Fluorescence and HPLC Detection of Hydroxyl Radical by a Rhodamine-Nitroxide Probe and its Application in Cell Imaging
Fluorescence and HPLC Detection of Hydroxyl Radical by a Rhodamine-Nitroxide Probe and its Application in Cell Imaging
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DOI：
10.1007/s10895-013-1329-0
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发表时间：
2014-03-01
影响因子：
2.7
通讯作者：
Guo, Yong
中科院分区：
文献类型：
--
作者：
Cao, Linying;Wu, Qingfeng;Guo, Yong
设计了罗丹明硝基氧探针来检测羟基自由基（中心点 OH），与其他活性氧（ROS）相比，该探针对中心点 OH 具有较高的选择性，并且对芬顿反应产生的中心点 OH 具有线性荧光响应。 HPLC-MS检测产物表明反应的主要产物为O-甲基羟胺，HPLC-UV/vis和HPLC-FLD测得的产物峰面积均随中心点OH浓度的增加而成比例增大。探索该探针在生物系统中的应用，追踪细胞在抑制线粒体呼吸链复合物I的鱼藤酮诱导的氧化应激条件下中心点·OH的产生，发现适量的鱼藤酮可以诱导正常人肝细胞（L02）和人肝癌细胞（HepG2）不同程度地产生中心点·OH。
A rhodamine nitroxide probe was designed to detect the hydroxyl radical (center dot OH), which presented high selectivity for center dot OH over other reactive oxygen species (ROS) and linear fluorescence response to center dot OH produced by Fenton reaction. The product was detected by HPLC-MS, indicating that the main product of the reaction was O-methylhydroxylamine and the product peak areas measured by HPLC-UV/vis and HPLC-FLD both enhanced proportionally with the increase of center dot OH concentration. The application of the probe in biological system was explored to trace the production of center dot OH in cells under oxidative stress condition induced by rotenone which can inhibit the mitochondria respiratory chain complex I and we found that appropriate rotenone may induce the normal human liver cells (L02) and human hepatoma cells (HepG2) to produce center dot OH at different degrees.