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目的观察树突状细胞(dendritic cells,DCs)对神经干/祖细胞(neural stem/progenitor cells,NSPCs)分化的影响,探讨神经营养素-3(Neurotrophin-3,NT-3)在DCs调控NSPCs分化中可能的作用机制。方法实验共设5组,分别为NSPCs组、NSPCs/DCs组、NSPCs+NT-3组、NSPCs/DCs+抗NT-3组和DCs组。共培养24、48、72 h后,ELISA法检测各组上清液中NT-3含量;7 d后荧光免疫细胞化学法检测各组NSPCs分化情况。Western blot检测各组NSPCs磷酸化的细胞外调节蛋白激酶(extracellular regulated protein kinases,ERK1/2)表达。结果 NSPCs/DCs组上清液中NT-3含量较NSPCs组、NSPCs/DCs+抗NT-3组和DCs显著升高(P<0.05)。NSPCs/DCs组和NSPCs+NT-3组β-tubulin-Ⅲ阳性细胞数较NSPCs组和NSPCs/DCs+抗NT-3组显著增高(P<0.05),而NSPCs组和NSPCs/DCs+抗NT-3组GFAP阳性细胞较NSPCs/DCs组和NSPCs+NT-3组显著增高(P<0.05)。NSPCs/DCs组和NSPCs+NT-3组NSPCs中的p-ERK1/2的表达较其余两组增高(P<0.05)。结论 NSPCs与DCs共培养能显著促进NSPCs分化为神经元,可能与共培养后NT-3表达量增高,通过其特异性受体TrkC激活下游信号通路MEK-ERK有关。
Objective To observe the effect of dendritic cells (DCs) on the differentiation of neural stem / progenitor cells (NSPCs) and to investigate the role of NT-3 in the regulation of NSPCs differentiation In the possible mechanism of action. Methods Five groups were divided into NSPCs group, NSPCs / DCs group, NSPCs + NT-3 group, NSPCs / DCs + anti-NT-3 group and DCs group. After co-cultured for 24, 48 and 72 h, the content of NT-3 in each supernatant was detected by ELISA. The differentiation of NSPCs in each group was detected by fluorescence immunocytochemistry after 7 d. Western blot was used to detect the phosphorylation of extracellular regulated protein kinases (ERK1 / 2) in NSPCs in each group. Results The level of NT-3 in NSPCs / DCs group was significantly higher than that in NSPCs group, NSPCs / DCs + anti-NT-3 group and DCs (P <0.05). The number of β-tubulin-Ⅲ positive cells in NSPCs / DCs group and NSPCs + NT-3 group was significantly higher than that in NSPCs group and NSPCs / DCs + NT-3 group (P <0.05) GFAP positive cells were significantly higher than NSPCs / DCs and NSPCs + NT-3 groups (P <0.05). The expression of p-ERK1 / 2 in NSPCs / DCs group and NSPCs + NT-3 group was higher than the other two groups (P <0.05). Conclusion Co-culture of NSPCs and DCs can significantly promote the differentiation of NSPCs into neurons, which may be related to the increased expression of NT-3 and the activation of downstream signaling pathway MEK-ERK through its specific receptor TrkC.