Application of Celestine Blue B and Gallocyanine for Studying the Effect of Drugs on the Production of Reactive Oxygen a
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Journal of Applied Spectroscopy, Vol. 87, No. 4, September, 2020 (Russian Original Vol. 87, No. 4, July–August, 2020)
APPLICATION OF CELESTINE BLUE B AND GALLOCYANINE FOR STUDYING THE EFFECT OF DRUGS ON THE PRODUCTION OF REACTIVE OXYGEN AND HALOGEN SPECIES BY NEUTROPHILS V. E. Reut,a* D. V. Grigorieva,a I. V. Gorudko,a A. V. Sokolov,b,c,d and O. M. Panasenkod,e
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We investigated the effect of drugs (dapsone, paracetamol, and isoniazid) on the production of reactive oxygen and halogen species by neutrophils. The standard fluorescent method using scopoletin as well as recently developed fluorescent methods using oxazine dyes, celestine blue B, and gallocyanine were employed for this purpose. Celestine blue B reacts selectively with hypochlorous acid, while gallocyanine reacts mainly with the superoxide radical-anion, which reveals the regulatory effect of anti-inflammatory drugs on neutrophil NADPH-oxidase and myeloperoxidase responsible for the production of reactive oxygen and halogen species, respectively. These results indicate that gallocyanine and celestine blue B hold promise as chemosensors for studying the effect of drugs used in antiinflammatory therapy for the neutrophil respiratory burst. Keywords: paracetamol, dapsone, isoniazid, neutrophil activation, oxidative stress, reactive oxygen species, reactive halogen species. Introduction. Neutrophils (PMNs, polymorphonuclear leukocytes) are effector cells of inherent immunity and comprise up to 70% of the total amount of leukocytes circulating in the blood. PMNs serve as the first line of defense against the invasion of pathogens. These cells are rapidly activated upon contact with pathogen-associated molecular patterns such as lipopolysaccharides and formyl-containing bacterial polypeptides. The reaction of PMNs entails chemotaxis to the site of inflammation, phagocytosis, the formation of active species of oxygen (ROS) and halogens (RHS), degranulation, and the release of extracellular traps (NETosis) [1, 2]. The ROS are produced by an NADPH-oxidase complex generating the superoxide radical-anion ( i O −2 ), which is converted into hydrogen peroxide (H2O2) either spontaneously or with the participation of superoxidedismutase. The heme-containing enzyme of azurophilic granules of PMNs, namely, myeloperoxidase (MPO), catalyzes the oxidation of chloride by hydrogen peroxide to give hypochlorous acid (HOCl). The reaction of HOCl with biopolymers of bacteria and lower fungi leads to their annihilation, i.e., is a significant part of the antimicrobial activity of PMNs [3–5]. The ability to produce ROS, RH, and their derivatives as well as vasoactive lipid mediators permits PMNs to participate in the inflammatory reaction and modulate the immune response [4, 6, 7]. However, the powerful action of oxidizing agents on pathogens is not selective. Thus, the accumulation of ROS and RHS at a site of inflammation leads to the peroxide oxidation of lipids (damage to cell membranes and lipoproteins), oxidative damage to proteins, DNA mutations, an
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