中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2018, Vol. 13 ›› Issue (12): 2182-2190.doi: 10.4103/1673-5374.241469

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

不同鼠龄段SD大鼠坐骨神经转录组测序分析

  

  • 收稿日期:2018-08-20 出版日期:2018-12-15 发布日期:2018-12-15
  • 基金资助:

    国家自然科学基金资助（81201546）(YXL)；广东省自然科学基金博士启动项目（2017A030310302）(ZWZ)；广东省医学科研基金（A2016018）(BH)；中国广东省科学技术项目（2016A010103012）(JHL)

Analysis of transcriptome sequencing of sciatic nerves in Sprague-Dawley rats of different ages

Jiang-Hui Liu1, Qing Tang2, Xiang-Xia Liu2, Jian Qi3, Rui-Xi Zeng2, Zhao-Wei Zhu2, Bo He4, Yang-Bin Xu2   

  1. 1 Department of Emergency, First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong Province, China
    2 Department of Plastic Surgery, First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong Province, China
    3 Department of Orthopedics and Microsurgery, First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong Province, China
    4 Department of Orthopedics, Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong Province, China
  • Received:2018-08-20 Online:2018-12-15 Published:2018-12-15
  • Contact: Zhao-Wei Zhu, MD, PhD, nmtmemoir@aliyun.com; Bo He, MD, PhD, hebodoc@gmail.com.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 81201546 (to YXL); the Doctoral Start-up Program of Natural Science Foundation of Guangdong Province of China, No. 2017A030310302 (to ZWZ); the Medical Scientific Research Foundation of Guangdong Province of China, No. A2016018 (to BH); the Science and Technology Project of Guangdong Province of China, No.2016A010103012 (to JHL).

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摘要：

衰老引起许旺细胞活性的下降可影响哺乳动物周围神经损伤后的再生，因此有必要明确与鼠龄有关的影响周围神经生物学功能的基因表达情况。将健康雄性SD大鼠按鼠龄分为幼年组(1周龄)和成年组(12月龄)，每组10只。实验取每组4只动物的双侧坐骨神经，用第2代高通量全RNA测序(Next Generation Sequencing，NGS)和生物信息学的方法分析两组坐骨神经组织mRNA表达情况，筛选差异表达mRNAs(Differentially Expressed mRNA，DEmRNAs)；每组取6只大鼠，根据生物信息学分析的结果随机选取18组DEmRNAs进行qRT-PCR检测；结果显示：(1)与1周龄大鼠相比，12月龄大鼠有3608组DEmRNAs，其中2684组上调，924组下调，其功能主要涉及细胞活性、增殖、分化、再生和髓鞘形成等；(2)上调最明显的基因是Thrsp(Log2FC＝9.01，P＜0.05)，下调最明显的基因是COL2A1(Log2FC＝-8 89，P < 0.05) ；(3) Go分析表明，DEmRNAs主要集中在寡糖结合、核苷酸结合寡聚结构域、信号转导通路、肽转运ATP酶活性等生物学过程；(4) KEGG分析显示，大鼠随着鼠龄的增长，DEmRNAs主要富集于甾体生物合成、金黄色葡萄球菌感染及移植物抗宿主病等通路；(5)采用Spearman相关系数法评价NGS的准确性发现，NGS结果与QRT-PCR结果的变化趋势呈正相关(R2=0.5477，P ＜ 0.05)；(6)上述数据可证实，成年 (12月龄)与幼年(1周龄)大鼠坐骨神经的基因表达存在差异，周围神经细胞和微环境随鼠龄的增长而变化，进而可能影响到周围神经的功能和损伤后的修复效果。

orcid:0000-0003-4469-5417(Zhao-Wei Zhu)
        0000-0002-9690-4527(Bo He)

关键词: 周围神经损伤, 老年, SD大鼠, 转录组, 测序, mRNA, 鼠龄, 许旺细胞, 神经再生

Abstract:

An aging-induced decrease in Schwann cell viability can affect regeneration following peripheral nerve injury in mammals. It is therefore necessary to investigate possible age-related changes in gene expression that may affect the biological function of peripheral nerves. Ten 1-week-old and ten 12-month-old healthy male Sprague-Dawley rats were divided into young (1 week old) and adult (12 months old) groups according to their ages. mRNA expression in the sciatic nerve was compared between young and adult rats using next-generation sequencing (NGS) and bioinformatics (n = 4/group). The 18 groups of differentially expressed mRNA (DEmRNAs) were also tested by quantitative reverse transcription polymerase chain reaction (n = 6/group). Results revealed that (1) compared with young rats, adult rats had 3608 groups of DEmRNAs. Of these, 2684 were groups of upregulated genes, and 924 were groups of downregulated genes. Their functions mainly involved cell viability, proliferation, differentiation, regeneration, and myelination. (2) The gene with the most obvious increase of all DEmRNAs in adult rats was Thrsp (log2FC = 9.01, P < 0.05), and the gene with the most obvious reduction was Col2a1 (log2FC = –8.89, P < 0.05). (3) Gene Ontology analysis showed that DEmRNAs were mainly concentrated in oligosaccharide binding, nucleotide-binding oligomerization domain containing one signaling pathway, and peptide-transporting ATPase activity. (4) Analysis using the Kyoto Encyclopedia of Genes and Genomes showed that, with increased age, DEmRNAs were mainly enriched in steroid biosynthesis, Staphylococcus aureus infection, and graft-versus-host disease. (5) Spearman’s correlation coefficient method for evaluating NGS accuracy showed that the NGS results and quantitative reverse transcription polymerase chain reaction results were positively correlated (rs = 0.74, P < 0.05). These findings confirm a difference in sciatic nerve gene expression between adult and young rats, suggesting that, in peripheral nerves, cells and the microenvironment change with age, thus influencing the function and repair of peripheral nerves.

Key words: peripheral nerve injury, aging, Sprague-Dawley rat, transcriptome, sequencing, mRNA, rat age, Schwann cells, neural regeneration