大鼠脊神经前、后根损伤后的蛋白表达差异

发布时间：2018-04-12 09:25

本文选题：瓦勒氏变性 + iTRAQ　；参考：《中国人民解放军医学院》2017年硕士论文

【摘要】：目的:周围神经损伤的发生率高。神经损伤后会发生瓦勒氏变性即神经损伤远端轴突及髓鞘发生崩解,继而炎细胞迁移、增殖、浸润等一系列过程。瓦勒氏变性会严重影响神经的功能,但同时对神经的再生也起到了关键的作用。周围神经损伤后再生过程中常会出现神经错接现象,即运动神经纤维长入感觉神经纤维支配的靶器官的现象,这可能与周围神经运动神经纤维与感觉神经纤维蛋白存在差异有关。但是相关研究表明,运动神经和感觉神经瓦勒氏变性过程中蛋白质的差异对周围神经趋化性再生有更加重要的作用。因此,本研究分析脊神经前、后根损伤7天前后的差异蛋白,为周围神经趋化性再生提供实验基础。方法:成年雌性SD大鼠60只,质量220～250g。将右侧脊神经前、后根横断,左侧作为对照组不作处理。造模7天后,取离断的脊神经前根远断端为运动神经瓦勒氏变性组,取离断的脊神经后根近断端为感觉神经瓦勒氏变性组,取健侧脊神经前根为正常运动神经对照组,取健侧的脊神经后根为正常感觉神经对照组,提取蛋白,为保证实验可重复性,每组样本分为两批,共计8个样本。以上样本应用 iTRAQ (isobaric tags for relative and absolute quantitation,同位素标记相对和绝对定量)技术鉴定差异蛋白,并应用GO(Geneontology,基因本体论)和KEGG(Kyoto Encyclopedia of Genes and Genomes,京都基因与基因组百科全书)对差异蛋白进行分析。结果:应用iTRAQ共鉴定出4662个蛋白。根据两次交集结果,以至少包含两个独特肽段并且p0.05为标准筛选差异蛋白。与正常的脊神经前根相比,损伤7天后的脊神经前根共鉴定出17个差异蛋白,其中有11个表达升高,6个表达降低。与正常的脊神经后根相比,损伤7天后的脊神经后根共鉴定出2个差异蛋白,均为下调蛋白。通过交集分析发现,脊神经前根与脊神经后根损伤7天后的差异蛋白无交集,均为特异差异蛋白。结论:本研究筛选出了大鼠脊神经前、后根损伤7天后的差异蛋白,为后续周围神经再生研究提供了数据基础。
[Abstract]:Objective: the incidence of peripheral nerve injury is high.After nerve injury, Vall's degeneration will occur, that is, the distal axon and myelin sheath of nerve injury will disintegrate, and then the inflammatory cells will migrate, proliferate and infiltrate a series of processes.Vall's degeneration can seriously affect nerve function, but also plays a key role in nerve regeneration.In the process of regeneration after peripheral nerve injury, nerve dislocation often occurs, that is, motor nerve fibers grow into the target organs innervated by sensory nerve fibers.This may be related to the difference between peripheral nerve motor nerve fibers and sensory nerve fibrils.But related studies have shown that the difference of protein in the process of Vall's degeneration of motor and sensory nerves plays a more important role in the regeneration of peripheral nerve chemotaxis.Therefore, we analyzed the differential proteins before and after 7 days of spinal nerve injury to provide experimental basis for peripheral nerve chemotaxis regeneration.Methods: 60 adult female SD rats, 250 g.The anterior and posterior roots of the right spinal nerve were transected, and the left side was not treated as the control group.After 7 days of modeling, the distal end of the disconnected anterior root of the spinal nerve was taken as the motor nerve Vall's degeneration group, the proximal end of the disconnected posterior root of the spinal nerve was taken as the sensory nerve Vall's degeneration group, and the contralateral anterior root of the spinal nerve was taken as the control group of the normal motor nerve.The contralateral posterior root of the spinal nerve was taken as the control group of normal sensory nerve, and protein was extracted. In order to ensure the repeatability of the experiment, each group was divided into two groups, 8 samples in total.The differentially expressed proteins were identified by iTRAQ isobaric tags for relative and absolute quantitative (relative and absolute quantitative) techniques, and the differential proteins were analyzed by KEGG(Kyoto Encyclopedia of Genes and genetics (Genontology) and KEGG(Kyoto Encyclopedia of Genes and Genome (Kyoto Encyclopedia and Genome Encyclopedia).Results: 4662 proteins were identified by iTRAQ.According to the results of two intersections, differential proteins were screened with at least two idiopeptide segments and p0.05 as the standard.Compared with the normal anterior root of spinal nerve, 17 differentially expressed proteins were identified 7 days after injury, of which 11 expressed higher and 6 decreased.Compared with the normal posterior root of spinal nerve, two differentially expressed proteins were identified in the posterior root of spinal nerve 7 days after injury, all of which were down-regulated proteins.It was found that the differential proteins of the anterior root of spinal nerve and the posterior root of spinal nerve were not intersected 7 days after injury, and they were all specific differential proteins.Conclusion: in this study, the differential proteins were screened out 7 days after the injury of the anterior and posterior roots of the spinal nerve in rats, which provided the data basis for the follow-up study of peripheral nerve regeneration.
【学位授予单位】：中国人民解放军医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R651.3

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