Chinese Medical Sciences Journal ›› 2018, Vol. 33 ›› Issue (1): 45-52.doi: 10.24920/31801

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  1. 西安交通大学医学部公共卫生学院 地方病研究所, 西安 710061
    西安交通大学医学部研究生教学实验中心,西安 710061
  • 收稿日期:2017-02-20 出版日期:2018-03-07 发布日期:2018-03-07
  • 通讯作者: 陈静

Gene Expression Profile of Hypertrophic Chondrocytes Treated with H2O2: A Preliminary Investigation

He Ying1,2,Zhang Ying1,Wang Mengying1,Zhang Meng1,Zhang Dan1,Zhang Ying1,Jiang Zhuocheng1,Wu Feng2,Chen Jinghong1,*   

  1. 1 Institute of Endemic Diseases, School of Public Health, Xi’an Jiaotong University College of Medicine, Xi’an 710061, China
    2 Graduate Students Teaching Experiment Center, Xi’an Jiaotong University College of Medicine, Xi’an 710061, China
  • Received:2017-02-20 Published:2018-03-07 Online:2018-03-07
  • Contact: Chen Jinghong

摘要: 目的 筛选双氧水可以引起哪些骨生成相关基因的改变。方法 用胰岛素-转铁蛋白和亚硒酸钠的混合液诱导鼠软骨前体细胞分化为肥大软骨细胞。采用MTT法确定双氧水作用的最佳浓度和时间。采用PCR阵列检测细胞中84个骨生成相关基因的表达;并用定量RT-PCR验证PCR 阵列结果。结果 结果显示:9个基因表达上调、12个基因表达下调,这些基因编码多种功能蛋白,其中包括胶原蛋白、转录因子、骨骼发育和骨矿物质代谢相关蛋白以及细胞粘附分子等。定量RT-PCR验证了5个表达下调基因(Smad2,Smad4,转化生长因子βr1、βr3,以及基质金属蛋白酶10)。结论 双氧水改变了一些具有不同生物学功能的基因在肥大软骨细胞中的表达。结合氧化损伤与Kashin-Beck病相关的文献,我们推测这些基因有可能参与了Kashin-Beck病软骨的深层坏死。

关键词: Kashin-Beck病, 肥大软骨细胞, 深层坏死, 氧化应激, 双氧水

Abstract: Objective To identify the osteogenesis genes whose expression is altered in hypertrophic chondrocytes treated with H2O2.Methods Murine chondrogenitor cells (ATDC5) were differentiated into hypertrophic chondrocytes by Insulin-Transferrin-Selenium (ITS) treatment, and then treated with H2O2. Suitable conditions (concentration, time) were determined by using the MTT assay. After total RNA isolation and cDNA synthesis, the levels of 84 genes were determined using the PCR array, whereas quantitative RT-PCR was carried out to validate the PCR array data.Results We identified 9 up-regulated genes and 12 down-regulated genes, encoding proteins with various functions, such as collagen proteins, transcription factors, proteins involved in skeletal development and bone mineral metabolism, as well as cell adhesion molecules. Quantitative RT-PCR confirmed the altered expression of 5 down-regulated genes (Smad2, Smad4, transforming growth factor β receptor 1, transforming growth factor β receptor 3, and matrix metalloproteinase 10).Conclusions H2O2 significantly changed the expression of several genes involved in a variety of biological functions. Because of the link between oxidative damage and Kashin-Beck disease, these genes may also be involved in the deep-zone necrosis of the cartilage observed in Kashin-Beck disease.

Key words: Kashin-Beck disease, hypertrophic chondrocytes, deep-zone necrosis, oxidative stress, H2O2

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