Role of Notch - 1 in the Differentiation of Rat Mesenchymal Stem Cells into Neurons

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Role of Notch - 1 in the Differentiation of Rat Mesenchymal Stem Cells into Neurons

更新时间：2024-01-17

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- Role of Notch - 1 in the Differentiation of Rat Mesenchymal Stem Cells into Neurons
- Journal of Chongqing Medical University Issue 6, Pages: 781-786(2005)
- 作者机构：
  
  1. 重庆医科大学附属第一医院神经内科
  2. 郑州大学附属第一医院神经内科
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2005.06.003
  CLC： R329.2
- Published：2005，
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[1]杨军,贾延劼,杨琴,承欧梅.Notch-1在骨髓间质干细胞分化为神经细胞中的作用[J].重庆医科大学学报,2005(06):781-786.

YANG Jun, et al. Role of Notch - 1 in the Differentiation of Rat Mesenchymal Stem Cells into Neurons. [J]. Journal of Chongqing Medical University (6):781-786(2005)
[1]杨军,贾延劼,杨琴,承欧梅.Notch-1在骨髓间质干细胞分化为神经细胞中的作用[J].重庆医科大学学报,2005(06):781-786. DOI： 10.13406/j.cnki.cyxb.2005.06.003.

YANG Jun, et al. Role of Notch - 1 in the Differentiation of Rat Mesenchymal Stem Cells into Neurons. [J]. Journal of Chongqing Medical University (6):781-786(2005) DOI： 10.13406/j.cnki.cyxb.2005.06.003.

摘要

目的:采用RNA干扰（RNA interference

RNAi）技术

抑制Notch-1基因的表达

从而观察Notch-1基因在骨髓间质干细胞（MSCs）分化为神经细胞后的存活及分化中的作用。方法:构建mNotch-1短发夹状RNA（short hairpin RNA

mNotch -1 shRNA）;然后对mNotch-lshRNA转染的MSCs进行形态学观察;MTT检测转染前、转染24h、转染3天的MSCs的存活率; 体外条件下采用BDNF、全反式维甲酸等诱导mNotch-lshRNA转染的MSCs分化

采用免疫细胞化学染色

检测Nestin、NSE 及NF200（神经元标记物）、GFAP（神经胶质细胞标记物）的表达。结果:mNotch-lshRNA转染成功的细胞立体感增强

Notch -1基因表达消失

转染后24h和3天的MSCs细胞存活率下降

与未转染和转染对照组比较有显著性差异（P<0．01）;转染 mNotch-lshRNA的MSCs向神经细胞的分化效率显著提高

NSE、NF200的表达均高于未转染的和转染对照组的

且未见 GFAP表达。结论:mNotch-lshRNA抑制Notch-1基因的表达

促进MSCs向神经细胞的分化效率

提示MSCs的横向分化过程可能与神经干细胞类似

阻断Notch信号通路

会加MSCs向神经细胞分化的效果。

Abstract

Objectives; To identify the roles of Notch - 1 gene for survivaland differentiation of mesenchymal stem cells ( MSCs) into neurons by way of RNA interference ( RNAi) technique to suppress expression of Notch - 1 gene. Methods; Firstly

construct mNotch - 1 short hairpin RNA (mNotch -IshRNA) and then to oberve configuration of MSCs transfected by mNotch - IshRNA; MTT was employed to evaluate the survival ratio of MSCs before transfection

24 hours and 3 days after transfection

respectively; agents

such as BDNF

retinoid acid

were obtained to induce MSCs transfected by mNotch - IshRNA in vitro and expression of Nestin

NSE and NF200( neuron marker agent) and GFAP( neural glial cell marker agent) were determined by immunocytochemistry stain technique. Results: The cell cubic configuration was enforced and expression of notch - 1 gene vanished by transfection of mNotch -IshRNA. The survival ratio of MSCs by transfection decreased and there was significant differences between non - ransfection and transfection - controlling group after 24 hours and 3 days of transfection ( P <0. 01). The differentiation efficacy of MSCs transfected by mNotch - IshRNA into neurons and the expression of NSE

NF200 were increased significantly than non - ransfection and transfection -controlling group

and there was no expression of GFAP. Conclusions; The differentiation efficacy of MSCs can be enforced by mNotch - IshRNA to suppress Notch - 1 expression

which implies that the transverse differentiation of MSCs is similar to that of neural stem cells. The blocking of Notch - 1 signal pathway might promote differentiation of MSCs into neural stem cells.

关键词

骨髓间质干细胞Notch-1基因神经细胞shRNA

Keywords

Mesenchymal stem cells (MSCs)Notch-1 geneNeuronshRNA

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