中国神经再生研究(英文版) ›› 2014, Vol. 9 ›› Issue (12): 1186-1194.doi: 10.4103/1673-5374.135325

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

坐骨神经损伤大鼠Wallerian溃变过程中近远端组织的差异基因表达

  

  • 收稿日期:2014-04-23 出版日期:2014-06-24 发布日期:2014-06-24
  • 基金资助:

    国家自然科学基金 (81370982,31170946,81130080) 

Differential gene expression in proximal and distal nerve segments of rats with sciatic nerve injury during Wallerian degeneration

Nan Jiang 1, 2, Huaiqin Li 1, Yi Sun 2, Dexin Yin 2, Qin Zhao 3, Shusen Cui 2, Dengbing Yao 1   

  1. 1 Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Nantong, Jiangsu Province, China
    2 China-Japan Union Hospital of Jilin University, Changchun, Jilin Province, China
    3 Key Laboratory of People’s Liberation Army, Institute of Orthopedics, Chinese PLA General Hospital, Beijing, China
  • Received:2014-04-23 Online:2014-06-24 Published:2014-06-24
  • Contact: Shusen Cui, M.D., Ph.D., China-Japan Union Hospital of Jilin University, Changchun 130033, Jilin Province, China, sscui916@126.com. Dengbing Yao, M.D., Ph.D., Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Nantong 226001, Jiangsu Province, China, yaodb@ntu.edu.cn.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 81370982, 31170946; Key Program, Grant No. 81130080; the Priority Academic Program Development of Jiangsu Higher Education Institutions in China.

摘要:

在神经损伤和修复过程中,Wallerian溃变为神经再生创造了有利的条件,目前尽管对大鼠坐骨神经损伤后Wallerian溃变过程中远端神经组织的基因表达已有了深入的报道,但相关基因的作用机制尚不明确。为此,实验设计了运用分子生物学技术与生物信息学技术,全面系统地分析大鼠坐骨神经损伤后0,4,7,14,21,28 d,Wallerian溃变过程中近远端神经组织的差异基因表达及差异基因的作用机制。基因芯片分析结果显示,大鼠坐骨神经损伤后主要在7-14 d近远端神经组织中有6,000多个差异基因表达,主要包括神经元分化、神经发育和再生、细胞因子合成、轴突再生等相关基因,涉及细胞应激反应、细胞凋亡、炎症反应、免疫反应、信号转导、细胞增殖等个生物学过程。通过实时定量PCR、蛋白免疫印迹和免疫组织化学的方法,分别从基因、蛋白和细胞水平,鉴定了其中的4个基因:水通道蛋白4、白细胞介素1受体蛋白1、基质金属蛋白酶12和编码许旺细胞的成髓鞘蛋白PRX共4个基因,结果与上述基因芯片分析结果一致。实验成功检测并验证了坐骨神经损伤Wallerian溃变过程中近远端组织的差异基因表达,并分析了这些差异基因的在Wallerian溃变过程中近端和远端组织间的动态生物学过程,为深入研究神经损伤及修复过程中WD提供一个可靠的平台。

关键词: 神经再生, 周围神经损伤, 沃勒溃变, 坐骨神经损伤, 基因芯片, 表达谱, 生物学过程, 大鼠, 国家自然科学基金

Abstract:

Wallerian degeneration is a subject of major interest in neuroscience. A large number of genes are differentially regulated during the distinct stages of Wallerian degeneration: transcription factor activation, immune response, myelin cell differentiation and dedifferentiation. Although gene expression responses in the distal segment of the sciatic nerve after peripheral nerve injury are known, differences in gene expression between the proximal and distal segments remain unclear. In the present study in rats, we used microarrays to analyze changes in gene expression, biological processes and signaling pathways in the proximal and distal segments of sciatic nerves undergoing Wallerian degeneration. More than 6,000 genes were differentially expressed and 20 types of expression tendencies were identified, mainly between proximal and distal segments at 7–14 days after injury. The differentially expressed genes were those involved in cell differentiation, cytokinesis, neuron differentiation, nerve development and axon regeneration. Furthermore, 11 biological processes were represented, related to responses to stimuli, cell apoptosis, inflammatory response, immune response, signal transduction, protein kinase activity, and cell proliferation. Using real-time quantitative PCR, western blot analysis and immunohistochemistry, microarray data were verified for four genes: aquaporin-4, interleukin 1 receptor-like 1, matrix metalloproteinase-12 and periaxin. Our study identifies differential gene expression in the proximal and distal segments of a nerve during Wallerian degeneration, analyzes dynamic biological changes of these genes, and provides a useful platform for the detailed study of nerve injury and repair during Wallerian degeneration.

Key words: nerve regeneration, peripheral nerve injury, Wallerian degeneration, sciatic nerve injury, microarray, expression profiling, biological process, rat, NSFC grant, neural regeneration