中国组织工程研究 ›› 2018, Vol. 22 ›› Issue (32): 5180-5185.doi: 10.3969/j.issn.2095-4344.0773

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

抑制miR-21a-5p能促进脊髓损伤小鼠运动功能的恢复

王文朝,苏延林,李洪飞,沈  霖,陈佳男,潘信达,贾堂宏   

  1. 山东大学附属济南市中心医院骨外科,山东省济南市  250013
  • 收稿日期:2018-01-27 出版日期:2018-11-18 发布日期:2018-11-18
  • 通讯作者: 贾堂宏,教授,博士生导师,主任医师,山东大学附属济南市中心医院骨外科,山东省济南市 250013
  • 作者简介:王文朝,男,1991年生,山东省济宁市人,汉族,山东大学骨外科在读硕士,主要从脊髓损伤、非编码RNA方面的研究。
  • 基金资助:

    国家自然科学基金青年科学基金项目(81401014);国家自然科学基金面上项目(81771346);中国博士后科学基金面上项目(2014M561935);中国博士后科学基金特别资助项目(2015T80725)

miR-21a-5p knockdown promotes the locomotor function recovery of mouse models of spinal cord injury

Wang Wen-zhao, Su Yan-lin, Li Hong-fei, Shen Lin, Chen Jia-nan, Pan Xin-da, Jia Tang-hong   

  1. Department of Orthopedics, Jinan Central Hospital Affiliated to Shandong University, Jinan 250013, Shandong Province, China
  • Received:2018-01-27 Online:2018-11-18 Published:2018-11-18
  • Contact: Jia Tang-hong, Professor, Doctoral supervisor, Chief physician, Department of Orthopedics, Jinan Central Hospital Affiliated to Shandong University, Jinan 250013, Shandong Province, China
  • About author:Wang Wen-zhao, Master candidate, Department of Orthopedics, Jinan Central Hospital Affiliated to Shandong University, Jinan 250013, Shandong Province, China
  • Supported by:

    the National Natural Science Foundation of China for the Youth, No. 81401014; the National Natural Science Foundation of China, No. 81771346; the General Project of Postdoctoral Science Foundation of China, No. 2014M561935; the Special Project of Postdoctoral Science Foundation of China, No. 2015T80725

摘要:

文章快速阅读:

文题释义:
MicroRNAs(miRNAs):是在真核生物中发现的一类内源性的具有调控功能的非编码RNA,其长度为20-25个核苷酸。成熟的miRNAs是由较长的初级转录本(pri-miRNA)经过一系列核酸酶的剪切加工而产生的,随后组装进RNA诱导的沉默复合体,通过碱基互补配对的方式识别靶mRNA,并根据互补程度的不同指导沉默复合体降解靶mRNA或者阻遏靶mRNA的翻译。近年来的研究表明miRNA参与了多种调节途径,包括发育、病毒防御、造血过程、器官形成、细胞增殖和凋亡、脂肪代谢等等。
纤维连接蛋白(fibronectin,FN):为一族高分子糖蛋白,由间叶细胞分泌,广泛分布于人体细胞表面和血浆中,有促进巨噬细胞的吞噬功能,促进细胞与纤维基质间连接的生理作用。纤连蛋白与机体创伤修复、组织炎症、纤维化及硬化过程等有密切关系。纤连蛋白也是一种重要的调理素,对严重疾病的诊断有重要价值。
摘要
背景
:脊髓损伤后病理生理改变复杂,尚无有效的治疗手段。miRNAs作为可以精细调控多种基因表达的内源性非编码RNA,可为脊髓损伤的治疗提供新思路。
目的:探究抑制miR-21a-5p对小鼠脊髓损伤后运动功能恢复的影响及可能的机制。
方法:第1步:Allen’s打击法构建小鼠脊髓损伤模型,提取术后3 d损伤处脊髓组织(n=24)对比假手术组(n=24)筛选出差异表达miRNA;第2步:脊髓损伤后鞘内注射miRNA抑制剂(antagomir-21a)构建观察组(n=24),对照组造模成功后鞘内注射等量的antagomir做对照(n=24)、假手术组(n=24)术后不做特殊处理;对比各组行为学评分(BMS评分),检测脊髓损伤相关标志物的表达。
结果与结论:①小鼠脊髓损伤后miR-21a-5p的表达较假手术组显著升高(P < 0.05);②观察组小鼠脊髓损伤后抑制miR-21a-5p,假手术组与脊髓损伤组小鼠运动功能评分差异明显(P < 0.05),术后7 d观察组小鼠运动功能评分逐渐超过对照组,术后14 d显著高于对照组(P < 0.05);③观察组小鼠脊髓组织中纤维连接蛋白与CSPGs的表达量低于对照组,且神经营养因子的表达高于对照组(P < 0.05);④结果说明:小鼠脊髓损伤后miR-21a-5p的表达上调,抑制miR-21a-5p的表达可以抑制瘢痕形成并增加神经营养因子的分泌,从而促进运动功能的恢复。

中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程
ORCID: 0000-0001-5106-4862(王文朝)

关键词: mRNAs, miR-21a-5p, 脊髓损伤, 瘢痕形成, 运动功能, 神经功能, antagomir-21a, BMS评分, 神经营养因子, 组织构建

Abstract:

BACKGROUND: The pathological changes following spinal cord injury (SCI) are complex, and there is a lack of effective treatment method. miRNAs are non-coding RNA molecules that post-transcriptionally regulate gene expression, which provides a novel treatment strategy for SCI.
OBJECTIVE: To investigate the effect of miR-21a-5p on the locomotor function post-SCI in mice and explore the related mechanisms.
METHODS: A total of 48 healthy C57BL/6 adult mice were randomly divided into SCI (SCI model was induced by Allen’s method) and sham 1 groups (n=24 per group). Microarray assay was used to find the differentially expressed miRNAs in the lesion tissues after SCI. An additional 72 mice were randomly divided into sham 2, antagomiR and control groups (n=24 per group). The SCI mice in the later two groups were given the intrathecal injection of antagomir-21a and antagomir, respectively. The locomotor function recovery of hind limbs was evaluated by Basso Mouse Scale score; the expression levels of SCI-related markers in the lesion tissues were detected.
RESULTS AND CONCLUSION: The expression level of miR-21a-5p was significantly up-regulated in the lesion tissues after SCI (P < 0.05). AntagomiR-21a inhibited the expression of miR-21a-5p post-SCI. The Basso Mouse Scale scores showed a significant difference between sham and SCI groups (P < 0.05), the scores in the antagomiR-21a group was higher than those in the control group at 7 days postoperatively, and showed a significant difference on day 4 (P < 0.05). Compared with the control group, antagomiR-21a significantly suppressed the expression levels of fibronectin and CSPGs (P < 0.05), and significantly up-regulated the expression levels of neurotrophic factors (P < 0.05). Our results suggest that there is a up-regulation in the expression level of miR-21a-5p after SCI, and miR-21a-5p knockdown can suppress scar formation and promote secretion of neurotrophic factors, thereby improving motor functional recovery.

中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

Key words: MicroRNAs, Spinal Cord Injuries, Neurturin, Tissue Engineering

中图分类号: