本文選題：創(chuàng)傷性腦損傷　切入點(diǎn)：NT-3殼聚糖支架　出處：《首都醫(yī)科大學(xué)》2014年碩士論文

【摘要】：研究背景：創(chuàng)傷性腦損傷后，，導(dǎo)致神經(jīng)元死亡或丟失，造成多種神經(jīng)功能缺陷，其中海馬損傷導(dǎo)致認(rèn)知功能缺陷嚴(yán)重影響了患者的空間學(xué)習(xí)與記憶功能。傳統(tǒng)的觀點(diǎn)認(rèn)為中樞神經(jīng)通路是固定的、不能改變的。中樞神經(jīng)死亡后不能再生，由膠質(zhì)細(xì)胞替代丟失的神經(jīng)細(xì)胞。近年研究表明在缺血損傷等條件下，成年大鼠腦海馬內(nèi)存在的內(nèi)源性神經(jīng)前體細(xì)胞可以不斷增殖產(chǎn)生新生神經(jīng)元。這為修復(fù)中樞神經(jīng)損傷提供了理論基礎(chǔ)。 研究目的：創(chuàng)傷性腦損傷后移植入NT-3殼聚糖支架材料，研究損傷腦組織再生及損傷區(qū)邊緣瘢痕情況；觀察損傷區(qū)神經(jīng)再生過(guò)程以及損傷區(qū)內(nèi)新生神經(jīng)元突觸形成，并參與腦神經(jīng)環(huán)路重建的研究。 實(shí)驗(yàn)方法：本研究采用生物吸除裝置造成大鼠腦海馬CA1及以上皮層機(jī)械性損傷做為創(chuàng)傷性腦損傷的實(shí)驗(yàn)?zāi)Ｐ汀?shí)驗(yàn)動(dòng)物隨機(jī)分為三組：分別是單純損傷組、單純殼聚糖支架組、NT-3殼聚糖支架組。分別于術(shù)后3d、7d、14d、28d和60d，各實(shí)驗(yàn)組取4只動(dòng)物灌殺取腦，冰凍切片免疫化學(xué)組織染色觀察。NF染色觀察損傷區(qū)內(nèi)再生的神經(jīng)元及神經(jīng)纖維的情況；GFAP染色觀察損傷邊緣膠質(zhì)瘢痕情況。Nestin免疫組織化學(xué)染色觀察損傷區(qū)內(nèi)神經(jīng)前體細(xì)胞增殖情況；βtubulinⅢ免疫組織化學(xué)染色觀察損傷區(qū)內(nèi)分化的不成熟神經(jīng)元；MAP2免疫組織化學(xué)染色觀察損傷區(qū)內(nèi)分化成熟的神經(jīng)元。術(shù)后30d、60d灌殺動(dòng)物，利用免疫電鏡技術(shù)和免疫組織化學(xué)染色觀察損傷區(qū)內(nèi)再生神經(jīng)元的突觸超微結(jié)構(gòu)。術(shù)后60d，應(yīng)用神經(jīng)示蹤方法將BDA-FITC注射到損傷對(duì)側(cè)CA3區(qū)，熒光顯微鏡下觀察示蹤劑走形情況。 研究結(jié)果：免疫組織化學(xué)結(jié)果顯示：創(chuàng)傷性腦損傷后60天，各組損傷區(qū)內(nèi)都可見不同數(shù)量神經(jīng)再生，統(tǒng)計(jì)數(shù)值后發(fā)現(xiàn)：NT-3殼聚糖支架組同單純殼聚糖組、單純損傷組存在明顯的統(tǒng)計(jì)學(xué)差異；同時(shí)觀察損傷邊緣的膠質(zhì)瘢痕情況發(fā)現(xiàn)NT-3殼聚糖支架組GFAP陽(yáng)性細(xì)胞數(shù)明顯少于單純損傷組和單純殼聚糖支架組。以上結(jié)果說(shuō)明創(chuàng)傷性腦損傷后，移植NT-3殼聚糖支架可以促進(jìn)損傷區(qū)內(nèi)神經(jīng)再生，抑制損傷區(qū)邊緣膠質(zhì)瘢痕的形成。進(jìn)一步研究移植NT-3殼聚糖支架到損傷區(qū)后，觀察神經(jīng)前體細(xì)胞增殖分化為成熟神經(jīng)元的過(guò)程。研究發(fā)現(xiàn)移植NT-3殼聚糖支架組，術(shù)后3天時(shí)nestin陽(yáng)性細(xì)胞被激活，7天時(shí)nestin陽(yáng)性細(xì)胞數(shù)達(dá)高峰，到14天出現(xiàn)下降趨勢(shì)。觀察發(fā)現(xiàn)腦損傷后14天βtubulinⅢ陽(yáng)性細(xì)胞數(shù)增多，28天后陽(yáng)性細(xì)胞數(shù)下降。研究發(fā)現(xiàn)腦損傷后28天MAP2陽(yáng)性細(xì)胞數(shù)逐漸增多，此增長(zhǎng)趨勢(shì)延續(xù)到腦損傷后60天。統(tǒng)計(jì)數(shù)值發(fā)現(xiàn)：NT-3殼聚糖支架組nestin、βtubulinⅢ和MAP2陽(yáng)性細(xì)胞數(shù)同其他對(duì)照組存在明顯的統(tǒng)計(jì)學(xué)差異。移植NT-3殼聚糖支架組損傷后30天，通過(guò)免疫電鏡方法觀察損傷區(qū)內(nèi)出現(xiàn)BrdU+MAP2+雙陽(yáng)性的新生神經(jīng)元，并形成成熟突觸結(jié)構(gòu)。熒光顯微鏡下觀察到BDA+神經(jīng)細(xì)胞通過(guò)右側(cè)海馬CA3錐體細(xì)胞的軸突沿著海馬聯(lián)合到達(dá)左側(cè)CA1區(qū)，在損傷區(qū)內(nèi)觀察到BDA+細(xì)胞，并且部分BDA+細(xì)胞表達(dá)MAP2+。 研究結(jié)論：NT-3殼聚糖支架可以促進(jìn)損傷區(qū)內(nèi)神經(jīng)再生，抑制損傷周邊膠質(zhì)瘢痕的形成。損傷區(qū)內(nèi)神經(jīng)前體細(xì)胞能夠增殖分化為成熟神經(jīng)元并形成神經(jīng)突觸，參與腦神經(jīng)網(wǎng)絡(luò)的修復(fù)重建。
[Abstract]:Background : After traumatic brain injury , the death or loss of neurons leads to a variety of neurological deficits , in which the damage of the hippocampus leads to a severe impairment of cognitive function . The traditional view suggests that the central nervous pathway is fixed and cannot be changed . In recent years , the endogenous neural progenitor cells present in the hippocampus of adult rats can proliferate and produce new neurons . This is a theoretical basis for repairing the central nervous system injury .

Objective : To study the regeneration of injured brain tissue and the scar of injured area after traumatic brain injury .
Observe the nerve regeneration process in the injured area and the synaptic formation of neonatal neurons in the injured area , and participate in the study of the reconstruction of the brain nerve loop .

The experimental animal model of traumatic brain injury was studied by means of biosorption device . The experimental animals were randomly divided into three groups : simple injury group , chitosan stent group and NT - 3 chitosan stent group . The experimental animals were divided into three groups : simple injury group , simple chitosan stent group and NT - 3 chitosan stent group .
GFAP staining was used to observe the glial scar in the injured area . Nestin immunohistochemistry staining was used to observe the proliferation of neural progenitor cells in the injured area .
Non - mature neurons in the injured area were observed by immunohistochemical staining of 尾 - 鈪⑩參 .
The synaptic ultrastructure of regenerated neurons in the injured area was observed by immunohistochemical staining of MAP2 . After the operation , the synaptic ultrastructure of the regenerated neurons in the injured area was observed by immunoelectron microscopy and immunohistochemical staining . After operation 60 days after operation , the experimental tracer was injected into the injured contralateral CA area under the fluorescence microscope .

The results showed that after traumatic brain injury 60 days after traumatic brain injury , different numbers of nerve regeneration were observed in each group , and it was found that NT - 3 chitosan stent group had obvious statistical difference compared with simple chitosan group and simple injury group .
It was found that the number of GFAP - positive cells in NT - 3 chitosan stent group was significantly lower than that of simple injury group and simple chitosan stent group .

Conclusion : NT - 3 chitosan scaffold can promote nerve regeneration in the injured area and inhibit the formation of glial scar in the injured peripheral area . The neural progenitor cells in the injured area can proliferate into mature neurons and form neuroses , and participate in the repair and reconstruction of cranial nerve network .

【學(xué)位授予單位】：首都醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R651.15

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

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2 張小年;張皓;朱鏞連;;創(chuàng)傷性顱腦損傷的流行病學(xué)概況[J];中國(guó)康復(fù)理論與實(shí)踐;2005年12期

本文編號(hào)：1708660