中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (15): 2751-2754.doi: 10.3969/j.issn.1673-8225.2011.15.022

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

p38MAPK信号通路与应力介导成肌细胞的凋亡

田  臻1，杨竹丽2，贾文敏3，袁  晓2，仇  静1，达  雨1，杜衍晓1，于江波2，张  月2，刘  文2   

  1. 1 青岛大学医学院附属青岛市市立医院口腔医学中心，山东省青岛市  266071
    2 青岛市市立医院口腔科，山东省青岛市  266071
    3 解放军济南军区青岛第一疗养院口腔科，山东省青岛市  266021
  • 收稿日期:2010-12-15 修回日期:2011-02-16 出版日期:2011-04-09 发布日期:2013-11-06
  • 通讯作者: 袁晓，博士，副主任医师，青岛市市立医院口腔科，山东省青岛市 266071
  • 作者简介:田臻★，女，1984年生，山东省青岛市人，汉族，青岛大学医学院在读硕士，主要从事口腔正畸学研究。
  • 基金资助:

    国家自然科学基金资助项目(30871426)。

Role of p38MAPK signaling pathways in the apoptosis of C2C12 myoblast cells subjected to cyclical stretch

Tian Zhen1, Yang Zhu-li2, Jia Wen-min3, Yuan Xiao2, Qiu Jing1, Da Yu1, Du Yan-xiao1, Yu Jiang-bo2, Zhang Yue2, Liu Wen2   

  1. 1Stomatology Center of Qingdao Municipal Hospital, Qingdao University Medical College, Qingdao  266071, Shandong Province, China
    2Department of Stomatology, Qingdao Municipal Hospital, Qingdao  266071, Shandong Province, China
    3Department of Stomatology, Qingdao 1st Sanatorium of Jinan Military Command, Qingdao  266021, Shandong Province, China 
  • Received:2010-12-15 Revised:2011-02-16 Online:2011-04-09 Published:2013-11-06
  • Contact: Yuan Xiao, Doctor, Associate chief physician, Department of Stomatology, Qingdao Municipal Hospital, Qingdao 266071, Shandong Province, China yuanxiaoqd@163. com
  • About author:Tian Zhen★, Studying for master’s degree, Stomatology Center of Qingdao Municipal Hospital, Qingdao University Medical College, Qingdao 266071, Shandong Province, China tianzhen_0072@163.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30871426*

PDF

436

被引次数

13

摘要：

背景：在体内条件下，细胞力学的功能研究因其所处生理环境的复杂性、实验条件的不易控制而很难得到满意结果。
目的：在成功构建成肌细胞体外培养-力学刺激模型的基础上，研究p38MAPK信号通路在成肌细胞凋亡中的作用及其机制。
方法：将体外培养的C2C12细胞分为对照组和SB203580组，SB203580组中加入  20 mmol/L的p38MAPK抑制剂SB203580。应用细胞应力加载装置Flecell Strain Unit-5000T给细胞提供15%的力值，分别施加0，6，12，24 h的周期性张应力。每分钟10个循环，每循环包括3 s牵张，3 s松弛。Hoechst 33258染色观察细胞的形态学变化；流式细胞仪检测细胞凋亡情况；RT-PCR法检测促凋亡基因bax mRNA的表达；Western blot检测信号通路中p38MAPK和p-p38MAPK蛋白的表达。
结果与结论：随着加力时间的延长，细胞逐渐出现核固缩及凋亡小体，凋亡率增加(P < 0.05)，bax mRNA表达增多(P < 0.05)；细胞p38MAPK和p-p38MAPK蛋白均在加力6 h达到最低，此后逐渐升高。p38MAPK抑制剂SB203580可抑制加力引起的细胞凋亡，减少bax mRNA及p38MAPK和p-p38MAPK蛋白的表达(P < 0.05)。说明p38MAPK信号通路在应力介导的成肌细胞凋亡中起到重要的作用。

关键词: 细胞凋亡, C2C12, p38MAPK, 周期性张应力, 成肌细胞

Abstract:

BACKGROUND: Because of complicated physiological environment and difficulty to control experimental conditions, it is difficult to get satisfactory results from in vivo studies of cell mechanics.
OBJECTIVE: To study the action and mechanism of p38MAPK signaling pathways on myoblast apoptosis based on successful construction of in vitro mechanical stimulation models.
METHODS: The C2C12 cells cultured in vitro were divided into control group and SB203580 treatment group. Cyclic tensile stress was applied on the C2C12 myoblast cells for 0, 6, 12 and 24 hours in each group. The Flexcell Strain Unit-5000T was used to expose C2C12 myoblast cell to an equiaxial cyclic of 15% magnitude and a frequency of 10 cycles/min, each cycle including the 3 s stretch and 3 s relaxation. Hoechst 33258 fluorescent staining and optical microscope were used to detect cell apoptosis. RT-PCR, flow cytometric analysis were used to observe the apoptosis of C2C12 myoblast cells and Western blotting were used to detect the activity of p38MAPK and p-p38MAPK.
RESULTS AND CONCLUSION: The optical microscope tested the change in the morphology. Hoechst 33258 staining showed that after treatment with cyclic stress, the cell took the typical appearance of apoptosis with chromatin condensation and apoptotic bodies. RT-PCR and flow cytometry showed that with the extension of time the rate of the apoptosis of C2C12 myoblast cell increased. And cells imposed SB203580 before imposing cyclical tensile stress, the results showed that the apoptosis was markedly affected, and the p-p38MAPK expression declined apparently. These findings demonstrate that p38MAPK signaling pathways in stress mediated into C2C12 myoblast cell apoptosis plays an important role.

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