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J4 ›› 2013, Vol. 48 ›› Issue (7): 14-19.

• 前沿进展 • 上一篇    下一篇

贝特舒对人角膜上皮细胞凋亡诱导作用的实验研究

孙倩,樊廷俊*,邱月,葛源,于苗苗   

  1. 中国海洋大学 海洋生命学院角膜组织工程实验室, 山东 青岛 266003
  • 收稿日期:2013-03-13 发布日期:2013-12-03
  • 通讯作者: 樊廷俊(1964- ), 男, 博士,教授/博士生导师,研究方向为细胞工程与角膜组织工程. Email:tjfan@ouc.edu.cn
  • 作者简介:孙倩(1989- ), 女, 硕士研究生, 研究方向为细胞工程与角膜病. Email:sundarui0324@163.com
  • 基金资助:

    国家高技术研究发展计划(863计划)资助项目(2006AA02A132)

Experimental studies of the apoptosis-inducing effect of betaxolol on human corneal epithelial cells

SUN Qian, FAN Ting-jun*, QIU Yue, GE Yuan, YU Miao-miao   

  1. Laboratory for Corneal Tissue Engineering, College of Marine Life Sciences, Ocean University of China,
    Qingdao 266003, Shandong, China
  • Received:2013-03-13 Published:2013-12-03

摘要:

为了揭示常用青光眼药物贝特舒(betaxolol)对人角膜上皮(HCEP)细胞的毒性及其作用机理,使用不同质量浓度贝特舒对体外培养HCEP细胞进行了处理,并利用倒置显微镜跟踪观察了细胞的生长和形态,用吖啶橙/溴化乙锭(AO/EB)荧光双染色法检测了质膜的通透性,用琼脂糖凝胶电泳法检测DNA的断片化,用透射电镜检测细胞的超微结构。发现在0.17500~2.80000g/L的质量浓度范围内,贝特舒对HCEP细胞具有显著的毒性,并随着质量浓度的提高和处理时间的延长而逐渐增大,处理24h即可使大部分细胞皱缩死亡;进一步的研究结果发现,质量浓度为0.17500~2.80000g/L的贝特舒能显著提高HCEP细胞的质膜通透性,并使其出现DNA断片化、染色质凝缩和形成凋亡小体等凋亡细胞的结构变化。可见,在贝特舒0.17500~2.80000g/L的质量浓度范围内对HCEP细胞具有显著的毒性,并具有质量浓度和时间依赖性,其毒性作用的发挥是通过诱导细胞凋亡实现的,在眼科临床应用中毒副作用极大。

关键词: 人角膜上皮细胞;贝特舒;细胞凋亡;DNA断片化;凋亡小体

Abstract:

To examine the cytotoxic effect of betaxolol, a widely used anti-glaucoma drug, on human corneal epithelial (HCEP) cells and its underlying mechanisms, in vitro cultured HCEP cells were treated with betaxolol at different doses. And the growth situation and morphology of the cells were monitored with an inverted light microscope, plasma membrane permeability of them was detected by acridine orange/ethidium bromide (AO/EB) double-fluorescent staining, DNA fragmentation and ultrastructure of them were investigated by DNA agarose gel electrophoresis and transmission electron microscopy (TEM), respectively. It was found that betaxolol at doses of 0.17500-2.80000g/L showed significant cytotoxicity to HCEP cells. The cytotoxicity increased with dose and time, and most of the cells shrunk and died 24h post treatment. Furthermore, 0.17500-2.80000g/L betaxolol elevated the plasma membrane permeability of HCEP cells, induced DNA fragmentation, chromatin condensation, and apoptotic body formation as well. In conclusion, betaxolol at doses ranging from 0.17500 to 2.80000g/L has significant cytotoxicity to HCEP cells in dose-and time-dependent manners, which is realized by inducing apoptosis in these cells, suggesting the inescapable sever cytotoxic side effect of betaxolol in eye clinic.

Key words: human corneal epithelial cells; betaxolol; apoptosis; DNA fragmentation; apoptotic body

中图分类号: 

  • R772.2
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