Ashkenazi A, Abu-Rabeah K, Marks R S
Department of Biotechnology Engineering, Ben-Gurion University of the Negev, Beer-Sheva, Israel.
Talanta. 2009 Feb 15;77(4):1460-5. doi: 10.1016/j.talanta.2008.09.035. Epub 2008 Sep 30.
Several methodologies have been used in clinical chemistry for real-time assessment of NADPH oxidase primary product superoxide anion which dismutases to hydrogen peroxide. Among these methodologies, isoluminol chemiluminescence (CL) is considered to be one of the more sensitive and reliable techniques for the assessment of NADPH oxidase activity in neutrophils. The electrochemical technique was recently designed and also applied for real-time detection of NADPH oxidase activity in neutrophils but its reliability and sensitivity has not been investigated so far. In this study, isoluminol CL and electrochemical techniques were investigated and compared by monitoring the generation of superoxide and hydrogen peroxide in both PLB 985 cell line differentiated into neutrophil-like cells and human neutrophils. The electrochemical technique was shown to be as sensitive as that of CL and able to detect the reactive oxygen species (ROS) release of as low as 500 cells. Thus, the electrochemical technique could be used as an alternative to optical techniques for the evaluation of extracellular ROS in phagocyte cells.
临床化学中已采用多种方法来实时评估烟酰胺腺嘌呤二核苷酸磷酸（NADPH）氧化酶的主要产物超氧阴离子，该超氧阴离子会歧化为过氧化氢。在这些方法中，异鲁米诺化学发光（CL）被认为是评估中性粒细胞中NADPH氧化酶活性较为灵敏且可靠的技术之一。电化学技术是最近设计出来的，也用于实时检测中性粒细胞中的NADPH氧化酶活性，但到目前为止其可靠性和灵敏度尚未得到研究。在本研究中，通过监测分化为中性粒细胞样细胞的PLB 985细胞系和人中性粒细胞中超氧阴离子和过氧化氢的生成，对异鲁米诺CL和电化学技术进行了研究和比较。结果表明，电化学技术与CL技术一样灵敏，能够检测低至500个细胞释放的活性氧（ROS）。因此，电化学技术可作为一种替代光学技术的方法，用于评估吞噬细胞中的细胞外ROS。
