本文選題：神經(jīng)干細(xì)胞 + 腦缺血��； 參考：《廈門(mén)大學(xué)》2014年碩士論文

【摘要】：目的：通過(guò)建立腦缺血再灌注大鼠模型,觀察神經(jīng)干細(xì)胞(Neural Stem Cells, NSCs)移植對(duì)腦缺血再灌注大鼠紋狀體梗死區(qū)域新生細(xì)胞以及側(cè)腦室下區(qū)內(nèi)源性NSCs增殖分化的影響。 方法：通過(guò)提取孕14天胎鼠海馬,利用懸浮培養(yǎng)法分離培養(yǎng)鼠源性NSCs。缺血模型構(gòu)建成功后24小時(shí),在腦立體定位儀引導(dǎo)下向左側(cè)紋狀體注射N(xiāo)SCs。將實(shí)驗(yàn)大鼠隨機(jī)分為正常組、假手術(shù)組、缺血模型組(以下簡(jiǎn)稱缺血組)、缺血移植PBS組(以下簡(jiǎn)稱PBS組)和缺血移植神經(jīng)干細(xì)胞組(以下簡(jiǎn)稱NSCs組),分別于第3天、7天、14天進(jìn)行觀察。利用7分制的評(píng)估標(biāo)準(zhǔn)對(duì)缺血引起的神經(jīng)功能缺失狀況進(jìn)行評(píng)估；采用Foot-fault的方法對(duì)所有動(dòng)物損傷的前肢進(jìn)行了檢測(cè)；TTC觀察梗塞面積的變化;尼氏染色結(jié)果觀察缺血區(qū)細(xì)胞形態(tài)和結(jié)構(gòu)；Tunel法檢測(cè)細(xì)胞凋亡并進(jìn)行凋亡陽(yáng)性細(xì)胞計(jì)數(shù)；Brdu/NeuN、Brdu/GFAP熒光雙標(biāo)法觀察紋狀體以及側(cè)腦室下區(qū)(SVZ)雙標(biāo)陽(yáng)性細(xì)胞的表達(dá)情況；統(tǒng)計(jì)相關(guān)區(qū)域陽(yáng)性細(xì)胞數(shù)。 結(jié)果： 1、NSCs組較PBS組的右側(cè)肢體偏癱程度減輕。 2、TTC染色：NSCs組較PBS組的腦梗死面積明顯減小。 3、尼氏染色：PBS組的缺血區(qū)有大量的神經(jīng)元壞死,損傷主要集中在紋狀體,NSCs組神經(jīng)細(xì)胞的變性損傷范圍明顯減少。 4、Tunel(細(xì)胞凋亡染色)：NSCs組在各時(shí)間點(diǎn)的凋亡陽(yáng)性細(xì)胞數(shù)均少于PBS組。 5、免疫熒光雙標(biāo)：SVZ區(qū)內(nèi)源性神經(jīng)干細(xì)胞在第3天開(kāi)始出現(xiàn)增殖,第7天到達(dá)最高峰,第14天明顯減少。其中,NSCs組較PBS組的Brdu/NeuN數(shù)目明顯增多,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。NSCs組較PBS組的Brdu/GFAP數(shù)目明顯減少,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。PBS組紋狀體可見(jiàn)大量Brdu/GFAP陽(yáng)性細(xì)胞,而NSCs組Brdu/GFAP陽(yáng)性細(xì)胞較少。 結(jié)論： 1、移植NSCs后,神經(jīng)功能缺失癥狀以及協(xié)調(diào)能力得到了明顯的改善。 2、移植NSCs后,可能促進(jìn)SVZ區(qū)的內(nèi)源性NSCs向神經(jīng)元方向分化,減少向膠質(zhì)細(xì)胞方向分化。 3、移植NSCs后,可能改變了缺血區(qū)紋狀體的微環(huán)境,使得膠質(zhì)細(xì)胞增生的趨勢(shì)減弱。
[Abstract]:Objective: To observe the effect of Neural Stem Cells (NSCs) transplantation on the proliferation and differentiation of endogenous NSCs in the region of cerebral ischemia reperfusion rat and the endogenous NSCs in the subventricular zone of the cerebral ischemia reperfusion rat by establishing a rat model of cerebral ischemia reperfusion.
Methods: by extracting the hippocampus of fetal mice for 14 days, the rat model of NSCs. ischemic model was constructed by suspension culture for 24 hours. The rats were randomly divided into normal group, sham operation group, ischemic model group (hereinafter referred to as ischemic group) and ischemic transplant PBS group (below) under the guidance of brain stereotaxis guided by NSCs.. The PBS group and the ischemic transplanted neural stem cell group (hereinafter referred to as NSCs group) were observed on third days, 7 days and 14 days respectively. The assessment of ischemic nerve function loss caused by ischemia was evaluated using the evaluation criteria of 7 points, and the forelimb of all animals was detected by Foot-fault, and the changes of infarct area were observed by TTC. The morphology and structure of cells in the ischemic area were observed by Nissl staining. Apoptosis was detected by Tunel method and the number of apoptotic cells was counted. The expression of double standard positive cells in the striatum and subventricular zone (SVZ) was observed by Brdu/NeuN and Brdu/GFAP fluorescence double labeling method, and the number of positive cells in the related region was counted.
Result錛，

本文編號(hào)：1875372