论文部分内容阅读
背景:碱性成纤维细胞生长因子可以促进骨髓间充质干细胞的增殖和向神经细胞方向分化,并被认为是胶质细胞的分裂原。目的:以双重荧光标记验证静脉移植碱性成纤维细胞生长因子基因修饰的骨髓间充质干细胞在脑缺血模型大鼠脑内的存活及分化情况,及其向神经元样细胞和神经胶质细胞分化的趋势。设计、时间及地点:随机对照动物实验,于2005-07/2006-03在中南大学实验动物中心实验室完成。材料:选用50只SD大鼠,按随机数字表法分为4组:假手术组(n=10),脑缺血/再灌注损伤模型组(n=10),骨髓间充质干细胞治疗组(n=15),碱性成纤维细胞生长因子基因修饰的骨髓间充质干细胞治疗组(n=15)。方法:除假手术组外,其余3组制备局灶性脑缺血再灌注模型。分别将骨髓间充质干细胞或碱性成纤维细胞生长因子基因修饰的骨髓间充质干细胞通过静脉移植至实验性脑缺血大鼠体内,脑缺血再灌注损伤组大鼠注入相同体积的DMEM培养基。主要观察指标:应用5-溴-2-脱氧尿苷-神经元特异核蛋白及5-溴-2-脱氧尿苷-胶质纤维酸性蛋白双重荧光标记法观察移植细胞在脑内的存活和分化情况,比较各组大鼠脑缺血后的神经功能评分及脑梗死体积变化。结果:移植7d后,碱性成纤维细胞生长因子基因修饰的骨髓间充质干细胞组大鼠脑内5-溴-2-脱氧尿苷阳性细胞数、5-溴-2-脱氧尿苷-神经元特异核蛋白双标阳性细胞数均高于骨髓间充质干细胞治疗组(P<0.05),两组间5-溴-2-脱氧尿苷-胶质纤维酸性蛋白双标阳性细胞数差异无显著性意义(P>0.05)。再灌注7d后,静脉移植骨髓间充质干细胞和碱性成纤维细胞生长因子基因修饰的骨髓间充质干细胞均能改善脑缺血后大鼠的神经功能、减少脑梗死体积,碱性成纤维细胞生长因子基因修饰的骨髓间充质干细胞的作用明显优于骨髓间充质干细胞。结论:碱性成纤维细胞生长因子诱导的骨髓间充质干细胞静脉移植后可在脑内缺血区存活,并分化为比例更合适的神经元和神经胶质细胞,发挥神经修复作用。
BACKGROUND: Basic fibroblast growth factor (BfF) promotes the proliferation and differentiation of BMSCs into neurons and is considered as a mitogen for glial cells. OBJECTIVE: To verify the survival and differentiation of basic fibroblast growth factor-modified bone marrow-derived mesenchymal stem cells (MSCs) modified by vein transplantation in the brain of rat with cerebral ischemia model by double fluorescent labeling, The trend of cell differentiation. DESIGN, TIME AND SETTING: A randomized controlled animal experiment was performed at the Laboratory of Experimental Animal Center, Central South University between July 2005 and March 2006. MATERIALS: Fifty Sprague-Dawley rats were randomly divided into 4 groups: sham operation group (n = 10), cerebral ischemia / reperfusion injury model group (n = 10), bone marrow mesenchymal stem cell treatment group (n = 15) and basic fibroblast growth factor gene modified bone marrow mesenchymal stem cells (n = 15). Methods: In addition to sham operation group, the other three groups were given focal cerebral ischemia-reperfusion model. Bone marrow mesenchymal stem cells or basic fibroblast growth factor gene-modified bone marrow-derived mesenchymal stem cells were transplanted into the experimental cerebral ischemia rats by intravenous injection. The rats in the cerebral ischemia-reperfusion injury group were injected with the same volume of DMEM Medium. MAIN OUTCOME MEASURES: The survival and differentiation of transplanted cells in the brain were observed by dual fluorescent labeling with 5-bromo-2-deoxyuridine-neuron specific nucleoprotein and 5-bromo-2-deoxyuridine-glial fibrillary acidic protein The neurological deficit score and cerebral infarction volume of rats in each group were compared. Results: After 7 days of transplantation, the number of 5-bromo-2-deoxyuridine-positive cells, the number of 5-bromo-2-deoxyuridine-neurons in the brain of BMSCs modified with basic fibroblast growth factor The number of double-labeled positive cells of metabonomics was higher than that of bone marrow-derived mesenchymal stem cells (P <0.05). There was no significant difference between the two groups in the number of double-labeled positive cells of 5-bromo-2-deoxyuridine-glial fibrillary acidic protein Significant significance (P> 0.05). After 7 days of reperfusion, BMSCs and BMSC modified by BMSCs all improved the neurological function, decreased the volume of cerebral infarction, and increased the number of basic fibroblasts The role of cell growth factor gene modified bone marrow mesenchymal stem cells was significantly better than that of bone marrow mesenchymal stem cells. CONCLUSION: Basic fibroblast growth factor-induced bone marrow mesenchymal stem cells can survive in the ischemic area after cerebral vein transplantation and differentiate into more appropriate proportion neurons and glial cells, playing the role of nerve repair.