Notch1在全反式维甲酸诱导的神经干细胞分化中的作用

doi:10.6040/j.issn.1671-7554.0.2017.1194

摘要/Abstract

摘要： 目的探究Notch1在全反式维甲酸诱导的神经干细胞分化中的作用,揭示其对神经管发育的影响。方法免疫荧光检测神经干细胞标记物——巢蛋白(Nestin)、神经元标记物——神经丝蛋白(NF)、星型胶质细胞标记物——神经胶质纤维酸性蛋白(GFAP)及少突胶质细胞标记物——半乳糖脑苷脂(GALC)及Notch1在上述细胞中的分布。Western blotting检测神经干细胞中Notch1、NF、GFAP、GALC分别在普通分化培养基和全反式维甲酸培养基(ATRA)中1、3、5、7 d的含量。神经干细胞分为4组:将普通分化培养基设为A组,普通分化培养基+DMSO组设为B组,普通分化培养基+DMSO+25 μmol/L γ-内分泌酶抑制剂(γSI)组设为C组,普通分化培养基+DMSO+50 μmol/L γSI设为D组,Western blotting检测各组细胞中Notch1含量。结果免疫荧光鉴定Nestin、NF、GFAP、GALC呈阳性,且在上述细胞胞膜上及胞质中有Notch1存在。普通分化培养基中Notch1在第5天较第1天明显增加,NF在第5天较第3天明显降低,GFAP与GALC含量在第5天较第1天明显下降。加入ATRA的普通分化培养基中,Notch1在第7天较第1天明显减少,而NF、GFAP、GALC在第7天较第1天明显增加。加入γSI后,Notch1胞内段NICD含量明显降低。结论 Notch1信号通路抑制神经干细胞分化,ATRA通过抑制Notch1促进神经干细胞的分化。

关键词: 神经管缺陷, 全反式维甲酸, Notch1, 神经干细胞

Abstract: Objective To investigate the role of Notch1 in neural stem cell differentiation induced by all-trans retinoic acid and to reveal its effects on neural tube development. Methods The expressions of Notch1 in nestin, neurofilament(NF), glial fibrillary acidic protein(GFAP)and galactocerebroside(GALC)were identified with immunofluorescence. The neural stem cells were cultured in normal differentiation medium and the all-trans retinoic acid(ATRA)medium. The expressions of nestin, NF, GFAP and GALC on day 1, 3, 5 and 7 were detected with Western blotting. The neural stem cells were divided into 4 groups and treated with normal differentiation medium(group A), normal differentiation medium + DMSO(group B), normal differentiation media + DMSO + 25 μmol/L γ-secretase inhibitor(group C), and normal differentiation media + DMSO + 50 μmol/L γ-secretase inhibitors(group D). The expressions of Notch1 in these groups were detected with Western blotting. Results Nestin, NF, GFAP and GALC were positively expressed, and Notch1 distributed in the cell membrane and cytoplasm. In the normal differentiation medium, Notch1 山东大学学报 (医学版)56卷10期 -徐加明,等.Notch1在全反式维甲酸诱导的神经干细胞分化中的作用 \=-significantly increased on day 5 compared with day 1, NF decreased significantly on day 5 compared with day 3, GFAP and GALC significantly decreased on day 5 compared with day 1. In the ATRA medium, Notch1 significantly reduced on day 7 compared with day 1, whereas NF, GFAP, and GALC significantly increased on day 7. In after adoling the γSI, the intracellular domain(NICD)of Notch1 decreased significantly. Conclusion Notch1 signal pathway inhibits neural stem cell differentiation, while ATRA promotes neural stem cell differentiation by inhibiting Notch1.

Key words: Neural tube defect, All-trans retinoic acid, Notch1, Neural stem cell

中图分类号:

R729

徐加明,马爱华,陈娜,韩波. Notch1在全反式维甲酸诱导的神经干细胞分化中的作用[J]. 山东大学学报 (医学版), 2018, 56(10): 72-78.

XU Jiaming, MA Aihua, CHEN Na, HAN Bo. Effects of Notch1 on neural stem cell differentiation induced by all-trans retinoic acid[J]. Journal of Shandong University (Health Sciences), 2018, 56(10): 72-78.

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