【摘要】：目的：通過(guò)構(gòu)建大鼠周?chē)窠?jīng)損傷模型,比較感覺(jué)和運(yùn)動(dòng)神經(jīng)束Wallerian變性前后差異表達(dá)蛋白,篩選感覺(jué)和運(yùn)動(dòng)神經(jīng)束特異表達(dá)蛋白為周?chē)窠?jīng)趨化性再生機(jī)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(duì)照組(n=20),股神經(jīng)橫斷模型選擇對(duì)側(cè)正常神經(jīng)作為對(duì)照。7d后各組分別進(jìn)行取材,并應(yīng)用iTRAQ(isobaric tags for relative and absolute quantitation,同位素標(biāo)記相對(duì)和絕對(duì)定量技術(shù))聯(lián)合LC-MS/MS (liquid chromatography-tandem mass spectrometry,液相色譜-串聯(lián)質(zhì)譜)對(duì)各組樣本提取的蛋白進(jìn)行同位素標(biāo)記、質(zhì)譜分析及差異蛋白的鑒定,應(yīng)用DAVID (DAVID Bioinformatics Resources)對(duì)所鑒定出的差異蛋白進(jìn)行GO(Gene ontology)功能和KEGG Pathway通路的富集,比較Wallerian變性后感覺(jué)和運(yùn)動(dòng)神經(jīng)的差異表達(dá)蛋白,篩選與周?chē)窠?jīng)趨化性再生機(jī)制相關(guān)的蛋白。結(jié)果：(1)在股神經(jīng)損傷模型中,共鑒定出3358個(gè)差異表達(dá)蛋白,其中股神經(jīng)肌支Wallerian變性前后共鑒定出293個(gè)差異蛋白,隱神經(jīng)變性前后共鑒定出417個(gè)差異蛋白,通過(guò)交集分析,肌支變性前后特異的差異蛋白有107個(gè),隱神經(jīng)變性前后特異的差異蛋白有231個(gè)。(2)在神經(jīng)根損傷模型中,共鑒定出4312個(gè)差異表達(dá)蛋白,腹側(cè)神經(jīng)根變性前后共鑒定出637個(gè)差異蛋白,背側(cè)神經(jīng)根變性前后共鑒定出626個(gè)差異蛋白,通過(guò)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(gè),背側(cè)神經(jīng)根變性前后特異的差異蛋白有184個(gè)。結(jié)論：應(yīng)用iTRAQ技術(shù)檢測(cè)大鼠感覺(jué)和運(yùn)動(dòng)神經(jīng)纖維變性前后表達(dá)差異蛋白,篩選感覺(jué)和運(yùn)動(dòng)神經(jīng)變性后特異表達(dá)的蛋白,為周?chē)窠?jīng)趨化性再生的機(jī)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(xué)位授予單位】：中國(guó)人民解放軍醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R651.3

【參考文獻(xiàn)】

相關(guān)期刊論文 前3條

1 曾嶸,夏其昌;蛋白質(zhì)組學(xué)研究進(jìn)展與趨勢(shì)[J];中國(guó)科學(xué)院院刊;2002年03期

2 王林纖;戴勇;涂植光;;iTRAQ標(biāo)記技術(shù)與差異蛋白質(zhì)組學(xué)的生物標(biāo)志物研究[J];生命的化學(xué);2010年01期

3 褚庭綱;高偉陽(yáng);黃金龍;李志杰;陳星隆;李曉陽(yáng);;新生SD大鼠坐骨神經(jīng)Wallerian變性規(guī)律及其S-100蛋白表達(dá)和雪旺細(xì)胞增殖研究[J];溫州醫(yī)學(xué)院學(xué)報(bào);2007年06期

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