文章摘要
孟宪宇 ，李秋红 ，刘岩 ，张旭 ，王磊 ，姜巍 ，李凤久 ，路来金 ，陈焕新.周围神经感觉束和运动束在神经损伤修复过程中蛋白变化的比较蛋白质组学研究[J].神经损伤功能重建,2023,(10):574-578
周围神经感觉束和运动束在神经损伤修复过程中蛋白变化的比较蛋白质组学研究
Comparative Proteomics Study on the Changes of Proteins in Sensory and Motor Fasciclesof Peripheral Nerve during the Injury and Repairing Process
  
DOI：
中文关键词: 周围神经  感觉束  运动束  再生  蛋白质组学
英文关键词: peripheral nerves  sensory fascicles  motor fascicles  regeneration  proteomics
基金项目:黑龙江省自然科学 基金（No. LH2019H0 50）； 黑龙江省中医药科 研项目（No. ZHY19- 032）； 黑龙江中医药大学教 育教学研究基金项目 （No. XJJ2014006）； 高等学校博士学科点 专 项 科 研 基 金（No. 20132327120004）； 哈尔滨市科技计划自 筹经费项目（No.ZC2 022ZJ003025）； 教育部高等学校中西 医结合类专业教学指 导委员会 2022 年度 教 育 研 究 课 题（No. 2022-ZC16）； 国家考试中心中医药 考试科研课题 2022 年度（No. 51）
作者单位
孟宪宇1 ，李秋红1 ，刘岩1 ，张旭1 ，王磊1 ，姜巍1 ，李凤久1 ，路来金2 ，陈焕新3 1. 黑龙江中医药大学 附属第一医院骨科 2. 吉林大学白求恩第 一医院手外科 3. 佛罗里达大学神经 外科系 
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中文摘要:
      目的：探寻周围神经感觉和运动束损伤后不同时间点的蛋白变化，为周围神经再生研究提供标志 蛋白。方法：制备SD大鼠股神经运动束和隐神经（感觉束）在正常状态、Sunderland V损伤8 h和8 d时间 点模型；神经断端远侧5 mm样本取材，6种样本共18组；分离、提取、纯化及定量蛋白后，应用差异凝胶电 泳技术荧光标记蛋白质，凝胶图像扫描，选择差异表达＞1.5倍的蛋白点定义为高差异表达蛋白；用图像 分析软件BVA分析显示9块凝胶平均分离1586个蛋白点，胶上切割、酶切、点靶，质谱仪进行肽质量指纹 谱（PMF）分析鉴定，识别各组间的高差异表达蛋白质图谱。应用逆转录PCR（RT-PCR）技术分析验证蛋 白质组学研究的结果。结果：共有12个高差异表达蛋白点被确切鉴定：TPM1、Serpina10、NF-L、DRP-2、 CaBP1、Serpina3n、Peroxiredoxin-2、Gng7、HSP70、LDHB、Enolase2、PDIA3。 其 中 ，Serpina10、TPM1、 NF-L、DRP-2、CaBP1是隐神经正常状态、损伤8 h和8 d时间点时比较蛋白质组学高差异蛋白；Serpina10、 Serpina3n、Peroxiredoxin-2、Gng7、HSP70、LDHB、Enolase2、CaBP1、PDIA3是股神经运动束正常状态、损 伤8 h和8 d时间点时比较蛋白质组学的高差异蛋白。Serpina10 和CaBP1是隐神经和股神经运动束损伤 后多时间点比较蛋白质组学均出现的高差异表达的蛋白。蛋白质组学的结果与RT-PCR检测的mRNA 转录水平相一致。结论：周围神经感觉束、运动束损伤后不同时间的比较蛋白质组学表达存在明显差异， Serpina10 和CaBP1作为高差异表达蛋白，可能为周围神经再生研究提供新的方法。
英文摘要:
      To investigate the changes in proteins in peripheral nerve sensory and motor fascicles at different time points after injury, and to provide markers for peripheral nerve regeneration research. Meth⁃ ods: SD rats were used to prepare normal femoral nerve sensory and saphenous nerve models, as well as 8 h and 8 d post-injury of Sunderland V models. Tissue samples were collected from 5 mm distal ends of broken nerve ends, with 18 groups of samples in total. After separation, extraction, purification, and quantification of proteins, differential gel electrophoresis (DIGE) technology was used to fluorescently label proteins. Gel images were scanned, and high-differential protein spots were defined as those with a fold change greater than 1.5. Image analysis software BVA was used to analyze the gel images and display an average of 1586 protein spots separated on nine gels. Gel cuts, enzyme digestion, and point target were performed, and mass spectrometry was used to analyze the peptide mass fingerprinting (PMF) profile of the identified proteins. The results of image analysis were validated by reverse transcription polymerase chain reaction (RT-PCR) technology. Re⁃ sults: A total of 12 protein spots were accurately identified: TPM1, Serpina10, NF-L, DRP-2, CaBP1, Serpina3n, Peroxiredoxin-2, Gng7, HSP70, LDHB, Enolase2, and PDIA3. Among these, Serpina10, TPM1, NF-L, DRP-2, CaBP1 were identified as high-differential proteins in SD rats with saphenous nerves at both the normal and injured time points; Serpina10, Serpina3n, Peroxiredoxin-2, Gng7, HSP70, LDHB, Enolase2, CaBP1, and PDIA3 were identified as high-differential proteins in SD rats with femoral nerve motor fascicles at both the normal and injured time points. Serpina10 and CaBP1 were identified as high-differential proteins in SD rats with saphenous nerves and femoral nerve motor fascicles at multiple time points after injury. The proteomic results were consistent with the mRNA transcription levels of these genes as determined by Rt-PCR. Conclusion: There are significant differences in protein expression between peripheral nerve sensory and motor fascicles at different time points after injury. Serpina10 and CaBP1, as high-difference marker proteins, were expected to provide new methods for research on peripheral nerve injury and regeneration.
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