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目的观察粒细胞集落刺激因子(G-CSF)对移植的人神经干细胞(hNSCs)新生大鼠体内神经元分化的影响。方法出生1周的Wistar大鼠建立缺氧缺血性脑损伤(HIBD)模型,建模后分成2组:NSC移植组(N组,n=17)和NSC移植+G-CSF治疗组(NG组,n=16)。NG组大鼠建模后1h即皮下注射G-CSF,50μg.kg-1,1次.d-1,移植前后连续5d注射G-CSF。2组大鼠均于建模后第2天经脑室移植hNSCs。移植后1周、2周采用免疫荧光法观察2组植入细胞在皮质、海马部位神经元分化情况。结果移植后1周,分化神经元主要分布于大鼠损伤侧海马、皮质;移植后2周,神经元分化数量增多,并发出细长分枝。NG组海马各区神经元分化存活数量较N组多,差异有统计学意义(Pa<0.01);NG组皮质神经元分化存活数量在移植后2周时较N组多,差异有统计学意义(P<0.001)。结论 G-CSF有利于植入大鼠脑内的hNSCs的存活,并促进植入hNSCs向神经元分化,因此联合G-CSF可能有助于进一步提高hNSC移植的疗效。
Objective To observe the effects of G-CSF on the differentiation of neurons in neonatal rats with transplanted human neural stem cells (hNSCs). Methods One week old Wistar rats were divided into two groups: NSC transplantation group (n = 17) and NSC transplantation + G-CSF treatment group (n = 17) Group, n = 16). G-CSF was injected subcutaneously at 1 h after modeling in NG group, and 50 μg · kg -1, 1 d · d -1 were injected subcutaneously. G-CSF was injected 5 days after transplantation. Two groups of rats were transplanted transventricular hNSCs 2 days after modeling. Immunofluorescence was used to observe the differentiation of neurons in the cortex and hippocampus of the two groups after 1 week and 2 weeks after transplantation. Results At 1 week after transplantation, the differentiated neurons mainly distributed in hippocampus and cortex on the lesion side of rats. After 2 weeks of transplantation, the number of neuronal differentiation increased and slender branches emerged. The number of neuronal differentiation in hippocampus of NG group was more than that of N group (P <0.01). The number of survival of cortical neurons in NG group was more than that of N group two weeks after transplantation, the difference was statistically significant (P < P <0.001). Conclusion G-CSF is beneficial to the survival of hNSCs implanted in rat brain and promotes the differentiation of hNSCs into neurons. Therefore, the combination of G-CSF may help to further improve the efficacy of hNSC transplantation.