發(fā)布時間：2018-04-12 09:25

  本文選題：瓦勒氏變性 + iTRAQ��； 參考：《中國人民解放軍醫(yī)學(xué)院》2017年碩士論文

【摘要】：目的:周圍神經(jīng)損傷的發(fā)生率高。神經(jīng)損傷后會發(fā)生瓦勒氏變性即神經(jīng)損傷遠端軸突及髓鞘發(fā)生崩解,繼而炎細(xì)胞遷移、增殖、浸潤等一系列過程。瓦勒氏變性會嚴(yán)重影響神經(jīng)的功能,但同時對神經(jīng)的再生也起到了關(guān)鍵的作用。周圍神經(jīng)損傷后再生過程中常會出現(xiàn)神經(jīng)錯接現(xiàn)象,即運動神經(jīng)纖維長入感覺神經(jīng)纖維支配的靶器官的現(xiàn)象,這可能與周圍神經(jīng)運動神經(jīng)纖維與感覺神經(jīng)纖維蛋白存在差異有關(guān)。但是相關(guān)研究表明,運動神經(jīng)和感覺神經(jīng)瓦勒氏變性過程中蛋白質(zhì)的差異對周圍神經(jīng)趨化性再生有更加重要的作用。因此,本研究分析脊神經(jīng)前、后根損傷7天前后的差異蛋白,為周圍神經(jīng)趨化性再生提供實驗基礎(chǔ)。方法:成年雌性SD大鼠60只,質(zhì)量220～250g。將右側(cè)脊神經(jīng)前、后根橫斷,左側(cè)作為對照組不作處理。造模7天后,取離斷的脊神經(jīng)前根遠斷端為運動神經(jīng)瓦勒氏變性組,取離斷的脊神經(jīng)后根近斷端為感覺神經(jīng)瓦勒氏變性組,取健側(cè)脊神經(jīng)前根為正常運動神經(jīng)對照組,取健側(cè)的脊神經(jīng)后根為正常感覺神經(jīng)對照組,提取蛋白,為保證實驗可重復(fù)性,每組樣本分為兩批,共計8個樣本。以上樣本應(yīng)用 iTRAQ (isobaric tags for relative and absolute quantitation,同位素標(biāo)記相對和絕對定量)技術(shù)鑒定差異蛋白,并應(yīng)用GO(Geneontology,基因本體論)和KEGG(Kyoto Encyclopedia of Genes and Genomes,京都基因與基因組百科全書)對差異蛋白進行分析。結(jié)果:應(yīng)用iTRAQ共鑒定出4662個蛋白。根據(jù)兩次交集結(jié)果,以至少包含兩個獨特肽段并且p0.05為標(biāo)準(zhǔn)篩選差異蛋白。與正常的脊神經(jīng)前根相比,損傷7天后的脊神經(jīng)前根共鑒定出17個差異蛋白,其中有11個表達升高,6個表達降低。與正常的脊神經(jīng)后根相比,損傷7天后的脊神經(jīng)后根共鑒定出2個差異蛋白,均為下調(diào)蛋白。通過交集分析發(fā)現(xiàn),脊神經(jīng)前根與脊神經(jīng)后根損傷7天后的差異蛋白無交集,均為特異差異蛋白。結(jié)論:本研究篩選出了大鼠脊神經(jīng)前、后根損傷7天后的差異蛋白,為后續(xù)周圍神經(jīng)再生研究提供了數(shù)據(jù)基礎(chǔ)。
[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.
【學(xué)位授予單位】：中國人民解放軍醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R651.3

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