中国组织工程研究

中国组织工程研究 ›› 2014, Vol. 18 ›› Issue (20): 3236-3241.doi: 10.3969/j.issn.2095-4344.2014.20.022

• 骨组织构建 bone tissue construction • 上一篇    下一篇

骨形态发生蛋白2诱导C2C12和MC3T3-E1的成骨分化与自噬

高  艳，程  晨，李  静，张  越，肖伟凡，潘秋辉   

  1. 同济大学附属第十人民医院中心实验室，上海市  200072
  • 收稿日期:2014-02-15 出版日期:2014-05-14 发布日期:2014-05-14
  • 通讯作者: 潘秋辉，博士，教授，同济大学附属第十人民医院中心实验室，上海市 200072
  • 作者简介:高艳，女，1986年生，山东省日照市人，汉族，同济大学在读硕士，主要从事骨分化发育的研究。
  • 基金资助:

    国家自然科学基金项目(81171178)

Bone morphogenetic protein 2-induced C2C12 and MC3T3-E1 osteoblast differentiation and autophagy

Gao Yan, Cheng Chen, Li Jing, Zhang Yue, Xiao Wei-fan, Pan Qiu-hui   

  1. Central Laboratory, Tenth People’s Hospital, Tongji University, Shanghai 200072, China
  • Received:2014-02-15 Online:2014-05-14 Published:2014-05-14
  • Contact: Pan Qiu-hui, M.D., Professor, Central Laboratory, Tenth People’s Hospital, Tongji University, Shanghai 200072, China
  • About author:Gao Yan, Studying for master’s degree, Central Laboratory, Tenth People’s Hospital, Tongji University, Shanghai 200072, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81171178

PDF

648

被引次数

2

摘要：

背景：一系列研究表明自噬与分化有密切联系；骨形态发生蛋白2是诱导C2C12、MC3T3-E1成骨分化经典途径，是研究成骨分化过程的理想模型。
目的：观察自噬与骨形态发生蛋白2诱导细胞株C2C12、MC3T3-E1成骨分化的关系。
方法：Real-Time PCR检测MC3T3-E1与C2C12在骨形态发生蛋白2(100 μg/L)诱导培养3 d后成骨与自噬相关指标变化。碱性磷酸酶染色检测不同浓度3-甲基腺嘌呤(0，1，5，10 mmol/L)对骨形态发生蛋白2(100 μg/L)诱导培养7 d MC3T3-E1与C2C12成骨指标碱性磷酸酶变化，Western Blot检测C2C12和MC3T3-E1在骨形态发生蛋白2(100 μg/L)诱导不同时间点(0，12，24，48，72，96 h)LC3-Ⅰ/Ⅱ蛋白表达水平。
结果与结论：在骨形态发生蛋白2诱导细胞株C2C12、MC3T3-E1成骨分化过程中，诱导自噬相关mRNA与蛋白水平均有增高趋势，且自噬相关蛋白LC3水平增高与时间相关。同时，抑制自噬成骨分化过程中碱性磷酸酶表达水平降低。因此，自噬与骨形态发生蛋白2诱导细胞株C2C12、MC3T3-E1成骨分化过程有密切关系。



中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程


全文链接：

关键词: 组织构建, 骨组织工程, 自噬, 骨形态发生蛋白2, C2C12, MC3T3-E1, 成骨分化, 国家自然科学基金

Abstract:

BACKGROUND: A series of studies indicate that autophagy is closely linked with differentiation. Bone morphogenetic protein 2 (BMP-2) is the classical pathway for C2C12 and MC3T3-E1 osteoblast differentiation, and the ideal model to study osteogenic differentiation process.
OBJECTIVE: To observe the relationship between autophagy and BMP-2-induced cell lines C2C12, MC3T3-E1 osteoblast differentiation.
METHODS: Real-Time PCR was applied to detect osteogenic differentiation and autophagy related index after C2C12 and MC3T3-E1 were induced with BMP-2 (100 μg/L) for 72 hours. The osteogenic index alkaline phosphatase in BMP-2-induced MC3T3-E1 and C2C12 cultured with different concentrations of 3-methyladenine (0, 1, 5, 10 mmol/L) was determined with alkaline phosphatase staining. Western blot analysis was applied to detect LC3-I/II expression levels in C2C12 and MC3T3-E1 induced with BMP-2 for different time points (0, 12, 24, 48, 72, 96 hours).
RESULTS AND CONCLUSION: The autophagy-related mRNA and protein expression showed an increasing tendency and autophagy-related protein LC3 levels was increased, which was associated with the time, during the BMP-2-induced cell lines C2C12, MC3T3-E1 osteoblast differentiation. Meanwhile, alkaline phosphatase expression levels were inhibited by autophagy in the process of osteogenic differentiation. Therefore, there is a close relationship between autophagy and the BMP-2-induced cell lines C2C12, MC3T3-E1 osteoblast differentiation.



中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程


全文链接：

Key words: autophagy, bone morphogenetic proteins, cell line, myoblasts, osteoblasts

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