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ECPB 2020, 89(1): 15–22
https://doi.org/10.25040/ecpb2020.01.015
Research articles

Characteristics of antioxidant and NO-synthase systems of blood lymphocytes under the action of decamethoxine

I.V. KOVALENKO, O.K. ONUFROVYCH, R.V. FAFULA, O.V. MELNYK, Z.D. VOROBETS
Abstract

Antiseptic drugs have mostly microbicidal effect on the skin surface, mucous membranes, surfaces of a wound and body cavities. Decamethoxine is a chemical compound [1, 10-decamethylene bis (N, N – dimethylmethoxycarbonylmethyl) ammonium dichloride] that has a wide range of antimicrobial actions on Gram-positive (staphylococcus, streptococcus, pneumococcus), Gram-negative (gonococcus, meningococcus) cocci, diphtheria corynebacteria, enterobacteria, pseudomonads, protozoa, dermatophytes, yeasts-like fungi of the genus Candida, chlamydia and viruses. Pathological processes that occur in the cells of the body when inflammatory processes and pathological conditions are caused primarily by the violation of regulatory systems, the activation of lipid peroxidation processes and the violation of antioxidant protective system. Since decamethoxine has hydrophilic and lipophilic properties, it can penetrate into cells, blood through surfaces of a wound, mucous membranes, skin, etc., and be transmitted by blood to different organs and tissues, thus causing a variety of biochemical effects, particularly regarding to the regulatory NO-synthase system of cells. In this respect the biological action of decamethoxine has not been investigated substantially. Since the intracellular lymphocyte metabolism is based on the physiologically- and biochemically- fixed ability of these cells to respond quickly to any changes in homeostasis in the body, they can be a convenient and adequate model for the study of pathophysiological and regulatory disorders in the organism. We have conducted a comparative study of the LPO system and glutathione in peripheral blood lymphocytes under the action of decamethoxine. A slight inhibition of LPO processes under the action of different concentrations of decamethoxine is shown which was evaluated by determining the concentration of malondialdehyde, being a secondary product of lipid peroxidation. Thus, the concentration of MDA in blood lymphocytes is (62,3±4,6) nmol/mg of protein in the control group, while under the influence of different concentrations of decamethoxine (10-5 – 10-2 M) this value decreases to (55,1±4,3) μmol/mg of protein (p>0,001). Significant changes in blood cells in the process of lipid peroxidation have been observed when they are not exposed to decamethoxine, however, there is a tendency to decrease. Simultaneously, with a slight decrease in the process LPO, the appropriate changes in the activity of glutathione system enzymes were revealed. Thus, it was shown that lymphocyte glutathione peroxidase activity is (154,2±13,4) nmol GSH/min∙mg of protein in the control group. Under the action of decamethoxine, this activity increases dose-dependently and reaches 181,2±13,3) nmol GSH/min∙mg of protein (p < 0,05). However, the concentration of reduced glutathione under the action of decamethoxine remained virtually unchanged and ranged from 17,2 to 18,4 nmol GSH/mg of protein. Glutathione S-transferase activity was (112,2±9,2) nmol GSH/min∙mg of protein in the control group. While being added to the incubation medium of decamethoxine at concentrations of 10-5−10-2M, this enzyme was activated dose-dependently, its activity increased to (156,6±12,4) nmol GSH/min∙mg of protein, that is in 1,4 times (p<0,05). The effect of decamethoxine on the activity of the arginase-NO-synthase system of blood lymphocytes was also demonstrated. Decamethoxine dose-dependent activated arginase activity up to (175,1±8,8) nmol urea/min∙mg of protein at 10-2 M concentration, that is 1,4 times relatively to the control group (p<0,05). It has been established that the cNOS activity of blood lymphocytes in the control group was 71,4±6,9 and the activity of the inducible isoform 1,58±0,18 nmol NADPH(H+)/min∙mg of protein. With the action of decamethoxine at a concentration of 10-5 M, the activity of cNOS decreased in 1,25 times (p<0,05) while the activity of iNOS increased in 10,8 times (p<0,001). Thus, it has been found that decamethoxine significantly influences the regulatory mechanisms of the cell, blood lymphocytes in particular. It activates the enzymes of the glutathione antioxidant system, stimulates the activity of arginase and inhibits the activity of the constitutive isoform of NO synthase with the simultaneous activation of the inducible isoform of NO synthase.

Recieved: 26.02.20

Keywords: decamethoxine, lymphocytes, glutathione, arginase, NO-synthase

Full text: PDF (Eng) 340K

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