Comparative Use of 2,7-dichlorofluorescin diacetate,dihydrorhodamine 123, and hydroethidine to study oxidative metabolism in phagocytic cells |
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| Authors: | A V Bizyukin L G Korkina B T Velichkovskii |
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| Institution: | (1) Institute of Pulmonology, Ministry of Health of Russian, Moscow;(2) Institute of Pediatric Hematology, Ministry of Health of Russian, Moscow;(3) Russian State Medical University, Moscow;(4) Institute of Pulmonology, 32/6 11-ya Parkovaya ulitsa, 105077 Moscow, Russia |
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| Abstract: | Tests carried out with model systems to assess the sensitivity of 2,7-dichlorofluorescin diacetate, dihydrorhodamine 123,
and hydroethidine to the reactive oxygen species generated in these systems showed that all three fluorochromes are sensitive
to hydroxyl and superoxide radicals and to hypochlorite ions and less sensitive to hydrogen peroxide. They do not react with
singlet oxygen. Hydroethidine reacts with superoxide radicals in the 1∶1 molar ratio. The relative proportions of reactive
oxygen species generated extra- and intracellularly were estimated for phorbol myristate acetate-stimulated peritoneal and
alveolar rat macrophages and for polymorphonuclear leukocytes from the blood of healthy donors. In activated leukocytes of
different types, intracellular generation accounted for approximately 20% of the total reactive oxygen species produced. Intracellular
production of these species was found to show considerable cyanide sensitivity, which indicates that cyanide-resistant NADPH
oxidase located in the plasma membrane is not implicated in the generation of reactive oxygen species.
Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 119, N
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4, pp. 361–365, April, 1995 |
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| Keywords: | 2 7-dichlorofluorescin diacetate dihydrorhodamine 123 hydroethidine oxygen radicals |
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