【作者】 刘洋；

【导师】 薛洪利；

【作者基本信息】 大连医科大学 ， 外科学， 2008， 硕士

【摘要】 干细胞移植治疗已经成为脑外伤及部分神经系统疾病最有潜力和发展前途的治疗手段。骨髓间充质干细胞(mesenchymal stem cells,MSCs)由于其来源广泛、便于体外扩增而成为最佳的干细胞来源。间充质干细胞既可以从人体骨髓组织中直接获得,也可以通过骨髓动员,使骨髓干细胞大量释放入血,经过体外循环分离获得。移植入中枢神经系统后,MSCs可以分化为神经元,神经胶质细胞及其他的主要神经系统细胞。目前认为,MSCs的分化潜能、MSCs及损伤脑组织分泌的细胞因子的相互作用是干细胞分化为神经系统细胞的主要影响因素。研究MSCs在损伤脑组织内的分化机制对间充质干细胞移植治疗具有决定性的意义。第一部分颈内动脉注射自体骨髓MSCs移植治疗重型颅脑伤后遗症的动物实验目的探索自体骨髓间充质干细胞对脑损伤的治疗作用。方法将Wistar大鼠自体骨髓MSCs在体外扩增并经Brdu标记后,通过颈内动脉注射将其植入冷冻伤脑水肿模型体内,从组织化学和神经功能评分两个方面观察骨髓MSCs自体移植的治疗作用。结果经颈内动脉注射的Brdu标记的大鼠MSCs细胞,可向脑损伤区域迁移。B组Brdu阳性细胞的数量和神经功能恢复的程度均高于A组,且有明显统计学差异(P<0.05)。与B组相比C组使用神经节苷脂和丹参注射液未增加Brdu阳性细胞的数量,但是神经功能恢复的程度明显升高(P<0.05)。D组使用罂粟碱开放血脑屏障,Brdu阳性细胞的数量,神经功能恢复的程度均高于B组。但神经恢复的程度与C组相比无明显差异(P>0.05)。结论骨髓间充质干细胞可以通过循环系统向脑损伤区迁移而发挥治疗作用。第二部分自体骨髓间充质干细胞不同移植途径对大鼠神经功能改善的影响目的探索自体骨髓间充质干细胞不同移植途径对大鼠神经功能改善的影响。方法将Wistar大鼠自体骨髓间充质干细胞在体外扩增并经Brdu标记后,通过颈内动脉和脑室系统注射将其植入冷冻伤脑水肿模型体内,从组织化学和神经功能评分两个方面观察骨髓MSCs自体移植的治疗作用。结果经颈内动脉和脑室系统注射的Brdu标记的大鼠MSCs细胞,可向脑损伤区域迁移。B组和C组Brdu阳性细胞的数量和神经功能恢复的程度均高于A组,且有明显统计学差异(P<0.05)。B组和C组Brdu阳性细胞的数量和神经功能恢复的程度相比无明显统计学差异(P>0.05)。结论骨髓间充质干细胞可以通过循环系统和脑室系统向脑损伤区迁移而发挥治疗作用,二者无统计学差异。更多还原

【Abstract】 Stem cells transplantation have great potential as a therapeutic tool for the repair of cerebral trauma and a number of other CNS disorders. Because the source of the cells is extensive and it can proliferate a great quantity in vitro, MSCs(marrow-derived mesenchymal stem cells,MSCs) will be the best choice for transplantation. MSCs can either be isolated from human bone marrow directly or through extracorporeal circulation when stem cells release into the blood after bone marrow mobilization. It is considered that the multipotentiality of MSCs and the interaction of the MSCs and the cytokines from MSCs and the injuered brain tissue are the main influencing factors that MSCs differentiates into main cells in CNS. Following engraftment into the adult CNS, MSCs differentiate mainly into neuron,glia and other main cells in CNS. Understanding the mechanisms of MSCs differentiation in the context of the injured CNS will be critical to achieving these therapeutic strategies.PartI The Treatment Action of MSCs Auto-transplatation on the Severe Brain Injury through the internal carotid artery .Objective: To explore the action of bone mesenchymal stem cells (MSCs) in treatment of brain injury following MSCs auto trans-plantation. Methods:After MSCs from the Wistars were proliferated,marked with Brdu in vitro,the Brdu-marked MSCs were transplanted into the cold-induced brain edema model through the internal carotid artery . We observed the treatment of MSCs through histochemistry and evaluation of the neural function.Results: The Brdu-marked MSCs from the Wistars can immigrate into the brain injury area. The amount of Brdu positive cells in the damaged area and the points of the neural functiong in the auto-transplatation groups(group B) were larger than the control group(group A),with significant statistical difference (P<0.05). The amount of Brdu positive cells in the group C with GM1 AND Dansen, injection were not larger than group B,but the points of the neural functiong were higher significantly.Group D used cardoverine to open the BBB, the amount of Brdu positive cells were larger than group B and C, the points of the neural functiong were larger than group B, with no significant statistical difference compared with group C (P>0.05).Conclusion: The MSCs marked with Brdu can immigrate to the injury area through the internal carotid artery and exert the therapeutic action.PartII The Treatment Action of MSCs Auto-transplatation on the Severe Brain Injury through different transplatation pathwaysObjective: To explore the action of bone mesenchymal stem cells (MSCs) in treatment of brain injury following MSCs auto-transplant-ation through different transplantation pathways.Methods:After MSCs from the Wistars were proliferated,marked with Brdu in vitro,the Brdu-marked MSCs were transplanted into the cold-induced brain edema model through the internal carotid artery and the ventricular system . We observed the treatment of MSCs through histochemistry and evaluation of the neural function.Results: The Brdu-marked MSCs from the Wistars can immigrate into the brain injury area through the internal carotid artery and the ventricular system. The amount of Brdu positive cells in the damaged area and the points of the neural functiong in the auto-transplatation groups(group B and C) were larger than the control group(group A),with significant statistical difference (P<0.05). The amount of Brdu positive cells and the points of the neural functiong in the group B and C have no significant statistical difference (P>0.05).Conclusion: The MSCs marked with Brdu can immigrate through the internal carotid artery and the ventricular system to the injury area and exert the therapeutic action,with no significant statistical difference.更多还原