Department of Bionanotechnology, Gachon University, Sujeong-Gu, Seongnam-Si, Gyeonggi-Do, Republic of Korea.
Anal Chem. 2013 May 21;85(10):4902-11. doi: 10.1021/ac303068t. Epub 2013 May 3.
This Article describes an unprecedented, simple, and real-time in vitro analytical tool to measure the luminous effect on the time responses function of retinal ganglion cells (RGC-5) by electric cell substrate impedance sensing (ECIS) system. The ECIS system was used for the continuous measurement of different color light-induced effects on the response of cells that exposed to protective drugs. The measurement suggests that the association of photo-oxidative stress was mediated by reactive oxygen species (ROS), which plays a critical role that leads to cell stress, damages, and retinopathy, resulting in eye degenerative diseases. Continuous light radiation caused time-dependent decline of RGC-5 response and resulted in photodamage within 10 h due to adenosine 5'-triphosphate depletion and increased ROS level, which is similar to in vivo photodamage. The ECIS results were correlated with standard cell viability assay. ECIS is very helpful to determine the protective effects of analyzed drugs such as β-carotene, quercetin, agmatine, and glutathione in RGC-5 cells, and the maximum drug activity of nontoxic safer drug concentrations was found to be 0.25, 0.25, 0.25, and 1.0 mM, respectively. All drugs show protection against light radiation toxicity in a dose-dependent manner; the most effective drug was found to be glutathione. The proposed system identifies the phototoxic effects in RGC-5 and provides high throughput drug screening for photo-oxidative stress during early stages of drug discovery. This study is convenient and potential enough for the direct measurements of the photoprotective effect in vitro and would be of broad interest in the field of therapeutics.
本文描述了一种前所未有的、简单的、实时的体外分析工具,通过电细胞基质阻抗传感(ECIS)系统测量视网膜神经节细胞(RGC-5)时间响应功能的发光效应。ECIS 系统用于连续测量不同颜色光对暴露于保护药物的细胞反应的影响。测量结果表明,光氧化应激的关联是由活性氧(ROS)介导的,ROS 在导致细胞应激、损伤和视网膜病变,从而导致眼部退行性疾病中起着关键作用。持续的光辐射会导致 RGC-5 反应随时间的依赖性下降,并在 10 小时内由于三磷酸腺苷耗竭和 ROS 水平升高而导致光损伤,这与体内光损伤相似。ECIS 结果与标准细胞活力测定相关。ECIS 非常有助于确定β-胡萝卜素、槲皮素、胍丁胺和谷胱甘肽等分析药物对 RGC-5 细胞的保护作用,并且发现非毒性更安全药物浓度的最大药物活性分别为 0.25、0.25、0.25 和 1.0 mM。所有药物均以剂量依赖性方式显示对光辐射毒性的保护作用;发现最有效的药物是谷胱甘肽。该系统识别 RGC-5 中的光毒性作用,并在药物发现的早期阶段提供针对光氧化应激的高通量药物筛选。该研究对于体外光保护效果的直接测量非常方便且具有潜力,并且在治疗领域具有广泛的兴趣。