骨髓基质多能成体祖细胞移植治疗大鼠帕金森病(英文)

Transplantation of bone marrow multipotent adult progenitor cells for treating Parkinson disease in rats

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 disease is a common degenerative disorder of nervous system. Transplantation of embryonic stem cell can alleviate the symptoms of Parkinson disease, but restdcted technically and ethically. Compared with embryonic stem cell, the various characteristics of bone marrow dedved-multipotent adult progenitor cells （MAPCs） enable them to become one the ideal sources of cells for cell transplantation. OBJECTIVE： To explore the hypothesis that MAPCs ware able to enter the brain and reduce the neurological functional defidlts in rats by injecting intravenously. DESIGN ： A randomized controlled experiment. SETTING： Department of Neurology, Wuhan First Hospital MATERIALS ： The experiments ware performed in the laboratory of Department of Neurology, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology from October 2003 to March 2005. Eighty healthy Sprague-Dawley （SD） rats of 180-200 g ware provided by the experimental animal center of Tongji Medical College, Huazhong University of Science and Technology. METHODS： The rats ware made into models of Parkinson disease, the bone marrow-derived MAPCs, which ware in vitro purified, proliferated and treated with 5-bromo-2-deoxyufidine （BrdU）, ware injected via caudal vein. After three months, the immunohistochemical technique, reverse transcription-polymerase chain reaction （RT-PCR）, electron microscopy and behavioral tests ware used to identify the MAPCs or neuron-like cells derived from MAPCs in brain and their functions. MAIN OUTCOME MEASURES： （1） Results of behavioral observation; （2） Results of immunihistochemical staining RESULTS： After implantation, MAPCs could survive and differentiate into neuron-like cells in substantia nigra and stdatum. MAPCs-derived dopaminergic neurons caused gradual and sustained behavioral restoration of 6-hydroxydopamine （6-OHDA）-mediated motor asymmetry. The levels of dopamine beta-hydroxylase （DBH）, nerve growth factor （NGF） or dopamine transporter （DAT） mRNA ware up-regulated significantly. It was observed under electron microscope that immature synapse implicated MAPCs- derived neuron should play an important role in the reconstruction of neural circuitry. CONCLUSION： Transplanted bone marrow dedved-MAPCs can spontaneously differentiate into dopaminergic neurons, and act the corresponding nerve function.