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目的观察缺氧缺血脑损伤(hypoxic-ischemic brain damage,HIBD)新生大鼠应用粒细胞集落刺激因子(granulocyte-colony stimulating factor,G-CSF)对移植的人神经干细胞(human neural stem cells,hNSCs)体内存活及分化成少突胶质细胞的影响。方法新生1周的Wistar大鼠HIBD模型后分成两组:NSCs移植组(A组,n=24)和NSCs移植+G-CSF治疗组(B组,n=24),其中B组建模后1h即开始皮下注射G-CSF,每天1次,移植前后连续5d注射G-CSF。两组均于建模后第2天经脑室移植hNSCs。移植后3d用TUNEL法检测植入细胞凋亡情况,并于移植后1周、2周分别用免疫荧光法检测植入细胞在大鼠脑内的存活及向少突胶质细胞方向分化情况。结果移植后3d,A组大脑皮层见大量的植入细胞处于凋亡状态,而B组的植入细胞凋亡明显减少(t=68.36,P<0.01)。移植后1周及2周,可见植入细胞分布以额叶皮层和海马多见,纹状体亦有少许植入细胞分布,A组的植入细胞数少于B组(t=14.07和2.5,P<0.05)。而移植后1周及2周,hNSCs几乎不能分化成少突胶质细胞。结论 G-CSF能减少植入细胞的凋亡,提高植入细胞的存活率,但对植入细胞向少突胶质细胞方向的分化无影响。
Objective To observe the effect of granulocyte-colony stimulating factor (G-CSF) on human neural stem cells (hNSCs) in neonatal rats with hypoxic-ischemic brain damage (HIBD) ) In vivo survival and differentiation into oligodendrocytes. Methods The newborn Wistar rats were divided into two groups: the NSCs transplantation group (group A, n = 24) and the NSCs transplantation + G-CSF treatment group (group B, n = 24) 1h began to subcutaneous G-CSF, once a day, 5d consecutive days before and after G-CSF injection. Both groups were transplanted transventricular hNSCs 2 days after modeling. Three days after transplantation, the apoptosis of implanted cells was detected by TUNEL method. Survival and differentiation of the implanted cells in the rat brain were detected by immunofluorescence at 1 and 2 weeks after transplantation. Results Three days after transplantation, a large number of implanted cells in the cerebral cortex of group A were found to be in apoptotic state, but apoptosis in group B was significantly reduced (t = 68.36, P <0.01). At 1 week and 2 weeks after transplantation, the distribution of implanted cells was more common in the frontal cortex and hippocampus, and there were a few implanted cells in the striatum. The number of implanted cells in group A was less than that in group B (t = 14.07 and 2.5 , P <0.05). At 1 week and 2 weeks after transplantation, hNSCs hardly differentiated into oligodendrocytes. Conclusion G-CSF can reduce the apoptosis of implanted cells and increase the survival rate of implanted cells. However, G-CSF has no effect on the differentiation of implanted cells into oligodendrocytes.