神经干细胞移植在脑缺血大鼠中的增殖与分化研究

发布时间：2018-05-11 19:46

  本文选题：神经干细胞 + 脑缺血　； 参考：《厦门大学》2014年硕士论文

【摘要】：目的：通过建立脑缺血再灌注大鼠模型,观察神经干细胞(Neural Stem Cells, NSCs)移植对脑缺血再灌注大鼠纹状体梗死区域新生细胞以及侧脑室下区内源性NSCs增殖分化的影响。 方法：通过提取孕14天胎鼠海马,利用悬浮培养法分离培养鼠源性NSCs。缺血模型构建成功后24小时,在脑立体定位仪引导下向左侧纹状体注射NSCs。将实验大鼠随机分为正常组、假手术组、缺血模型组(以下简称缺血组)、缺血移植PBS组(以下简称PBS组)和缺血移植神经干细胞组(以下简称NSCs组),分别于第3天、7天、14天进行观察。利用7分制的评估标准对缺血引起的神经功能缺失状况进行评估；采用Foot-fault的方法对所有动物损伤的前肢进行了检测；TTC观察梗塞面积的变化;尼氏染色结果观察缺血区细胞形态和结构；Tunel法检测细胞凋亡并进行凋亡阳性细胞计数；Brdu/NeuN、Brdu/GFAP荧光双标法观察纹状体以及侧脑室下区(SVZ)双标阳性细胞的表达情况；统计相关区域阳性细胞数。 结果： 1、NSCs组较PBS组的右侧肢体偏瘫程度减轻。 2、TTC染色：NSCs组较PBS组的脑梗死面积明显减小。 3、尼氏染色：PBS组的缺血区有大量的神经元坏死,损伤主要集中在纹状体,NSCs组神经细胞的变性损伤范围明显减少。 4、Tunel(细胞凋亡染色)：NSCs组在各时间点的凋亡阳性细胞数均少于PBS组。 5、免疫荧光双标：SVZ区内源性神经干细胞在第3天开始出现增殖,第7天到达最高峰,第14天明显减少。其中,NSCs组较PBS组的Brdu/NeuN数目明显增多,差异有统计学意义(P0.05)。NSCs组较PBS组的Brdu/GFAP数目明显减少,差异有统计学意义(P0.05)。PBS组纹状体可见大量Brdu/GFAP阳性细胞,而NSCs组Brdu/GFAP阳性细胞较少。 结论： 1、移植NSCs后,神经功能缺失症状以及协调能力得到了明显的改善。 2、移植NSCs后,可能促进SVZ区的内源性NSCs向神经元方向分化,减少向胶质细胞方向分化。 3、移植NSCs后,可能改变了缺血区纹状体的微环境,使得胶质细胞增生的趋势减弱。
[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锛，

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