發(fā)布時間：2019-01-02 18:46

【摘要】：目的：通過構(gòu)建大鼠周圍神經(jīng)損傷模型,比較感覺和運動神經(jīng)束Wallerian變性前后差異表達蛋白,篩選感覺和運動神經(jīng)束特異表達蛋白為周圍神經(jīng)趨化性再生機制的研究提供依據(jù)。方法：選擇成年雄性8周齡Wistar大鼠60只,分成3組,其中A、B兩組分別行股神經(jīng)橫斷模型(n=20)和神經(jīng)根損傷模型(n=20)的建立,C組作為神經(jīng)根損傷模型的正常對照組(n=20),股神經(jīng)橫斷模型選擇對側(cè)正常神經(jīng)作為對照。7d后各組分別進行取材,并應用iTRAQ(isobaric tags for relative and absolute quantitation,同位素標記相對和絕對定量技術)聯(lián)合LC-MS/MS (liquid chromatography-tandem mass spectrometry,液相色譜-串聯(lián)質(zhì)譜)對各組樣本提取的蛋白進行同位素標記、質(zhì)譜分析及差異蛋白的鑒定,應用DAVID (DAVID Bioinformatics Resources)對所鑒定出的差異蛋白進行GO(Gene ontology)功能和KEGG Pathway通路的富集,比較Wallerian變性后感覺和運動神經(jīng)的差異表達蛋白,篩選與周圍神經(jīng)趨化性再生機制相關的蛋白。結(jié)果：(1)在股神經(jīng)損傷模型中,共鑒定出3358個差異表達蛋白,其中股神經(jīng)肌支Wallerian變性前后共鑒定出293個差異蛋白,隱神經(jīng)變性前后共鑒定出417個差異蛋白,通過交集分析,肌支變性前后特異的差異蛋白有107個,隱神經(jīng)變性前后特異的差異蛋白有231個。(2)在神經(jīng)根損傷模型中,共鑒定出4312個差異表達蛋白,腹側(cè)神經(jīng)根變性前后共鑒定出637個差異蛋白,背側(cè)神經(jīng)根變性前后共鑒定出626個差異蛋白,通過交集分析,腹側(cè)神經(jīng)根變性前后特異的差異蛋白有195個,背側(cè)神經(jīng)根變性前后特異的差異蛋白有184個。結(jié)論：應用iTRAQ技術檢測大鼠感覺和運動神經(jīng)纖維變性前后表達差異蛋白,篩選感覺和運動神經(jīng)變性后特異表達的蛋白,為周圍神經(jīng)趨化性再生的機制研究提供了依據(jù)。
[Abstract]:Aim: to establish a rat model of peripheral nerve injury, compare the differentially expressed proteins between sensory and motor tract Wallerian before and after denaturation, and screen the specific expression proteins of sensory and motor nerve bundles to provide evidence for the study of the mechanism of peripheral nerve chemotaxis regeneration. Methods: sixty adult male 8-week-old Wistar rats were randomly divided into 3 groups. The femoral nerve transection model (nong20) and the nerve root injury model (nnt20) were established in group A and B, respectively. Group C was used as the normal control group of nerve root injury model (NN20), and the contralateral normal nerve was selected as control in the model of femoral nerve transection. After 7 days, the samples were taken from each group and iTRAQ (isobaric tags for relative and absolute quantitation, was used. The relative and absolute quantitative techniques of isotopic labeling combined with LC-MS/MS (liquid chromatography-tandem mass spectrometry, liquid Chromatography-tandem Mass Spectrometry) were used to label the proteins extracted from each group of samples by isotope labeling, mass spectrometry analysis and differential protein identification. DAVID (DAVID Bioinformatics Resources) was used to enrich the GO (Gene ontology) function and KEGG Pathway pathway of the identified differentially expressed proteins, to compare the differentially expressed proteins of sensory and motor nerves after Wallerian degeneration, and to screen the proteins related to the mechanism of chemotaxis regeneration of peripheral nerve. Results: (1) in the model of femoral nerve injury, 3358 differentially expressed proteins were identified, including 293 differentially expressed proteins before and after denaturation of femoral nerve muscle branch and 417 differential proteins before and after degeneration of saphenous nerve. There were 107 differentially expressed proteins before and after muscular branch degeneration and 231 differential proteins before and after saphenous nerve degeneration. (2) 4312 differentially expressed proteins were identified in the model of nerve root injury. 637 differential proteins were identified before and after ventral nerve root degeneration, and 626 differential proteins were identified before and after dorsal nerve root degeneration. There were 184 specific differential proteins before and after dorsal nerve root degeneration. Conclusion: using iTRAQ technique to detect the differential protein expression before and after sensory and motor nerve fiber degeneration in rats, and to screen the specific protein expression after sensory and motor nerve degeneration, which provides the basis for the study of the mechanism of peripheral nerve chemotaxis regeneration.
【學位授予單位】：中國人民解放軍醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R651.3

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本文編號：2398848