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背景:帕金森病是一种常见的神经系统变性疾病,胚胎干细胞移植治疗能缓解帕金森病的症状,但受技术和伦理学方面制约。与胚胎干细胞比较,骨髓基质多能成体祖细胞的许多特性也使其逐渐成为细胞移植治疗中理想的细胞资源之一。目的:探讨骨髓基质分离的多能成体祖细胞通过系统移植方式进入大鼠脑组织内并修复受损的神经功能。设计:随机对照实验。单位:武汉市第一医院神经内科。材料:实验于2003-10/2005-03在华中科技大学同济医学院附属协和医院神经内科完成。选用健康SD大鼠80只,体质量180～200g,由华中科技大学同济医学院实验动物中心提供。方法:制作实验性帕金森病大鼠模型,将在体外纯化、增殖和已用5-溴-2-脱氧尿苷处理过的骨髓基质多能成体祖细胞通过尾静脉注入帕金森病大鼠体内。3个月后,对移植大鼠采用免疫组化技术、RT-PCR、免疫电镜、行为学评定等方法鉴定移行入大鼠脑组织内的骨髓基质多能成体祖细胞、其分化的神经元样细胞以及神经功能的修复。主要观察指标:①行为学观察结果。②免疫组织化学染色结果。结果:骨髓基质多能成体祖细胞能移行入大鼠脑组织内并在中脑黑质和纹状体区分化为神经元样细胞;6-羟多巴诱导的大鼠行为损伤有明显恢复;多巴胺-β-羟化酶、神经生长因子和多巴胺转运体mRNA的表达水平明显升高;电镜下观察到骨髓基质多能成体祖细胞所分化的神经细胞与其它神经细胞形成突触联系。结论:骨髓基质多能成体祖细胞能在大鼠脑组织内分化为多巴胺能神经细胞并发挥相应的神经功能。
BACKGROUND: Parkinson’s disease is a common degenerative disease of the nervous system. Embryonic stem cell transplantation can relieve the symptoms of Parkinson’s disease but is restricted by technical and ethical aspects. Compared with embryonic stem cells, many characteristics of marrow stromal multipotent adult progenitor cells also make it gradually become one of the ideal cell resources in cell transplantation therapy. OBJECTIVE: To investigate whether multipotent adult progenitor cells isolated from bone marrow stromal cells enter the rat brain through systemic transplantation and repair impaired neurological function. Design: Randomized controlled experiment. Unit: First Hospital of Wuhan Neurology. MATERIALS: The experiment was performed at Department of Neurology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from October 2003 to March 2005. Eighty healthy SD rats were selected and their body weight was 180 ~ 200g. They were provided by Laboratory Animal Center, Tongji Medical College, Huazhong University of Science and Technology. METHODS: A rat model of experimental Parkinson’s disease was established, and in vitro purified, proliferated and injected with bone marrow stromal multipotent adult progenitor cells that had been treated with 5-bromo-2-deoxyuridine into Parkinson’s disease rats . Three months later, the transplanted rats were identified by immunohistochemistry, RT-PCR, immunoelectron microscopy, behavioral assessment and other methods to migrate into the brain tissue of rat bone marrow stromal multipotent progenitor cells, differentiated neuron-like Cell and nerve function repair. MAIN OUTCOME MEASURES: ①Behavioral observation results. ② immunohistochemical staining results. RESULTS: Bone marrow stromal multipotent adult progenitor cells migrated into rat brain tissue and differentiated into neuron-like cells in substantia nigra and striatum of midbrain; the behavioral injury induced by 6-hydroxydopa was significantly restored; The expression levels of dopamine-β-hydroxylase, nerve growth factor and dopamine transporter mRNA were significantly increased. The synaptic connections between neural cells differentiated from the multipotent adult progenitor cells of bone marrow stroma and other neurons were observed under electron microscope. CONCLUSION: Bone marrow stromal multipotent adult progenitor cells can differentiate into dopaminergic neurons in rat brain and exert corresponding neurological functions.