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目的探讨高海拔地区大鼠骨髓源神经干细胞(bone marrow mesenchymal stem cells-derived neural stem cells,BMSCs-NSCs)及脑源性神经生长因子(Brain-derived neurotrophic factor,BDNF)联合移植对大鼠脑缺血再灌注模型的疗效及其相关机理。方法 60只Wistar雄性大鼠,置西宁地区正常饲养,制备大鼠脑缺血-再灌注损伤模型;模型制备完毕后立体定向下进行细胞移植治疗,将大鼠分为3组,即A组:大鼠骨髓源性神经球组(BMSCs-NSCs组,n=20);B组:大鼠骨髓源性神经球联合BDNF组(BMSCs-NSCs+BDNF组,n=20),注射大鼠骨髓源性神经球细胞的同时,联合注射100ng BNDF;C组:对照组(仅注射DMEM/F12培养基,n=20);术后对其神经功能进行评定,并于术后24d取脑组织,行Nestin、GFAP、Map2免疫荧光检测。结果细胞移植后第3d,各组间神经功能评分无显著性差异;细胞移植后第14dBMSCs-NSCs+BDNF组神经功能评分显著优于BMSCs-NSCs组,BMSCs-NSCs组优于对照组;免疫组化检测发现,BMSCs-NSCS+BDNF组Nestin、GFAP、Map2的IOD值均显著高于BMSCs-NSCs组;BMSCs-NSCs+BDNF组、BMSCsNSCs组各检测指标水平均高于对照组;Nestin、GFAP、Map2的表达主要集聚于脑梗死灶与正常脑组织交界处。结论在西宁地区联合移植大鼠骨髓源神经干细胞及BDNF可显著促进大鼠大脑中动脉闭塞再灌注损伤模型的神经功能恢复。
Objective To investigate the effects of transplantation of bone marrow mesenchymal stem cell-derived neural stem cells (BMSCs-NSCs) and brain-derived neurotrophic factor (BDNF) Therapeutic effect of blood reperfusion model and its related mechanism. Methods Sixty male Wistar rats were housed in Xining area to prepare a model of cerebral ischemia-reperfusion injury in rats. After the preparation of the model, the cells were transplanted stereotacticly. The rats were divided into three groups: Group A: BMSCs-NSCs group (n = 20); B group: Bone marrow-derived neurospheres combined with BDNF group (BMSCs-NSCs + BDNF group, n = 20) 100ng of BNDF were injected simultaneously in group B and group B; Group C: control group (only DMEM / F12 medium was injected, n = 20); neurological function was evaluated after operation and brain tissue was taken 24 days after operation Nestin, GFAP, Map2 immunofluorescence assay. Results The neural function score of the 14th day after transplantation was significantly higher than that of the BMSCs-NSCs group (P <0.05), while the BMSCs-NSCs group was superior to the control group The IOD values of Nestin, GFAP and Map2 in BMSCs-NSCS + BDNF group were significantly higher than that in BMSCs-NSCs group; the levels of BMSCs-NSCs + BDNF group and BMSCs NSCs group were higher than those in control group; Nestin, GFAP, Map2 expression mainly concentrated in the cerebral infarction and normal brain tissue junction. Conclusion Co-transplantation of rat bone marrow-derived neural stem cells and BDNF in Xining area can significantly promote the recovery of neural function in the rat model of middle cerebral artery occlusion-reperfusion injury.