比较蛋白质组学在周围神经感觉束和运动束鉴别中作用的实验研究  

Comparative proteomics study of sensory and motor fascicles of peripheral nerve

作　　者：孟宪宇[1] 路来金[2] 陈焕新[3] MENG Xianyu;LU Laijin;CHEN Huanxin(Department of Orthopedics,The First Affiliated Hospital,Heilongjiang University of Chinese Medicine,Harbin 150040,China)

机构地区：[1]黑龙江中医药大学附属第一医院骨科,黑龙江哈尔滨150040 [2]吉林大学白求恩第一医院手外科,吉林长春130021 [3]佛罗里达大学神经外科系,Florida Gainesville32611

出　　处：《中风与神经疾病杂志》2022年第4期302-305,共4页Journal of Apoplexy and Nervous Diseases

基　　金：教育部春晖计划项目(Z2010040)。

摘　　要：目的寻找周围神经感觉和运动束之间的特异蛋白,为周围神经功能束的鉴别提供标志蛋白。方法分别对Wistar大鼠股神经运动支和隐神经正常状态、在Sunderland V损伤后8 h和8 d时间点取材,断端远侧各切取5 mm样本,6种样本共18组分离提取蛋白,纯化及定量后应用差异凝胶电泳技术(DIGE)荧光标记蛋白质,凝胶图像扫描,用图像分析软件(DeCyder)比较分析并识别各组间的差异表达蛋白质图谱,选择表达差异大于1.5倍的蛋白点,制备胶上切割、酶切、点靶,MALDI-TOF-Pro质谱仪进行PMF图谱分析鉴定,应用Rt-PCR技术分析验证蛋白质组学研究的结果。结果Annexin V、NF-L、Tec、Serpina3n、PRDX-2、TPM1等6个蛋白点被确切地鉴定。Annexin V在股神经运动支/隐神经的比值是-1.94倍,匹配良好,蛋白稳定出现在所有胶中,P(0.00068)<0.05,具有统计学意义;在Sunderland V损伤后8 h的股神经运动支/隐神经的比值是-2.07倍,匹配良好,蛋白稳定出现在所有胶中,P(0.0053)<0.05,具有统计学意义;在Sunderland V损伤后8 h的股神经运动支/隐神经的比值是-2.20倍,匹配良好,蛋白稳定出现在所有胶中,P(0.0069)<0.05,具有统计学意义。结论周围神经感觉束和运动束之间的比较蛋白质组学表达存在明显差异,Annexin V可作为鉴别周围神经感觉和运动束的高差异标记蛋白。Objective It is important to distinguish between motor and sensory fascicles of the peripheral nerves for nerve alignment in surgery.No biomarkers currently are available for identification of motor or sensory fascicles.The objective of this study is to search the specific proteins between sensory and motor fascicles of peripheral nerves and provide biomarkers for the identification of functional fascicles of peripheral nerves.Methods The normal state of motor branch and saphenous nerve of femoral nerve in Wistar rats,and at 8 hours and 8 days after Sunderland V injury were respectively sampled.Five mm long samples were taken from the distal side of the broken end,and a total of 18 groups of proteins were isolated from 6 samples.After purification and quantification,differential gel electrophoresis(DIGE)was used to label the proteins,gel image was scanned,and image analysis software(DeCyder)was used to compare and identify the differentially expressed proteins in each group.Protein spots with more than 1.5 times of difference in expression were selected to prepare glue-cutting,enzyme-cutting and spot target.PMF chromatogram was analyzed and identified by MALDI-TOF-PRO mass spectrometer,and the results of proteomics were analyzed and compared by RT-PCR.Chi-square tests and t-tests were performed for comparison between motor or sensory nerve groups.Results The data identified 6 proteins that were differentially expressed between motor and sensory fascicles(>1.5-fold,P<0.05),including Annexin V,neurofilament light polypepticle,TEC kinase,serine protease inhibitor A3N,Peroxiredoxin-2,and TPM1.The proteomic results were consistent with the mRNA expression levels of these genes as determined by quantitative reverse transcription polymerase chain reaction.Conclusion There were significant differences in proteomic expression between the peripheral sensory and motor fascicles,and Annexin V can be used as a high-difference marker protein to distinguish the peripheral sensory from motor fascicles.

关 键 词：周围神经 感觉束与运动束 鉴别 蛋白质组学 

分 类 号：R741[医药卫生—神经病学与精神病学]