Detection of low levels of nitric oxide using an electrochemical sensor.
Detection of low levels of nitric oxide using an electrochemical sensor.
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DOI：
10.1007/978-1-61737-964-2_7
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Jo H
中科院分区：
文献类型：
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作者：
Boo YC;Mun GI;Tressel SL;Jo H
由一氧化氮合酶产生的一氧化氮介导了生物系统中的各种生理和病理事件。然而，对生物来源的一氧化氮进行定量评估仍然是一项艰巨的任务。在这里，我们描述了一个程序，用于定量低水平的一氧化氮使用一氧化氮选择性电化学传感器。一氧化氮被氧化为亚硝酸盐和/或硝酸盐，并在水介质中积累。首先，通过酶的方法将生物液体或培养基中的硝酸盐转化为亚硝酸盐。然后，亚硝酸盐在酸性碘化液中化学转化为等摩尔一氧化氮，其中一氧化氮由传感器检测。利用这种方法，本研究证明sirna介导的一氧化氮合酶3的抑制导致人脐静脉内皮细胞基础一氧化氮产量显著下降。ng -硝基- l -精氨酸甲酯也能抑制HUVECs的基础一氧化氮生成，而ng -硝基- d -精氨酸甲酯（D-NAME）则不能。本文提出的分析方法为测量生物源的基础和受刺激的一氧化氮生成提供了一种灵敏和方便的工具。
Nitric oxide produced from nitric oxide synthases mediates various physiological and pathological events in biological systems. However, quantitative assessment of nitric oxide from biological sources remains a difficult task. Here we describe a procedure for the quantification of low levels of nitric oxide using a nitric oxide – selective electrochemical sensor. Nitric oxide is oxidized to nitrite and/or nitrate and accumulated in the aqueous media. First, nitrate in biological fluids or culture media is converted to nitrite by an enzymatic method. Nitrite is then chemically converted to equimolar NO in an acidic iodide bath, where nitric oxide is detected by the sensor. Using this method, the present study demonstrates siRNA-mediated suppression of nitric oxide synthase 3 leading to a significant decline of basal nitric oxide production in human umbilical vein endothelial cells. Basal nitric oxide production from HUVECs is also shown to be inhibited by NG-nitro-L-arginine methyl ester but not by NG-nitro-D-arginine methyl ester (D-NAME). The analytical method presented here provides a sensitive and convenient tool for measuring basal and stimulated nitric oxide production from biological sources.