大鼠骨髓间质多潜能成年祖细胞体内移植与多巴胺能神经元分化的研究

Bone marrow mulupotent adult progenitor cells develop into functional dopaminergic neurons after transplantation in Parkinson rat models

目的:探讨骨髓间质分离的多潜能成年祖细胞(MAPCs)通过系统移植(即尾静脉注射)方式进入大鼠脑组织内并修复受损的神经功能。方法:制作实验性帕金森疾病大鼠模型.将在体外纯化、增殖和已用5-溴-2脱氧尿苷(BrdUrd)处理过的多潜能成年祖细胞通过尾静脉注射入帕金森病大鼠体内。三个月后,对受试大鼠进行行为学评定;并应用免疫荧光化学技术和RT-PCR等方法对脑组织内的MAPCs及其分化细胞进行鉴定。结果:多潜能成年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.如多巴胺能神经元,免疫荧光染色显示5-溴-2脱氧尿苷、神经元特异性烯醇化酶(NSE)或酪胺酸羟化酶(TH)表达阳性;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(DBH)和多巴胺转运体(DAT)mRNA的表达水平明显升高。结论:骨髓间质分离的多潜能成年祖细胞能通过系统移植方式进入大鼠脑组织内,在中脑微环境中可自主分化为多巴胺能神经细胞并有效地修复6-羟多巴诱导的神经功能缺损。因此,多潜能成年祖细胞有望成为中枢神经系统疾病自体移植治疗的最佳候选干细胞之一。

Objective: We tested the hypothesis that bone marrow derived- multipotent adult progenitor cells (MAPCs) enter the brain and reduce neurological functional deficits in rats by injecting intravenously. Methods: We establish successfully animal models of parkinson's disease in rats, sequentially, we injected MAPCs by tail vein, for cellular identification, MAPCs were prelabeled with bromodeoxyuridine(BrdUrd). By three months postinjection, behavioral tests - immunofluorescence method and RT-PCR were used to identify MAPCs or neuron-like cells derived from MAPCs in brain tissues. Results: After implantation, MAPCs could survive and differentiate into neuron-kike cells in substantia nigra and striatum, including dopamine-neurons. Compared with control animals, MAPCs-derived DA neurons caused gradual and sustained behavioral restoration of DA-mediated motor asymmetry. Levels of DBH and DAT mRNA were upregulated significantly. Conclusions: These results demonstrate that transplanted MAPCs can develop spontaneously into DA neurons. Such DA neurons can restore cerebral function and behavior in rat models of Parkinson's disease, MAPCs may provide a powerful autoplastic therapy for a variety of central nervous system disorders.