神经干细胞和多巴胺神经元联合移植对帕金森病大鼠运动障碍的影响

发布时间：2018-07-16 21:10

【摘要】：目的探讨神经干细胞和多巴胺神经元联合移植改善帕金森病大鼠运动障碍的效果。方法采用经典6-羟基多巴胺毁损法构建帕金森病大鼠模型36只,腹腔注射阿朴吗啡连续诱导4周。APO注射第4周将36只帕金森病大鼠随机分为对照组、多巴胺组、干细胞组及联合组,每组9只,分别于右侧纹状体区立体定向注入生理盐水、多巴胺神经元悬液、神经干细胞悬液、多巴胺神经元与神经干细胞悬液。计算各组移植后2、4、6、8周的旋转次数;干细胞组及联合组分别于移植后1、2、4、8周行MRI扫描,观察移植区影像学变化;移植后8周取各组脑组织行普鲁士蓝染色,观察移植细胞定植与迁移情况,免疫荧光法检测移植细胞分化情况。结果移植后2、4、6、8周,对照组、神经干细胞组、多巴胺组、联合组旋转次数依次降低,组间两两比较P均0.01。MRI检查结果:随着时间延长,联合组与干细胞组移植区T2 W/FFE成像上呈低信号影的区域逐渐变大,但联合组低信号影范围大于干细胞组。普鲁士蓝染色结果:干细胞组和联合组移植的神经干细胞及其分化后的细胞胞质内均可见大量蓝色颗粒,大部分存留在移植针孔附近,仅少部分细胞向远处迁移,但联合组蓝色颗粒数量以及扩散范围均大于干细胞组。联合组巢蛋白(Nestin)、酪氨酸羟化酶(TH)、绿色荧光蛋白(GFP)阳性细胞数及Nestin/GFP、TH/GFP均高于干细胞组,酸性纤维蛋白(GFAP)阳性细胞数及GFAP/GFP均低于干细胞组(P均0.01)。结论神经干细胞与多巴胺神经元联合移植可显著改善帕金森病大鼠的运动障碍,更有助于神经干细胞的定植、迁移及分化成为多巴胺神经元。
[Abstract]:Objective to investigate the effect of neural stem cell and dopamine neuron transplantation on motor disorder in Parkinson's disease rats. Methods Thirty-six rats with Parkinson's disease were induced by classical 6-hydroxydopamine damage method. 36 rats were randomly divided into control group and dopamine group by intraperitoneal injection of apomorphine for 4 weeks. Nine rats in each group were injected with normal saline, dopamine neuron suspension, neural stem cell suspension, dopamine neuron and neural stem cell suspension respectively in the right striatum. The rotation times of each group were calculated at 8 weeks after transplantation, MRI scanning was performed in the stem cell group and the combined group at the 1st and 2nd week after transplantation, and the imaging changes in the transplanted area were observed, and Prussian blue staining was performed in the brain tissue of each group at 8 weeks after transplantation. The colonization and migration of transplanted cells were observed, and the differentiation of transplanted cells was detected by immunofluorescence. Results the rotation times of the control group, the neural stem cell group, the dopamine group and the combined group decreased in turn at 8 weeks after transplantation (P < 0.01). The results of MRI showed that the rotation times of the control group, the neural stem cell group, the dopamine group and the combined group decreased in turn. The area of low signal intensity on T 2W / FFE images in the combined group and stem cell group gradually became larger, but the low signal range of the combined group was larger than that of the stem cell group. The results of Prussian blue staining showed that a large number of blue granules were found in the transplanted neural stem cells and their differentiated cytoplasm in stem cell group and combination group, most of them were located near the transplanted pinhole, and only a few of them migrated to the distance. However, the number of blue particles and the diffusion range in the combined group were larger than those in the stem cell group. The number of nestin, tyrosine hydroxylase (th), green fluorescent protein (GFP) positive cells and Nestin / GFP TH- / GFP in combination group were higher than those in stem cell group, and the number of acidic fibrin (GFAP) positive cells and GFAPP / GFP were lower than those in stem cell group (P0.01). Conclusion combined transplantation of neural stem cells and dopamine neurons can significantly improve the motor disorder of Parkinson's disease rats, and it is also helpful to the colonization, migration and differentiation of neural stem cells into dopamine neurons.
【作者单位】：昆明医科大学第一附属医院;
【基金】：国家自然科学基金资助项目(81241126,81360197) 云南省教育厅科学研究基金资助项目(2013C227) 云南省科技厅-昆明医学院联合专项(2014FB041)
【分类号】：R742.5

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