人羊膜间充质干细胞原位移植治疗大鼠脑梗死

发布时间：2018-09-09 11:56

【摘要】：背景:前期实验研究发现,移植人羊膜间充质干细胞能有效改善脑梗死大鼠的神经损伤症状。目的:观察移植人羊膜间充质干细胞在大鼠脑梗死区域的存活、定植和分化情况。方法:将60只SD大鼠随机分为3组,实验组、模型组采用线栓法制作大脑中动脉闭塞模型,假手术组只结扎血管,不插入线栓;造模后1 d,实验组于受损纹状体和皮质两点原位植入10μL第3代人羊膜间充质干细胞悬液(含细胞2×106/只),模型组与假手术组于相同部位注射等体积PBS。移植后1周连续监测体质量并进行神经缺损严重程度评分;移植后2周,TTC染色观察脑梗死面积,苏木精-伊红染色观察脑组织病理学变化,免疫荧光染色检测移植大鼠大脑神经细胞标志物神经元特异性核蛋白的表达。结果与结论:(1)体质量与神经缺损严重程度评分:与假手术组比较,模型组、实验组体质量呈下降趋势,实验组降低少于模型组,但差异无显著性意义;模型组、实验组神经功能缺损评分随时间呈逐渐降低趋势,但实验组评分明显低于模型组;(2)脑梗死面积:模型组大脑皮质出现局灶性缺血坏死且范围较大,与模型组比较,实验组缺血灶面积明显缩小;(3)脑组织病理:实验组梗死病灶范围、神经细胞及炎细胞浸润均少于模型组;(4)免疫荧光染色:移植后1周,实验组脑组织可见较多的移植人羊膜间充质干细胞,2周后可见移植的人羊膜间充质干细胞向神经样细胞分化;(5)结果表明:人羊膜间充质干细胞移植可在大鼠脑梗死区域定植、存活,且在原位能分化为神经样细胞。
[Abstract]:Background: transplantation of human amniotic mesenchymal stem cells (HMMSCs) can effectively improve the neurological injury symptoms in rats with cerebral infarction. Aim: to observe the survival, colonization and differentiation of transplanted human amniotic mesenchymal stem cells (HMMSCs) in cerebral infarction area of rats. Methods: sixty SD rats were randomly divided into three groups: the model group was made into middle cerebral artery occlusion model by thread occlusion, and the sham operation group only ligated the blood vessel without inserting the thread embolus. 1 day after modeling, the experimental group was implanted in situ with 10 渭 L human amniotic mesenchymal stem cell suspension (containing 2 脳 106 cells per mouse) at both the damaged striatum and cortex. The model group and the sham operation group were injected with PBS. of the same volume at the same site. One week after transplantation, the body mass was continuously monitored and the severity of nerve defect was evaluated, the area of cerebral infarction was observed by TTC staining and the pathological changes of brain tissue were observed by hematoxylin and eosin staining at 2 weeks after transplantation. The expression of neuron-specific nuclear protein was detected by immunofluorescence staining. Results and conclusion: (1) body mass and nerve defect severity score: compared with sham operation group, the body mass of model group, experimental group showed a downward trend, the experimental group decreased less than model group, but the difference was not significant, the model group, the model group, The neurological deficit score of the experimental group decreased gradually with time, but the score of the experimental group was obviously lower than that of the model group. (2) Cerebral infarction area: focal ischemic necrosis occurred in the cerebral cortex of the model group and the range was larger, compared with the model group. (3) brain histopathology: infarct area, nerve cell and inflammatory cell infiltration in experimental group were less than those in model group; (4) immunofluorescence staining: 1 week after transplantation, In the experimental group, more human amniotic mesenchymal stem cells could be found to differentiate into neural like cells 2 weeks after transplantation. (5) the results showed that the transplantation of human amniotic mesenchymal stem cells could be colonized in the infarcted area of rat brain. Survival, and in situ differentiation into neuronal cells.
【作者单位】：遵义医学院附属医院贵州省细胞工程重点实验室;遵义医学院药学院;遵义医学院附属医院临床医学研究所;遵义医学院教育部基础药理学重点实验室;
【基金】：贵州省科学技术基金项目资助(黔教研合J字LKZ[2011]24号)~~
【分类号】：R743.33

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