Open AccessCelestine blue B as a sensor for hypochlorous acid and HOCL-modified proteins registration
Author(s) -
Veronika E. Lutsenko,
D. V. Grigorieva,
И. В. Горудко,
С. Н. Черенкевич,
N. Gorbunov,
В. А. Костевич,
О. М. Панасенко,
Sokolov Av
Publication year - 2019
Publication title -
medical academic journal
Language(s) - English
Resource type - Journals
eISSN - 2687-1378
pISSN - 1608-4101
DOI - 10.17816/maj19263-71
Subject(s) - hypochlorous acid , chemistry , biochemistry
Objective — the study of hypochlorous acid (HOCl) and its derivatives production, which catalyzed by human neutrophil myeloperoxidase, using “turn-on” fluorescent sensor — celestine blue B.
Materials and methods. Neutrophils were isolated from the venous blood of healthy donors. Phorbol 12-myristate 13-acetate, N-formyl-methionyl-leucyl-phenylalanine, plant lectins, HOCl-modified proteins were used as agonists. N-acetylcysteine, 4-aminobenzoic acid hydrazide, isoniazid and ceruloplasmin were used as regulators of neutrophil myeloperoxidase activity and/or HOCl scavengers.
Results. Using a wide range of agonists and inhibitors, it has been shown that celestine blue B is oxidized in vitro by HOCl and its derivatives as a result of neutrophil myeloperoxidase activity. The oxidation of celestine blue B by HOCl-modified human serum albumin (HSA-Cl) and inhibition of this process by monoclonal antibody against HSA-Cl (IgM class) was also found.
Conclusion. Based on the developed method using celestine blue B, it is possible to conduct a sensitive analysis for the presence of HOCl-modified proteins (chloramines, etc.), to investigate the effect of various agonists and drugs on myeloperoxidase activity and exocytosis from the neutrophil granules.