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目的将脑源性神经营养因子(BDNF)基因转入大鼠海马原代培养的神经干细胞(NSCs)中,获得 NSCs-BDNF 基因工程干细胞并移植治疗大鼠缺血性脑损伤。方法分离培养新生大鼠海马区 NSCs,检测其增殖、分化等特性;构建逆转录病毒载体 pLXSN-BDNF,转染 NSCs,获得NSCs-BDNF 基因工程干细胞,检测其 BDNF 的表达和活性;建立大鼠大脑中动脉局灶性脑缺血模型,通过立体定向技术将 NSCs-BDNF 移植入模型缺血侧海马,进行组织学和行为学检测。结果 NSCs-BDNF 移植后可以观察到动物行为学的改善,术后4周 Longa 评分1.343±0.293,脑切片可以观察到海马齿状回神经元数目的增加,术后4周存活率87.5%±6.6%,较对照有统计学意义(P<0.05)。结论 NSCs-BDNF 移植对实验性大鼠缺血性脑损伤有修复作用。
Objective To transfect BDNF gene into primary cultured neural stem cells (NSCs) of rat hippocampus and obtain NSCs-BDNF genetically engineered stem cells for transplantation for the treatment of ischemic brain injury in rats. Methods NSCs were isolated and cultured in hippocampus of neonatal rats to detect the proliferation and differentiation of NSCs. The retroviral vector pLXSN-BDNF was constructed and transfected into NSCs to obtain NSCs-BDNF genetically engineered stem cells. BDNF expression and activity were detected. Middle cerebral artery occlusion (MCAO) model was established. The NSCs-BDNF was transplanted into the ischemic lateral hippocampus via stereotactic technique for histological and behavioral tests. Results The animal behavioral improvement was observed after transplantation of NSCs-BDNF. Longa score was 1.343 ± 0.293 at 4 weeks after operation. The number of neurons in hippocampal gyrus was observed in brain slices and the survival rate at 4 weeks after operation was 87.5% ± 6.6 %, Compared with the control was statistically significant (P <0.05). Conclusion The transplantation of NSCs-BDNF can repair ischemic brain injury in experimental rats.