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脂肪干细胞侧脑室定向移植改善脑出血大鼠的神经功能

Improvement of neural function by stereotaxic transplantation of adipose-derived stem cells into lateral cerebral ventricle after intracerebral hemorrhage in rats

摘要:

目的 研究脂肪干细胞(ADSC)侧脑室定向移植改善脑出血大鼠的神经功能及其作用机制.方法 体外培养大鼠ADSC,于移植前48 h用Brdu抗原标记.另取大鼠制备尾状核脑出血(ICH)模型,将建模后的大鼠分为2组,ADSC组于建模后48 h定向右侧脑室移植标记的ADSC,对照组输注等体积生理盐水.分别在完成ICH建模时及移植后1、3、7、14、28 d等时间点,对两组大鼠进行神经行为学评分,采用免疫荧光染色法检测Brdu阳性细胞及神经元和星形胶质细胞的分化情况,按照原位末端标记法(TUNEL)试剂盒的方法检测细胞凋亡情况,按照血管内皮细胞生长因子(VEGF)试剂盒说明检测VEGF的表达及血管新生情况.结果 体外ADSC具有分化为成脂肪细胞和成骨细胞的能力.在移植后3、7、14 d,ADSC组大鼠神经行为学评分明显优于对照组(P<0.05).免疫双荧光检测显示,Brdu标记的ADSC在大鼠脑组织内存活并且向病灶部位迁移,部分分化为神经胶质细胞和神经元细胞.TUNEL检测显示,移植后3 d时ADSC组凋亡细胞数目明显少于对照组(P<0.05).移植3 d后,ADSC组VEGF的表达均明显高于对照组(P<0.05).结论 经侧脑室定向移植的ADSC在脑损伤病灶周围可以存活,并可分化为神经样细胞;移植后ADSC可抑制细胞凋亡,分泌VEGF促进新生血管的生成,从而改善大鼠的神经功能.

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abstracts:

Objective To study improvement of neural function by stereotaxic transplantation of adipose-derived stem cells (ADSC) into lateral cerebral ventricle after intracerebral hemorrhage in rats and its mechanism. Methods ADSC were cultured and proliferated in vitro, which had been marked with Brdu for 48 h before transplantation. The rat caudate nucleus hemorrhage (ICH) models were divided into 2 groups. ADSC were stereotaxically transplanted into the right lateral ventricles in ADSC group, and equal volume of saline was transplanted into control group. The score of neurological behavior were evaluated at modeling and 1, 3, 7, 14, 28 days after transplantation respectively.Double-staining immunofluorescence technique was used to detect Brdu-positive cells and the differentiation of neurons and astrocytes. In accordance with the instructions of TUNEL kit, cell apoptosis, and the expression of VEGF and angiogenesis were assayed. Results In vitro ADSC expressed undergo osteogenic and adipogenic differentiation. Compared with the control group, ADSC group had better motor function at 3, 7, and 14 days (P<0. 05). Double-staining immunofluorescence showed mostly grafted Brdu-reactive ADSC had migrated to the hematoma zone, and some survivedand expressed Neun of Gfap. TUNEL analysis revealed that, 3 days after transplantation, the number of apoptotic cells in ADSC group was significantly less than in the control group (P<0. 05). Three days after transplantation, VEGF expression levels in ADSC group were significantly higher than in the control group (P<0. 05). Conclusion ADSC stereotaxially transplanted into the lateral ventricle can survive and differentiate into neuron-like cells. ADSC transplantation may reduce apoptosis and secret VEGF to promote the angiogenesis, and improve neural functional in intracerebral hemorrhage rats.

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作者: 陈娟 [1] 胡晓晴 [2] 谢雪微 [2] 刘永明 [2] 刘娜 [2] 曾文高 [2] 倪厚杰 [2] 王书新 [2] 骆翔 [2] 唐洲平 [2]
分类号: R74
栏目名称: 实验研究
DOI: 10.3760/cma.j.issn.0254-1785.2011.04.013
发布时间: 2011-06-10
基金项目:
国家自然科学基金 湖北省卫生厅科研重点项目
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