Assessment of oxidative DNA damage and repair at single cellular level via real-time monitoring of 8-OHdG biomarker
Assessment of oxidative DNA damage and repair at single cellular level via real-time monitoring of 8-OHdG biomarker
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DOI：
10.1016/j.bios.2010.08.029
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发表时间：
2010-12-15
影响因子：
12.6
通讯作者：
Li, Chen-Zhong
中科院分区：
文献类型：
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作者：
Prabhulkar, Shradha;Li, Chen-Zhong
8-羟基脱氧鸟苷(8-OHdG)是氧化应激最重要和最有文献记载的生物标志物，参与多种疾病的发生。8-OHdG水平与氧化DNA损伤有关，这是已知的各种与年龄相关的慢性病的根本原因。我们研究的目的是开发一种能够在单个细胞表面实时测量8-OHdG的检测策略。采用活性碳纤维微电极作为传感平台。用微电极检测尼古丁对单个肺上皮细胞释放8-羟色胺的影响。为了评价尼古丁在烟草遗传毒性中的直接作用，我们研究了尼古丁浓度和暴露时间等参数对8-OHdG分泌的影响。2-8 mM尼古丁溶液诱导单细胞DNA损伤呈剂量依赖性，这是通过电流测量分泌的8-OHdG生物标志物观察到的。4 mM尼古丁溶液对单个细胞的实时8-OHdG测定显示，110分钟后8-OHdG的分泌停止。我们已经成功地概述了一种在单细胞表面检测8-OHdG的方法。类似的方案可用于评估其他疾病模型中的氧化DNA损伤和修复机制。(C)2010爱思唯尔B.V.保留所有权利。
8-Hydroxydeoxyguanosine (8-OHdG) is the most important and best-documented biomarker of oxidative stress, which is involved in the instigation of various diseases. 8-OHdG levels correlate to oxidative DNA damage which is known to be the root cause of a variety of age-related chronic diseases. The purpose of our research was to develop a detection strategy capable of measuring 8-OHdG in real-time at the surface of a single cell. Activated carbon fiber microelectrodes were used as the sensing platform. The microelectrodes were used to measure 8-OHdG release from single lung epithelial cells under the influence of nicotine. In order to evaluate the direct role of nicotine in tobacco induced genotoxicity, we studied the influence of parameters such as nicotine concentration and exposure times on 8-OHdG secretion. 2-8 mM nicotine solutions induced dose-dependent DNA damage in single cells, which was observed via amperometric measurements of secreted 8-OHdG biomarker. Real-time 8-OHdG measurements from single cells exposed to 4 mM nicotine solution revealed cessation of 8-OHdG secretion after 110 min. We have successfully outlined a methodology to detect 8-OHdG at the surface of single cells. A similar protocol can be used to evaluate oxidative DNA damage and repair mechanisms in other disease models. (C) 2010 Elsevier B.V. All rights reserved.