华西口腔医学杂志

• 基础研究 • 上一篇    下一篇

钙调神经磷酸酶-T细胞核因子信号通路在应力诱导成肌细胞凋亡中的作用

丁弦1 夏晨蕾1 贺苗1 孙文娜1 王芳1 姜文心1 张彩霞1 王爽玉1 张强1 姚如永2 袁晓1   

  1. 1.青岛大学医学院附属青岛市立医院口腔医学中心,青岛 266075;2.青岛大学医学院附属医院中心实验室,青岛 266003
  • 收稿日期:2015-04-15 修回日期:2015-07-20 出版日期:2015-10-01 发布日期:2015-10-01
  • 通讯作者: 袁晓,教授,博士,E-mail:yuanxiaoqd@163.com
  • 作者简介:丁弦,硕士,E-mail:xiaowunvdx@163.com
  • 基金资助:

    国家自然科学基金资助项目(31170891);山东省卫生厅重点基金资助项目(2011HD001)

Role of calcineurin-nuclear factor of activated T cells signaling pathway in myoblast apoptosis induced by cyclic tensile strain

Ding Xian1, Xia Chenlei1, He Miao1, Sun Wenna1, Wang Fang1, Jiang Wenxin1, Zhang Caixia1, Wang Shuangyu1, Zhang Qiang1, Yao Ruyong2, Yuan Xiao1.   

  1. 1. Dept. of Stomatology, The Affiliated Qingdao Municipal Hospital, Medical College of Qingdao University, Qingdao 266075, China; 2. The Central Laboratory, The Affiliated Hospital of Medical College of Qingdao University, Qingdao 266003, China
  • Received:2015-04-15 Revised:2015-07-20 Online:2015-10-01 Published:2015-10-01

摘要:

目的  探讨钙调神经磷酸酶(CaN)-T细胞核因子(NFAT)信号通路在应力介导的成肌细胞凋亡中的作用。方法  构建成肌细胞体外培养—力学刺激模型,利用多通道应力加载系统对细胞加载不同时间的周期性张应力,加入CaN的特异性抑制剂环孢素(CsA)作为对比。采用Hoechst 33258染色法和流式细胞术检测成肌细胞凋亡情况,实时聚合酶链式反应检测CaN和NFAT mRNA的表达情况,蛋白质印迹法检测NFAT3的蛋白含量。结果  随加力时间的延长,细胞凋亡逐渐增加,CaN亚基CnA、CnB及NFAT3的mRNA表达及NFAT3蛋白含量逐渐升高;加入 CsA后,细胞凋亡减少,CnA、NFAT3的mRNA表达及NFAT3的蛋白含量明显减少。结论  周期性张应力可以诱导成肌细胞发生凋亡;CaN-NFAT信号通路可能参与了周期性张应力诱导的成肌细胞凋亡。

关键词: 成肌细胞, 细胞凋亡, 钙调神经磷酸酶, T细胞核因子, 周期性张应力

Abstract:

Objective  This study investigated the role and mechanism of calcineurin (CaN)-nuclear factor of activated T cells (NFAT) pathway in the myoblast apoptosis induced by cyclic tensile strain. Methods  Myoblasts were cultured using an in vitro-mechanical stimulation model and imposed with tension for different hours with a multi-channel cell stress loading system. Cyclosporine (CsA) was used as CaN inhibitor to clarify the role of CaN in the apoptosis induced by cyclic stress. Hochest 33258 staining and flow cytometry detection were performed to detect the apoptotic cells. Real-time polymerase chain reaction was conducted to detect the mRNA expression of CaN and NFAT. Protein levels of NFAT3 were evaluated by Western blot. Results  The apoptosis rate increased with the extension of loading time. The mRNA expression of the CaN subunits, CnA and CnB, and the protein levels of NFAT3 also increased. When the myoblasts were incubated with CsA, the apoptosis rate decreased, the mRNA expression of CnA and NFAT3 significantly decreased, and the NFAT3 protein expression levels became significantly lower than those of the groups without CsA. Conclusion  Continuous cyclic tensile stress can induce myoblast apoptosis. The CaN-NFAT signaling pathway may be involved in the cyclic stretch-induced apoptosis of myoblasts.

Key words: myoblasts, apoptosis, calcineurin, nuclear factor of activated T cells, cyclic tensile strain