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The use of methyphene and Vitamix Se in bulls of the E3 group helped reduce lipid peroxidation's final product. When comparing experimental groups with the control, it was found that on the fifth day of the experiment, the MD level decreased by 6, 6.5, and 7.6%, respectively. On the tenth day in the blood of the Ei group, the level was 0.263±0.0ii ^mol/l, in the E2 group, the level was 0.261±0.010 ^mol/l, in the E3 group, the level of MD was 0.251±0.011 pmol/l. On the fifteenth day of the experiment, the level of malonic di-aldehyde continued to decrease relative to the values of the C group. This figure felt in animals treated with methyphene - by i2%, in animals treated with Vitamix Se - by 13%, and in bulls treated with methyphene. Together with Vitamix Se reduced by i5%. On the twentieth and thirtieth days in the blood of bulls of all experimental groups again noted a decrease in the level of MD and reached the physiological values.
It should be noted that the combined use of methyphene and Vitamix Se in animals with chronic nitrate-nitrite toxicosis with cadmium loading contributed to a better reduction of the final products of lipid peroxidation.
Summing up the results of the research, we came to the following conclusions:
- the use of methyphene and Vitamix Se in the development of chronic nitrate-nitrite toxicosis of bulls with cadmium load contributed to the reduction of intermediate and final products of lipid peroxidation, namely diene conjugates and malonic dialdehyde;
- at nitrate-cadmium loading of bulls, the best effect on inhibition of processes of peroxidation of lipids of an organism of animals was shown by the combined use of metifen and Vitaminx Se.
References
1. Gildenskiold, R.S., Novikov, Yu.V., & Kha-miduli, R.S. (1992). Heavy metals in the environment and their effect on the body (review). Hygiene and sanitation, 5-6, 6-9.
2. Gonsky, Y.I., Yastremskaya, S.O., Boychuk, B.R. (2001). Age features of violation of lipid peroxidation and activity of energy-supplying enzymes in cadmium intoxication. Medical Chemistry, 3(1), 16-19.
3. Gutiy, B.V. (2005). The effect of sodium nitrate in a toxic dose on lipid peroxidation. Nauk. Visn. Lviv. nat. acad. vet. med. S.Z. Gzhytskoho, 7(2), 16-19.
4. Kmet, T.I., Vlasik, L.I. (2004). Features of the state of peroxide oxidation of proteins in juvenile animals with different types of metabolism under conditions of nitrate-cadmium intoxication. Modern problems of toxicology, 2, 36-38.
5. Slobodian, S. O., Gutyj, B. V., & Leskiv, K. Y.
(2019). The level of lipid peroxidation products in the rats blood under prolonged cadmium and lead loading. Ukrainian Journal of Veterinary and Agricultural Sciences, 2(3), 15-18. doi: 10.32718/ujvas2-3.04
6. Lavryshyn Yu.Yu., Gutyj B.V., Paladiychuk O.R. Influence of metisevit and lipointersil on morphological indices of bull blood under cadmium loading. Colloquium-journal, 2020, №18 (70), 10-14.
7. Lavryshyn Y., Gutyj B., Leskiv Ch., Kalyn B., Holub O., Romanovych M. The effect of Cadmium on the level of vitamins A and E in the blood of young cattle. Colloquium-journal, 2020, №32 (84), 11-15
8. Lesyk, Y., Ivanytska, A., Kovalchuk, I., Monas-tyrska, S., Hoivanovych, N., Gutyj, B., Zhelavskyi, M., Hulai, O., Midyk, S., Yakubchak, O., Poltavchenko,T.
(2020). Hematological parameters and content of lipids in tissues of the organism of rabbits according to the silicon connection. Ukrainian Journal of Ecology, 10(1), 30-36. doi: 10.15421/2020_5
9. Varkholiak I. S., Gutyj B. V., Kushnir V. I., Nazaruk N. V., Lisnyak O. I., & Yurynets T. V. (2020). The effect of bendamine on the intensity of lipid perox-idation and the activity of the antioxidant defense system of blood in rats in experimental doxorubicin-in-duced cardiomyopathy. Scientific Messenger of Lviv National University of Veterinary Medicine and Biotechnologies. Series: Veterinary sciences, 22(100), 3640. doi: 10.32718/nvlvet10007
Varkholiak I.S., Gutyj B. V., Leskiv Kh. Ya.,
Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies,
Lviv, Ukraine Kushnir V.I.,
State Scientific-Research Control Institute of Veterinary Medicinal Products and Feed Additives,
Lviv, Ukraine Hariv I.I., Martyshuk T. V., Guta Z.A.
Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies,
Lviv, Ukraine
DOI: 10.24412/2520-6990-2021-794-18-21 THE EFFECT OF BENDAMINE ON ANTIOXIDANT PROTECTION OF RATS' MYOCARDIUM IN
DOXORUBICIN INTOXICATION
Abstract.
The study aimed to investigate the effect of the remedy "Bendamine" on rats' antioxidant status in experimental doxorubicin-induced cardiomyopathy. The research was performed on white sexually mature young male Wistar rats weighing 180-200 g. Animals were divided into three groups of six rats each: control group - intact; experimental group E1 - doxorubicin was applicated intraperitoneally at a dose of 2.5 mg/kg 3 times a week for two weeks; experimental group E2 - in case of doxorubicin intoxication, the drug "Bendamine" was administered intragastrically at a dose of 20 mg/kg. The action of doxorubicin and the development of the hypoxic state in rats are accompanied by activation of oxidative stress and enhancement offree radical processes. It was indicated by
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increased levels of intermediate and end products of lipid peroxidation and inhibition of antioxidant defense. There was a 32.7% increase in diene conjugates and a 37.6% increase in TBA-active product in rats' control groups. After application to rats, it was also found that doxorubicin causes inhibition of the glutathione system of antiox-idant protection of animals. The use of the remedy "Bendamine" in the E2 group contributed to the strengthening of the enzymatic and non-enzymatic parts of the antioxidant system, protecting the structural and functional integrity of cell biomembranes. The medicine "Bendamine" inhibits the excessive formation of LPO products in pathologically altered tissues of rats' hearts. In the E2 group, the level of intermediate and final products is probably reduced. In the E2 group, the myocardial homogenate, the level of diene conjugates decreased by 16.8% also the level of TBA-active products - by 20.8% compared to the Ei group-had characteristic clinical signs of cardiomyopathy caused by doxorubicin.
Keywords: pharmacology, drug "Bendamine," doxorubicin, rats, cardiovascular insufficiency.
Introduction
Cardiovascular pathologies in dogs and cats are incredibly diverse and common in daily practice in Ukraine and abroad. According to the classification of Professor G.V. Domracheva, in domestic animals, cardiovascular pathology is divided into myocardial, pericardial, endocardial, and vascular diseases. There is a close relationship between the above pathologies, age, and animal breeds [4, 6, 10, 11].
Heart failure in dogs can be both a congenital disability and acquired as a result of infection. This pathology occurs mainly in elderly dogs. With heart failure in dogs, the heart cannot fully perform its physiological functions and provide the body with normal blood circulation. Accordingly, it leads to stagnation and deterioration of blood supply to organs, leading to pathological myocardium changes. [2, 3]. Analyzing domestic and foreign researchers' data, the development of the easy-to-use and safe complex cardiac drug, the use of which will increase the effectiveness of treatment of animals and a wide range of cardiovascular pathologies, is timely and relevant to research for veterinary medicine [5, 7, 8].
The study aimed to investigate the effect of the drug "Bendamine" on the antioxidant protection of rats' myocardium in experimental modeling of heart failure.
Material and methods of research
The study was performed on white sexually mature young male Wistar rats weighing 180-200 g, which were kept on the standard diet of the institute vivarium of the State Scientific-Research Control Institute of Veterinary Medicinal Products and Feed Additives. Throughout the research, the rats were kept on a balanced diet containing all the necessary components; the animal's drinking water was obtained without restrictions from glass drinking bowls with a volume of 0.2 liters.
Experimental studies were conducted by the drug-biological experiment requirements to select analogs, control, compliance with the same conditions of feeding and maintenance during the research, and accounting for the results.
Twenty-four male rats were selected to create a model of doxorubicin-induced cardiomyopathy. Animals were divided into three groups of 6 rats each: control group - intact; experimental group E1, in which rats
were simulated doxorubicin-induced cardiomyopathy by intraperitoneal administration of doxorubicin at a dose of 2.5 mg/kg 3 times a week for two weeks; experimental group E2, in which animals after injection of doxorubicin were intragastrically applicated the drug "Bendamine" at a dose of 20 mg/kg.
The content of LPO products - diene conjugates and TBA-active products, the activity of antioxidant enzymes - glutathione peroxidase and glutathione reductase, and the level of reduced glutathione, were determined by methods [9].
All animal manipulations were performed under the European Convention for the Protection of Vertebrate Animals Used for Experimental and Scientific Purposes (Strasbourg, 1986).
The analysis of research results was carried out using the software package Statistica 6.0. The student's t-test assessed the probability of differences. The results were considered plausible at P < 0.05.
Results and discussion
The obtained data indicate that doxorubicin's administration to the experimental groups is accompanied by an intensification of free radical oxidation processes in the myocardium. Thus, based on the conducted researches, the growth of intermediate and final products of LPO was established, namely the increase of diene conjugates by 32.7% and TBA-active products - by 37.6% compared with the control group of rats (Table 1). Oxidative damage to proteins leads to disruption of cardiomyocyte metabolism. Thus, under conditions of intensification of free radical oxidation, was suppress the antioxidant system's enzymatic link, thus intensifying oxidative stress in the myocardium [1]. The negative effect of oxidative-modified proteins in the cell is apparently because oxidized proteins act as a source of free radicals that deplete cellular antioxidants' supply.
An increase in the amount of TBA-active products in the myocardium of rats under the influence of doxo-rubicin indicates myocardial depletion of polyunsatu-rated fatty acids, which are the main substrate for lipid peroxidation. The myocardium loses the primary source of prostaglandin synthesis necessary for its normal functioning and plays an essential role in myocardial adaptation processes to stress.
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Table 1
Indicators of lipid peroxidation intensity in rat myocardial homogenate in experimental modeling of heart failure and action of the drug "Bendamine" (M ± m, n = 6)
T .. ^ Groups of animals Indicators ---
Control_Experimental 1_Experimental 2
Diene conjugates, ^mol/g 6.45 ± 0.13 8.56 ± 0.39* 7.12 ± 0.22* TBA-active products, mmol/g_30.79 ± 4.50_42.36 ± 2.11*_33.57 ± 2.47
After applying the drug "Bendamine" to rats of the E2 group, inhibition of lipid peroxidation processes under doxorubicin intoxication conditions was found. In the E2 group, the level of intermediate and final products is probably reduced. In the myocardial homoge-nate, the diene conjugates' level decreased by 16.8%. The level of TBA-active products - by 20.8% compared to the E1 group-had characteristic clinical signs of car-diomyopathy caused by doxorubicin.
Thus, the remedy "Bendamine" inhibits the excessive formation of LPO products in pathologically altered tissues of the heart of rats, has an inducing effect
on the antioxidant defense system, thus protecting the structural and functional integrity of cell biomembranes.
Studies have shown that doxorubicin causes inhibition of the glutathione system of rats' antioxidant defense in the Ei group. Thus, the level of reduced glutathione in the animals' myocardial homogenate of the Ei group probably decreased by 45.3%. In comparison, glutathione reoxidase and glutathione reductase activity decreased by 26.1 and 33.3%, respectively, compared with the rats' control group (Table 2).
Table 2
Indicators of the antioxidant defense system in rats' myocardial homogenate in experimental modeling of heart failure and the action of the drug "Bendamine" (M ± m, n = 6)
Groups of animals
Indicators Control Experimental 1 Experimental 2
Reduced glutathione, ^mol/g 0.53 ± 0.08 0.29 ± 0.05* 0.39 ± 0.05*
Glutathione peroxidase, nmol/min x mg protein 6.81 ± 0.79 5.03 ± 0.54* 6.37 ± 0.37
Glutathione reductase, ^mol/min x mg protein 0.57 ± 0.11 0.38 ± 0.12* 0.51 ± 0.10
The use of the remedy "Bendamine" in rats of the E2 group under doxorubicin cardiomyopathy conditions contributed to the activation of the glutathione link of the myocardium antioxidant system. The level of reduced glutathione in the myocardial homogenate of the E2 group ranged from 0.39 ± 0.05 ^mol/g.
After studying glutathione peroxidase activity in the experimental groups in the myocardium, it was highest in the E2 group. This enzyme activity was correspondingly higher by 26.6% relative to intoxicated rats not treated by the remedy "Bendamine".
Glutathione reductase activity was also higher in the E2 group, where it was 0.51 ± 0.10 ^mol/min x mg of protein. In contrast, this figure was 0.38 ± 0.12 ^mol/min x mg of protein in the E1 group.
Thus, based on studies of the effect of the drug "Bendamine" on the antioxidant system of the myocardium of experimental rats under doxorubicin intoxication, it was found that the newly developed remedy has a corrective effect under oxidative stress inherent in doxorubicin-induced cardiomyopathy. The studies' results enrich the pharmacological characteristics of the drug "Bendamine," indicate its rather pronounced protective effect on the myocardium in experimental dox-orubicin cardiomyopathy, and is a convincing proof of the feasibility of the drug in veterinary practice.
Conclusions
The action of doxorubicin and the development of the hypoxic state in rats are accompanied by activation of oxidative stress and enhancement of free radical processes, as indicated by increased levels of intermediate
and end products of lipid peroxidation, as well as inhibition of antioxidant defense.
The use of the drug "Bendamine" in rats of the E2 group contributed to the strengthening of the enzymatic and non-enzymatic parts of the antioxidant system, protecting the structural and functional integrity of cell biomembranes.
The remedy "Bendamine" inhibits the excessive formation of LPO products in pathologically altered tissues of rats' hearts.
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