中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (23): 4193-4198.doi: 10.3969/j.issn.1673-8225.2012.23.003

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

小鼠骨髓间充质干细胞体外成肌分化中基质金属蛋白酶1的作用

郑振扬1，张为西1，冷 雁2，周 琛1，马震宇1，钟志刚1   

  1. 中山大学附属第一医院，1神经科，2康复科，广东省广州市 510080
  • 收稿日期:2011-11-08 修回日期:2012-01-07 出版日期:2012-06-03 发布日期:2013-11-06
  • 通讯作者: 张为西，博士，教授，主任医师，博士生导师，中山大学附属第一医院神经科，广东省广州市 510080 weixizhang@qq.com
  • 作者简介:郑振扬★，男，1986年生，福建省莆田市人，汉族，中山大学附属第一医院神经科在读硕士，主要从事骨髓间充质干细胞与肌肉疾病的研究。 xiaotang_1986@yahoo.com.cn
  • 基金资助:

    国家自然科学基金资助项目(30971026，30870852)

Effect of matrix metalloproteinase-1 on myogenic differentiation of mouse bone marrow-derived mesenchymal stem cells in vitro

Zheng Zhen-yang1, Zhang Wei-xi1, Leng Yan2, Zhou Chen1, Ma Zhen-yu1, Zhong Zhi-gang1   

  1. 1Department of Neurology, 2Department of Rehabilitation, First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080, Guangdong Province, China
  • Received:2011-11-08 Revised:2012-01-07 Online:2012-06-03 Published:2013-11-06
  • Contact: Zhang Wei-xi, Doctor, Professor, Chief physician, Doctor’s supervisor, Department of Neurology, First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080, Guangdong Province, China weixizhang@qq.com.
  • About author:Zheng Zhen-yang★, Studying for master’s degree, Department of Neurology, First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080, Guangdong Province, China xiaotang_1986@yahoo.com.cn.

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摘要：

背景：小鼠骨髓间充质干细胞体外成肌诱导分化效率较低。
目的：观察基质金属蛋白酶1在体外对小鼠骨髓间充质干细胞成肌分化的作用。
方法：利用差速贴壁法体外分离培养小鼠骨髓间充质干细胞并进行鉴定后，按照不同基质金属蛋白酶1药物处理浓度将小鼠骨髓间充质干细胞分成4组，即10 μg/L组、1 μg/L组、0.1 μg/L组、对照组，给予相应处理后，Real-time定量PCR检测MyoD、Desmin mRNA表达，Western Blotting检测Desmin蛋白表达。
结果与结论：利用差速贴壁法分离培养的小鼠骨髓间充质干细胞形态较一致，表面分子检测示CD29+、Sca-1+、CD45-、CD34-，并具有多向分化潜能；经基质金属蛋白酶1处理后，Real-time定量PCR检测示Desmin、MyoD mRNA表达上调，Western Blotting检测示Desmin蛋白表达上调，并且以上各成肌相关基因、蛋白表达增高程度与基质金属蛋白酶1具有浓度依赖性。提示基质金属蛋白酶1在体外可促进骨髓间充质干细胞向肌肉细胞分化。

关键词: 基质金属蛋白酶1, 骨髓间充质干细胞, 成肌分化, 5-氮胞苷, 小鼠, 干细胞

Abstract:

BACKGROUND: The myogenic differentiation potential of bone marrow-derived mesenchymal stem cells (MSCs) in vitro from mice is low.
OBJECTIVE: To explore the effect of matrix metalloproteinase-1 (MMP-1) on myogenic differentiation of MSCs in vitro from mice.
METHODS: After MSCs were isolated and cultured by differential adhesion, as well as identified by cell surface markers and multi-lineage differentiation potential, the cells were assigned to 4 groups according to various MMP-1 concentration gradients (10 μg/L group, 1 μg/L group, 0.1 μg/L group and control group). The mRNA expressions of MyoD and Desmin were detected by real-time quantitative PCR, and the expression of protein Desmin was tested by Western Blotting.
RESULTS AND CONCLUSION: MSCs obtained from differential adhesion were of morphological homogenicity, and they were CD29+, Sca-1+, CD45- and CD34-. Additionally, MSCs were capable of multi-lineage differentiation. The mRNA expression of Desmin and MyoD as well as the expression of protein Desmin were up regulated via MMP-1 inducement. Moreover, the up-regulation degree of mRNA and protein expression was concentration-dependent. These suggest that MMP-1 can promote MSCs to differentiate into muscle cells in vitro.

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