴性对照组。对比发现:(1)在miR-21a-5p基因敲除后,与NC组相比,基因敲除组小鼠在炎症调节,细胞氧化应激保护,细胞氧化还原稳态和细胞维持中发挥重要作用的多数蛋白质表达均上调;(2)KEGG途径分析发现,两组小鼠均发现了KEGG相关的富集途径,如氧化磷酸化途径,其与帕金森病,亨廷顿病,阿尔茨海默病和心肌收缩相关;(3)实验还发现miR-21a-5p可能是肌萎缩侧索硬化途径中的潜在生物标志物,因为miR-21a-5p在该途径中表达下调;(4)上述数据说明,实验成功检测出了在脊髓损伤中发挥潜在作用的一些重要蛋白质,研究这些蛋白质及其在脊髓损伤恢复中的相互关系,将为未来脊髓损伤生物标志物的研究提供参考依据。所有实验程序和方案均经中国山东大学实验动物伦理委员会批准。

orcid: 0000-0002-7592-9485 (Bin Ning)

关键词: 脊髓损伤, microRNA, 蛋白质组学, 生物标志物, 炎症, 小鼠, stathmin, 生物信息学, 线粒体, 途径分析

Abstract: Our previous study found that microRNA-21a-5p (miR-21a-5p) knockdown could improve the recovery of motor function after spinal cord injury in a mouse model, but the precise molecular mechanism remains poorly understood. In this study, a modified Allen’s weight drop was used to establish a mouse model of spinal cord injury. A proteomics approach was used to understand the role of differential protein expression with miR-21a-5p knockdown, using a mouse model of spinal cord injury without gene knockout as a negative control group. We found that after introducing miR-21a-5p knockdown, proteins that played an essential role in the regulation of inflammatory processes, cell protection against oxidative stress, cell redox homeostasis, and cell maintenance were upregulated compared with the nega- tive control group. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis identified enriched pathways in both groups, such as the oxidative phosphorylation pathway, which is relevant to Parkinson’s disease, Huntington’s disease, Alzheimer’s disease, and cardiac muscle contraction. We also found that miR-21a-5p could be a potential biomarker for amyotrophic lateral sclerosis, as miR-21a- 5p becomes deregulated in this pathway. These results indicate successful detection of some important proteins that play potential roles in spinal cord injury. Elucidating the relationship between these proteins and the recovery of spinal cord injury will provide a reference for future research of spinal cord injury biomarkers. All experimental procedures and protocols were approved by the Experimental Animal Ethics Committee of Shandong University of China on March 5, 2014.

Key words: bioinformatics, biomarker, inflammation, microRNA, mitochondria, mouse, pathway analysis, proteomics, spinal cord injury,
stathmin