中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (32): 5920-5925.doi: 10.3969/j.issn.2095-4344.2012.32.006

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

胡 波1，高 攀2，马志峰1，张志峰3，张中明1，袁红花3，董红燕3   

  1. 1徐州医学院附属医院胸心外科，江苏省徐州市 221006；
    2商丘市第一人民医院心血管外科，河南省商丘市 476100；
    3徐州医学院神经生物学研究中心实验室/生物学教研室，江苏省徐州市 221006
  • 收稿日期:2011-12-20 修回日期:2012-02-15 出版日期:2012-08-05 发布日期:2012-08-05
  • 通讯作者: 董红燕，教授，徐州医学院神经生物学研究中心实验室/生物学教研室，江苏省徐州市 221006
  • 作者简介:胡波★，男，1972年生，安徽省合肥市人，汉族，2003年徐州医学院毕业，硕士，副主任医师，主要从事骨髓间充质干细胞向心肌样细胞分化及其机制的研究。

Bone marrow-derived mesenchymal stem cells differentiation into cardiomyocyte-like cells in myocardial microenvironment

Hu Bo1, Gao Pan2, Ma Zhi-feng1, Zhang Zhi-feng3, Zhang Zhong-ming1, Yuan Hong-hua3, Dong Hong-yan3   

  1. 1Department of Cardio-Thoracic Surgery, Xuzhou Medical College, Xuzhou 221006, Jiangsu Province, China;
    2Department of Cardiovascular Surgery, Shangqiu First People’s Hospital, Shangqiu 476100, Henan Province, China;
    3Central Laboratory of Neurobiology/ Biology Teaching and Research Section, Xuzhou Medical College, Xuzhou 221006, Jiangsu Province, China
  • Received:2011-12-20 Revised:2012-02-15 Online:2012-08-05 Published:2012-08-05
  • Contact: Dong Hong-yan, Professor, Central Laboratory of Neurobiology/Biology Teaching and Research Section, Xuzhou Medical College, Xuzhou 221006, Jiangsu Province, China dhyxzh@yahoo.com.cn
  • About author:Hu Bo★, Master, Associate chief physician, Department of Cardio-Thoracic Surgery, Xuzhou Medical College, Xuzhou 221006, Jiangsu Province, China Feeling6@163.com

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摘要：

背景：骨形态发生蛋白、Wnt、Notch、FGF及Hedgehog等信号通路是否参与诱导骨髓间充质干细胞向心肌细胞的分化还不清楚。
目的：探讨心肌微环境中，不同刺激因素诱导骨髓间充质干细胞分化为心肌样细胞及其分化的机制。
方法：利用Transwell培养装置建立心肌微环境，骨髓间充质干细胞在此环境中培养并加入Wnt11、骨形态发生蛋白抑制剂诱导其向心肌细胞分化；应用RT-PCR、Western blot方法检测骨形态发生蛋白、Wnt、Notch、FGF和Hedgehog信号通路关键信号分子的表达。同时检测心肌特异性转录因子(Nkx2.5、GATA4、Mef2c)和成熟心肌细胞特异性基因(cTnI、ANP、α-MHC、β-MHC)的表达。
结果与结论：骨髓间充质干细胞经过与大鼠心肌细胞共培养诱导后，Wnt、骨形态发生蛋白、Notch、Hedgehog信号通路关键信号分子的表达较未诱导的阴性对照组明显升高(P < 0.01)。在骨髓间充质干细胞与大鼠心肌细胞共培养体系中加入骨形态发生蛋白信号通路的抑制剂Noggin，心肌特异性转录因子或结构蛋白的表达明显降低(P < 0.01)。提示心肌细胞构成的微环境可以诱导骨髓间充质干细胞分化为心肌样细胞；Wnt、骨形态发生蛋白、Notch、FGF、Hedgehog信号通路共同参与了诱导分化过程。

关键词: 心肌微环境, 骨髓间充质干细胞, Wnt11, 骨形态发生蛋白2, 诱导分化, 心肌样细胞

Abstract:

BACKGROUND: It remains unclear whether bone morphogenetic protein (BMP), Wnt, Notch, FGF and Hedgehog signaling pathways are involved in differentiation of cardiomyocytes from bone marrow-derived mesenchymal stem cells (BMSCs).
OBJECTIVE: To investigate the ability of BMSCs differentiating into cardiomyocytes-like cells and their differentiating mechanism of signal pathways in myocardial microenvironment.
METHODS: Myocardial microenvironment was established by the technology of Transwell, and Wnt11 and BMP2 were used to induce BMSCs differentiation into cardiomyocyte-like cells. RT-PCR and western blot were used to detect expression of key signaling molecules of BMP, Wnt, Notch, FGF and Hedgehog signaling pathways. In addition, expression of cardiac-specific transcription factors, Nkx2.5, GATA4, and Mef2c, and mature cardiac-specific genes, cTnI, ANP, α-MHC, and β-MHC, was detected.
RESULTS AND CONCLUSION: After co-culture with rat cardiomyocytes, in the BMSCs, the expression of key signaling molecules of Wnt, BMP, Notch, Hedgehog signal pathways was significantly higher than the non-induced negative control group (P < 0.01). After adding BMP signal pathway inhibitor, Noggin, cardiac-specific transcription factors or structural proteins Nkx-2.5, GATA4, α-MHC, β-MHC, cTnI expression was significantly reduced (P < 0.01). Results indicate that myocardial microenvironment can induce rat BMSCs to differentiate into cardiomyocyte-like cells. Wnt, BMP, Notch, FGF and Hedgehog signaling pathways were all involved in the process.

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