山东大学耳鼻喉眼学报 ›› 2015, Vol. 29 ›› Issue (6): 56-59.doi: 10.6040/j.issn.1673-3770.0.2015.215

• 论著 • 上一篇    下一篇

SIRT1抗视网膜色素上皮细胞氧化应激作用的实验研究

李兰根1, 伟伟1, 张玉凤1, 格日乐图1, 杨佳2, 张艳梅3   

  1. 1. 内蒙古自治区人民医院眼科, 内蒙古 呼和浩特 010017;
    2. 内蒙古医科大学附属医院神经内科, 内蒙古 呼和浩特 010000;
    3. 内蒙古自治区人民医院神经内科, 内蒙古 呼和浩特 010017
  • 收稿日期:2015-06-02 出版日期:2015-12-16 发布日期:2015-12-16
  • 通讯作者: 张艳梅. E-mail:zhangyanmei2277@sina.com E-mail:zhangyanmei2277@sina.com
  • 作者简介:李兰根. E-mail:2807278584@qq.com

The experiment of SIRT1 on against oxidative stress to retinal pigmented epithelium cells

LI Langen1, WEI Wei1, ZHANG Yufeng1, Geriletu1, YANG Jia2, ZHANG Yanmei3   

  1. 1. Departments of Ophthalmology, Inner Mongolia People's Hospital, Hohhot 010017, Inner Mongolia, China;
    2. Departments of Neurology, Affiliated Hospital of Inner Mongolia Medical University, Hohhot 010000, Inner Mongolia, China;
    3. Departments of Neurology, Inner Mongolia People's Hospital, Hohhot 010017, Inner Mongolia, China
  • Received:2015-06-02 Online:2015-12-16 Published:2015-12-16

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摘要： 目的 观察去乙酰化酶-1(SIRT1)对视网膜色素上皮细胞(RPE)氧化应激的防护作用.方法 应用50 μmol/L H2O2与ARPE-19细胞孵育24 h诱导RPE产生氧化应激病理生理过程,观察细胞增殖及凋亡率、细胞活性氧水平(ROS), 氧化剂诱导RPE细胞衰老状况,了解氧化应激对RPE的损伤;进一步采用SIRT1激动剂白藜芦醇(RSV)及抑制剂烟酰胺(NA)分别处理经H2O2干预的RPE细胞,观察SIRT1抗RPE氧化应激效果.结果 ①与对照组相比,H2O2干预RPE后细胞增殖显著下降(P=0.020),凋亡率及内生ROS水平均显著增高(P=0.013, P=0.040),细胞衰老水平显著升高(P=0.045);②NA可显著提高H2O2的毒性作用,降低RPE细胞增殖率(P=0.049),刺激凋亡(P=0.028),使ROS累积和衰老速度增加;③RSV具有显著的抗H2O2毒性作用;④RT-qPCR分析显示,SIRT1mRNA水平显著下调.结论 氧化应激状况下SIRT1表达下调,表明SIRT1对氧化应激诱发的视网膜色素上皮细胞损伤有显著的防护作用.适当提高SIRT1活性也许是延缓AMD的一种治疗策略.

关键词: H2O2, 乙酰化酶-1, 视网膜色素上皮细胞, 细胞生长, 细胞活性氧水平, siRNA, 氧化应激

Abstract: Objective To observe the protection role of the sirtuin 1 (SIRT1) towards the retinal pigmented epithelium(RPE) cells following exposure to oxidative stress. Methods The rates of proliferation and apoptosis, levels of intracellular reactive oxygen species(ROS) and cell senescence of RPEs, induced by oxidants (H2O2), were evaluated. Results The results revealed a down-regulation of SIRT1 expression during oxidative stress. Furthermore, SIRT1 activator resveratrol (RSV) and inhibitors nicotinamide (NA) were respectively treated on RPE cells following exposure by H2O2 intervention; the expression of SIRT1 gene were detected by RT-PCR technology after RNA interfering on SIRT1. The experiments indicated that cell proliferation, apoptosis and ROS levels were significantly different compared with the control group after intervention by H2O2, and cell senescence were significantly increased; NA was significantly increased the toxicity of H2O2, however RSV had the opposite anti-cytotoxicity of H2O2; RT-PCR analysis showed that SIRT1 levels were significantly reduced. Conclusion These results therefore suggested that down-regulation of SirT1 expression during oxidative stress, further investigation indicated that SIRT1 protected RPEs from oxidative stressinduced damage. Appropriate increase of the activity of SIRT1 may be a therapeutic strategy to delay AMD.

Key words: H2O2, Cell growth, Silent information regulator of transcription 1, Retinal pigmented epithelium cells, Reactive oxygen species, siRNA, Oxidative stress

中图分类号: 

  • R774
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