首页  期刊介绍 征稿简则 编委会 期刊征订 广告服务 留言板 联系我们 English

中国应用生理学杂志 ›› 2019, Vol. 35 ›› Issue (4): 317-321.doi: 10.12047/j.cjap.5773.2019.067

• 研究论文 • 上一篇    下一篇

不同氧浓度下C2C12细胞的Nrf2抗氧化系统对H2O2刺激的反应*

姬卫秀1,2, 张缨1△   

  1. 1.北京体育大学, 北京 100084;
    2.天津体育学院, 天津 301617
  • 收稿日期:2018-10-24 出版日期:2019-07-28 发布日期:2019-11-06
  • 通讯作者: ,Tel: (010)62989584; E-mail: zhyi9256@126.com
  • 基金资助:
    * 国家自然科学基金项目(31471134)

The antioxidant system mediated by Nrf2 in C2C12 cells responding to H2O2 stimulus under different oxygen concentration

JI Wei-xiu1,2, ZHANG Ying1△   

  1. 1. Beijing Sport University, Beijing 100084;
    2. Tianjin University of Sport, Tianjin 301617, China
  • Received:2018-10-24 Online:2019-07-28 Published:2019-11-06

摘要: 目的:探讨不同氧浓度下小鼠骨骼肌卫星细胞系(C2C12细胞)对H2O2刺激反应的变化及其机制。方法:小鼠骨骼肌卫星细胞系(C2C12细胞),经培养复苏后,将细胞分为7组,每组设8个复孔,各组分别加入浓度为0.1 mmol/L、0.25 mmol/L、0.5 mmol/L、0.75 mmol/L、1 mmol/L、2 mmol/L的H2O2,分别作用1 h、2 h后测细胞活力,选择细胞H2O2刺激的最佳作用时间和浓度;C2C12细胞分为不同氧浓度组:21% O2、12% O2、8% O2、5% O2每组设8个复孔,12 h后,H2O2作用1 h,收集细胞;检测细胞Nrf2蛋白荧光和蛋白表达量,测定Nrf2和抗氧化酶SOD1、SOD2、CAT、NQO-1、HO-1、GPX-1 的mRNA表达量及细胞ROS水平。结果:选择H2O2作用时间相对较短的1 h和浓度0.5 mmol/L作为本实验的H2O2刺激条件。与21%O2组相比,12%O2组细胞Nrf2蛋白荧光增强,Nrf2 的mRNA和蛋白表达以及抗氧化酶SOD1、SOD2、CAT、NQO-1、HO-1、GPX-1的 mRNA表达均显著增加(P<0.05或P<0.01),细胞 ROS水平明显降低(P<0.01);8%O2组仅GPX-1 mRNA显著增加(P<0.05),其他指标变化不大;5%O2组细胞 Nrf2 mRNA和蛋白表达以及抗氧化酶SOD1、SOD2、NQO-1、GPX-1的 mRNA表达均明显降低(P<0.05或P<0.01),细胞 ROS水平则明显升高(P<0.01)。结论:不同氧浓度下C2C12细胞中Nrf2介导的抗氧化系统对H2O2刺激反应不同,12 h的12% O2浓度可促进C2C12细胞Nrf2的抗氧化作用,而5% O2浓度的严重低氧则作用相反。

关键词: 氧浓度, 过氧化氢, 小鼠骨骼肌卫星细胞系, 转录因子NF-E2相关因子, 抗氧化

Abstract: Objective: To apply hypoxia of different oxygen concentration on C2C12 cells to study the changes of Nrf2 antioxidant system under H2O2. Methods: The perfect simulative effect time and concentration of H2O2 were chosen. Cell vitality was tested after C2C12 cells cultured in 0.1 mmol/L, 0.25 mmol/L, 0.5 mmol/L, 0.75 mmol/L, 1 mmol/L and 2 mmol/L H2O2 for 1 or 2 h respectively. The C2C12 cells were divided into different oxygen concentration group: 21%O2, 12%O2, 8%O2, 5%O2 respectively. And then cells were treated with H2O2 for 1 h, and collected for determination. Immunofluorescence of Nrf2 and the protein expression of Nrf2 were detected. The expressions of antioxidant enzymes superoxide dismutase 1 (SOD1), superoxide dismutase 2 (SOD2), catalase(CAT), NADPH quinine oxidoreductase-1 (NQO-1), glutathione peroxidase-1 (GPX-1), Heme oxygenase-1 (HO-1) mRNA and cellular ROS levels were tested by high quality fluorescence assay. Results: 0.5 mmol/L H2O2 for 1 h was selected as the conditions of H2O2stimulation. Compared with 21% O2 group, the expressions of Nrf2 mRNA and protein, antioxidant enzymes SOD1, SOD2, CAT, HO-1, NQO-1, GPX-1 mRNA were increased significantly (P<0.05 or P<0.01), and ROS level was lower (P<0.01) in 12%O2 group cells; only the expression of GPX-1 mRNA was increased (P<0.05) in 8%O2 group; the expressions of Nrf2 mRNA and protein expression, antioxidant enzymes SOD1, SOD2, NQO-1, GPX-1 mRNA were decreased significantly(P<0.05 or P<0.01), and ROS level was higher (P<0.01) in 5%O2 group. Conclusion: Hypoxia can affect the Nrf2 antioxidant system, and the different oxygen concentrations have different impact. In addition, 12% O2 for 12 h could promote the Nrf2 antioxidant system, and 5% extremely low oxygen may inhibit it.

Key words: oxygen concentration, H2O2, C2C12 cells, Nrf2, antioxidant

版权所有 © 2015 《中国应用生理学杂志》编辑部
京ICP备16058274号-1
地址:天津市和平区大理道1号,邮编:300050  电话:022-84655184  E-mail:tjzgyish@163.com
本系统由北京玛格泰克科技发展有限公司设计开发 技术支持:support@magtech.com.cn