[1]陈太邦,赵建华,王永飞,等.体外树突状细胞通过NT-3上调P-ERK1/2的表达促进神经干/祖细胞分化[J].第三军医大学学报,2012,34(23):2368-2372.
 Chen Taibang,Zhao Jianhua,Wang Yongfei,et al.Dendritic cells promote neural stem/progenitor cells differentiation through NT-3 up-regulating p-ERK1/2 in vitro[J].J Third Mil Med Univ,2012,34(23):2368-2372.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
34卷
期数:
2012年第23期
页码:
2368-2372
栏目:
论著
出版日期:
2012-12-15

文章信息/Info

Title:
Dendritic cells promote neural stem/progenitor cells differentiation through NT-3 up-regulating p-ERK1/2 in vitro
作者:
陈太邦赵建华王永飞王钟郭乔楠
第三军医大学大坪医院野战外科研究所脊柱外科;第三军医大学西南医院病理学研究所
Author(s):
Chen Taibang Zhao Jianhua Wang Yongfei Wang Zhong Guo Qiaonan
Department of Spinal Surgery, Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing, 400042; Institute of Pathology, Southwest Hospital, Third Military Medical University, Chongqing, 400038, China
关键词:
干细胞树突状细胞共培养分化NT-3
Keywords:
stem cells dendritic cells co-culture differentiation NT-3
分类号:
R329.26; R329.28; R338.12
文献标志码:
A
摘要:
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。
Abstract:
Objective        To observe the effect of dendritic cells (DCs) on neural stem/progenitor cells (NSPCs) differentiation in vitro, and to explore the possible mechanism of neurotrophin-3 (NT-3) in NSPCs differentiation regulated by DCs.       Methods      Primary NSPCs and DCs generated from SD rats were separately cultured and then co-cultured by using transwell chambers, respectively. There are five experimental groups including a NSPCs group, a DCs group, a NSPCs/DCs group, a NSPCs+NT-3 group and a NSPCs/DCs+anti-NT-3 group. After incubation for 24, 48 and 72 h, ELISA was used to detect the NT-3 content in cell supernatant of each group. After 7 d, fluorescence immunocytochemistry was used to detect NSPCs differentiation, and Western blot assay was used to detect the expression of phosphorylated extracellular signal-regulated protein kinases (p-ERK1/2) of NSPCs.       Results       The ELISA results showed that the NT-3 content in the cell supernatant of the NSPCs/DCs group was significantly increased compared with the NSPCs group, DCs group and NSPCs/DCs+anti-NT-3 group (P<0.05). Seven days after induction of differentiation, the proportions of β-tubulin-Ⅲ-positive cells in the NSPCs/DCs group and NSPCs+NT-3 group were significantly higher than those of the NSPCs group and NSPCs/DCs+anti-NT-3 group (P<0.05), while the proportions of glial fibrillary acidic protein (GFAP)-positive cells in the NSPCs/DCs group and NSPCs+NT-3 group were significantly lower than those of the NSPCs group and NSPCs/DCs+anti-NT-3 group (P<0.05). The Western blotting results showed that the levels of p-ERK1/2 in the NSPCs/DCs group and NSPCs+NT-3 group were significantly higher than those in the NSPCs group, DCs group and NSPCs/DCs+anti-NT-3 group (P<0.05).       Conclusion       Co-culture of DCs and NSPCs can significantly promote NSPCs to differentiate into neurons. Activation of MEK-ERK signaling pathway may contribute to the differentiation of NSPCs into neurons, which is mediated by increased NT-3 binding to its specific receptor TrkC in co-culture system.

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更新日期/Last Update: 2012-12-04