目的:研究鼠神经干细胞(NSCs)移植治疗脑缺血大鼠的效果及清热化瘀方的影响.方法:分离、培养新生大鼠海马NSCs,采用线栓法制作大鼠脑缺血模型后经侧脑室移植未分化的NSCs入脑缺血大鼠脑内,对移植后大鼠进行神经损害严重程度评分( NSS),用免疫组化(荧光)方法观察移植后NSCs的存活、迁徙、分化状况.结果:大鼠脑缺血损伤可以诱导少量nestin的表达,且在缺血再灌注7d时表达呈高峰(P＜0.05)；经BrdU标记的NSCs侧脑室移植后2d即可见BrdU蛋白阳性细胞,并可见其从室旁区向缺血区移行,其表达于移植后14d达高峰(P＜0.05)；免疫荧光显示,NSCs移植后7d可见BrdU/NF200双阳性细胞出现在缺血周围处,其表达随缺血时间延长而逐渐增加；移植加中药组可显著增加缺血后第14、28天时缺血半暗带区nestin、BrdU和BrdU/NF200阳性蛋白表达(P＜0.01,P＜0.05),并改善缺血后第7、14、28天的NSS(P ＜0.05).结论:联合应用清热化瘀方可促进NSCs增殖并分化为神经元,促进神经功能的恢复.%Objective:To examine the effect of neural stem cells(NSCs) transplantation on rats with cerebral ischemia injury and the role of Qingre Huayu Prescription (QRHYP). Methods: Hippocampus NSCs were isolated and cultured in vitro, which were injected into lateral ventricle in rat models of brain ischemia established by middle cerebral artery occlusion( MCAO). The neurological deficit scores(NSS) were determind and immunohistochcmistry ( fluorescence) were used to check the differentiation and migration of NSCs. Results;The expression of nestin protein ischemic region at 7d after NSCs transplantation,whose expression increased continuously with the extension of ischemic time. The positive proteins expression of nestin,BrdU and BrdU/NF200 in NSCs transplantation + QRHYP group were significantly higher than those of in simple transplantation group at 14 and 28d ( P < 0. 01 , P < 0. 05 ) . Compared with the model group, NSS of NSCs transplantation + QRHYP group were obviously better at 7d,14d and 28d ( P < 0. 05 ) . Conclusions:The transplantation of NSCs combined with QRHYP can stimulate NSCs proliferation and differentiation into neurons, promote the recovery of neurological function following ischemia brain damage.
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