Notch信号通路在骨髓间充质干细胞向肝细胞分化过程中的动态表达特征

中国临床解剖学杂志 ›› 2013, Vol. 31 ›› Issue (3): 308-313.

Notch信号通路在骨髓间充质干细胞向肝细胞分化过程中的动态表达特征

柳柯，　刘桂英，　杨洋，　吴章林，　黄文华

南方医科大学人体解剖学教研室广东省组织构建与检测重点实验室，广州 510515

收稿日期:2013-01-28 出版日期:2013-05-25 发布日期:2013-06-06
通讯作者: 黄文华，博士生导师，教授，E-mail：huangwenhua2009@　139.com E-mail:liuke20050@yahoo.com.cn
作者简介:柳柯（1987-），男，江西九江人，在读硕士，研究方向：主要丛事组织工程方向研究，Tel：13697411099
基金资助:
国家科技部863项目分题（2012AA02A603）

Dynamic expression characteristics of Notch signal in bone marrow-derived mesenchymal stem cells during the process of differentiation into hepatocytes

LIU Ke，LIU Gui-ying, YANG Yang, WU Zhang-lin, HUANG Wen-hua

Department of Anatomy,Southern Medical University，Guangzhou 510515，China

Received:2013-01-28 Online:2013-05-25 Published:2013-06-06

摘要/Abstract

摘要：

目的　探寻Notch信号通路对BM-MSCs向肝细胞分化的影响机制。方法　诱导BM-MSCs分化成肝细胞。当BM-MSCs分化至第0、7、11、21天时，反向斑点杂交实验检测Notch信号通路中的关键基因的mRNA表达水平。建立加入Jagged1上调信号通路的对照组RT-PCR技术绘制出BM-MSCs分化状态分子表达谱与正常情况下对比。结果　BM-MSCs分化进行至第21天，反向斑点杂交检测到的关键基因的mRNA表达水平低于第0、7、11天。加入Jagged1后Notch信号通路被激活，导致下游基因Hes1 和 Hey1的被表达。BM-MSCs分化过程中Albumin未被检测到。结论　本研究的结果表明Notch信号通路在BMMSCs分化成肝细胞过程进行调控是必需的，但是分化必须在信号通路下调的情况下才能进行下去。

关键词: 成体干细胞, 间充在干细胞, 定向分化, 肝细胞, Notch信号通路

Abstract:

Objective　To explore the effect of Notch signaling in bone marrow-derived mesenchymal stem cells during the process of differentiation into hepatocytes.　Methods　An induction system under which BM-MSCs can differentiate into hepatocytes was established. On days 0, 7, 11 and 21 day during the differentiation direction, nine key genes in Notch pathway were selected as target genes and determined in BM-MSCs using technique reverse dot blot hybridization. The results of reverse dot blot hybridization showed that the effect of Notch signaling in BM-MSCs during the process of differentiation into hepatocytes. At the same time，the results of RT-PCR analysis reveals the differentiation status of BM-MSCs. Results On the 21th day when the differentiation direction was determined in BM-MSCs, the mRNA level of nine key genes was significantly lower than that on days 0, 7, and 11. In the further experiments, down-regulation of Notch signaling was shown to be critical for BM-MSCs to differentiate into hepatocytes, as increased Jagged1 resulted in up-regulated Notch activation, leading to higher levels of expression of Hes1 and Hey1, which completely blocked Albumin expresion in BM-MSCs. Conclusion These results in our study showed that Notch signaling in BM-MSCs was necessary to initiate differentiation into hepatocytes, but must be down-regulated for the differentiation to proceed continuously.

Key words: Adult stem cell, Mesenchymal stem cell, Directing-differentiation, Hepatocyte, Notch

中图分类号:

R329.2

柳柯, 刘桂英, 杨洋, 吴章林, 黄文华. Notch信号通路在骨髓间充质干细胞向肝细胞分化过程中的动态表达特征[J]. 中国临床解剖学杂志, 2013, 31(3): 308-313.

LIU Ke, LIU Gui-Yang, YANG Xiang, TUN Zhang-Lin, HUANG Wen-Hua. Dynamic expression characteristics of Notch signal in bone marrow-derived mesenchymal stem cells during the process of differentiation into hepatocytes[J]. Chinese Journal Of Clinical Anatomy, 2013, 31(3): 308-313.

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