Comparison of three analytical methods for superoxide produced by activated immune cells

被引:4
作者
Zhu, Zhixiang [1 ]
Guo, Ran [1 ]
Li, Yunqian [1 ]
Li, Shanshan [1 ]
Tu, Pengfei [1 ]
机构
[1] Beijing Univ Chinese Med, Sch Chinese Mat Med, Modern Res Ctr Tradit Chinese Med, 11 Beisanhuan East Rd, Beijing 100029, Peoples R China
基金
中国国家自然科学基金;
关键词
Superoxide; Immune cells; WST-8; Dihydroethidium; Lucigenin; RESPIRATORY BURST; OXIDATIVE STRESS; LUCIGENIN; CHEMILUMINESCENCE; HYDROETHIDINE; FLUORESCENCE; LUMINOL; PROBES; ANION;
D O I
10.1016/j.vascn.2019.106637
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
Superoxide plays a key role in normal immune function and inflammatory diseases. In order to evaluate normal immune function or screen inhibitors of superoxide production for treating inflammatory diseases, it is very important to detect superoxide with good accuracy, sensitivity, and flexibility. In present study, we investigated three analysis methods of superoxide, colorimetric assay by WST-8, fluorescence assay by dihydroethidium and chemiluminescence assay by lucigenin, compared their precisions, specificities, sensitivities and time curve characteristics in superoxide analysis, and then validate their values in the screening of anti-inflammatory compounds. The results reveal that three analysis methods of superoxide all have good precisions and high specificities but have different sensitivities and time curve characteristics, which suggest their different applications. In addition, they can all be used in the screening of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitors and anti-inflammatory compounds.
引用
收藏
页数:7
相关论文
共 23 条
  • [1] BIOLOGICAL DEFENSE MECHANISMS - PRODUCTION BY LEUKOCYTES OF SUPEROXIDE A POTENTIAL BACTERICIDAL AGENT
    BABIOR, BM
    KIPNES, RS
    CURNUTTE, JT
    [J]. JOURNAL OF CLINICAL INVESTIGATION, 1973, 52 (03) : 741 - 744
  • [2] A simplified hydroethidine method for fast and accurate detection of superoxide production in isolated mitochondria
    Back, Patricia
    Matthijssens, Filip
    Vanfleteren, Jacques R.
    Braeckman, Bart P.
    [J]. ANALYTICAL BIOCHEMISTRY, 2012, 423 (01) : 147 - 151
  • [3] Activation of neutrophil respiratory burst by fungal particles requires phosphatidylinositol 3-phosphate binding to p40phox in humans but not in mice
    Bagaitkar, Juhi
    Matute, Juan D.
    Austin, Anthony
    Arias, Andres A.
    Dinauer, Mary C.
    [J]. BLOOD, 2012, 120 (16) : 3385 - 3387
  • [4] Critical evaluation of the use of hydroethidine as a measure of superoxide anion radical
    Benov, L
    Sztejnberg, L
    Fridovich, I
    [J]. FREE RADICAL BIOLOGY AND MEDICINE, 1998, 25 (07) : 826 - 831
  • [5] Bindokas VP, 1996, J NEUROSCI, V16, P1324
  • [6] THE EFFECT OF THE NADPH OXIDASE INHIBITOR DIPHENYLENEIODONIUM ON AEROBIC AND ANAEROBIC MICROBICIDAL ACTIVITIES OF HUMAN-NEUTROPHILS
    ELLIS, JA
    MAYER, SJ
    JONES, OTG
    [J]. BIOCHEMICAL JOURNAL, 1988, 251 (03) : 887 - 891
  • [7] Guzik TJ, 2003, J PHYSIOL PHARMACOL, V54, P469
  • [8] LUCIGENIN CHEMILUMINESCENCE IN THE ASSESSMENT OF NEUTROPHIL SUPEROXIDE PRODUCTION
    GYLLENHAMMAR, H
    [J]. JOURNAL OF IMMUNOLOGICAL METHODS, 1987, 97 (02) : 209 - 213
  • [9] Lucigenin and coelenterazine as superoxide probes in mitochondrial and bacterial membranes
    Kervinen, M
    Pätsi, J
    Finel, M
    Hassinen, IE
    [J]. ANALYTICAL BIOCHEMISTRY, 2004, 324 (01) : 45 - 51
  • [10] Oxidative stress in angiogenesis and vascular disease
    Kim, Young-Woong
    Byzova, Tatiana V.
    [J]. BLOOD, 2014, 123 (05) : 625 - 631