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10«<g©yL©(MUM~JOy©MaL» #794)), 2©21 / AGRICULTURAL sciences
27
AGRICULTURAL SCIENCES
UDC 606:637.3:637.055
Kushnir 1.1., Tsisaryk O. Y.
Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies,
Lviv, Ukraine
DOI: 10.24412/2520-6990-2021-794-27-29 ANTIOXIDANT SYSTEM OF RATS BY THE INFLUENCE OF COMPOSITION OF BACTERIA ISOLATED FROM TRADITIONAL CARPATHIAN CHEESE BRYNDZA
Abstract.
The article presents data on the study of the effect of the composition of probiotic strains of L. lactis, Lb. plantarum and E. durans SB18 in a ratio 50:40:10 on some indicators of the antioxidant defense system of white rats. In particular, the use ofthe tested composition caused an inhibitory effect on the intensity of lipid peroxidation processes, which in turn led to decrease of TBA-active products and lipid hydroperoxides. It was found that on the 14th day of the experiment the content of TBA-active products and lipid hydroperoxides decreased by - 6.1 and 4.1% respectively and on the 21st day of the experiment - by 14.9 and 22.1% (p<0.01).
In determining the content of products of oxidative modification of proteins (OMP), which characterize the oxidative damage of various components of the cell, a slight decrease in the concentration of OMP430 and OMP370 was noted throughout the experiment. Under these conditions, was established the activation of metabolic processes, which indicated by an increase in superoxide dismutase activity (SOD) on 14 and 21 day of the experiment by 6.3 and9.6% respectively (p<0.05).
In addition, an increase of humoral defense factors of laboratory animal was found, in particular on the 21st day of the experiment the serum lysozyme activity increased by 9.1%, and the level of circulating immune complexes (CICs) decreased by 14%.
Keywords: antioxidant defense system, E. durans, L. lactis, Lb. plantarum, lipid peroxidation processes, TBA-active products
Introduction
Contravention of the balance between endogenous and pathogenic microflora causes functional disorders of the digestive system, which can be the reason of disorders of psycho-emotional state and an increased sensitivity to the stress [1]. Given this, dysbacteriosis and stress conditions mutually reinforce each other, which generally causes a violation of homeostasis of the macroorganism. In this case, in the body occurs excessive immunoreactivity, and in the tissues activation of lipid peroxidation and also when the functional interaction between the immune and hypothalamic-pituitary systems is disrupted [2, 3]. It is confirmed, that under stress conditions the processes of peroxide oxidation in the body intensify [4].
It was found, that one of the mechanisms of the action of probiotics is their restoration of pro- and antioxidant balance. The obtained results indicated on the effectiveness of combined probiotics in the prevention of stress-induced lesions [5-7]. The therapeutic effect of lactic acid bacteria is also provided with their rapid colonization of the gastric mucosa [8].
Considering the literature data about the antioxidant properties of probiotics [9], the studies were conducted to determine their effect on the lipoperoxidation processes, in particular were found decrease in the concentration of DC after 14 days of administration of combination of lactic acid bacteria, and also was observed decrease of TBA-active products and catalase activity in comparison to the control group of rats [5, 10].
For more detailed study of the effectiveness of new probiotic strains of microorganisms a topical issue
is research directed on studying the antioxidant protection of the macroorganism. So the aim of this work was to study the effect of the developed composition of lactic acid bacteria [11] which were isolated from traditional Carpathian cheese bryndza [12] on some indicators of the antioxidant protection system of white rats.
Materials and methods.
Determination of the influence of the studied composition of the consortium of lactic acid bacteria on the antioxidant defense system was performed on white rats, Wistar line, body weight 160-180 g. For this purpose, was formed control and experimental group of animals, with six animals in each group. Animals of the first (control) group were administered isotonic sodium chloride solution, animals of the second group - a composition of probiotic strains of L. lactis, Lb. plantarum and E. durans SB18 in a ratio of 50:40:10 at a dose of 1 cm3 (M07 CFU/cm3) per animal. The tested compositions were administered intragastrically by using a metal probe for 21 days. After that, under light ether anesthesia, the animals were decapitated and blood and tissues were taken for research. The level of intensity of oxidative destruction of proteins was evaluated by the reaction of the obtained carbonyl derivatives of the amino acid reaction with dinitrophenylhydrazine, as described in Levine et al. [13]. Lysozyme activity of blood serum was determined by nephelometric method [14]. Determination of the content of circulating immune complexes (CICs) in the serum was performed according to the method of Grinevych Yu. A. et al. [15]. The concentration of TBA-active products in blood plasma was determined by the method of Korobeiny-kov E. N. [16].
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AGRICULTURAL SCIENCES / «coyyomum-jmtmal» 2©2fl
Determination of superoxide dismutase activity was performed according to the method of Dubynyna E. E. et al. [17]. The content of lipid hydroperoxides in plasma blood was determined by the method described by Myronchyk V. V. [18].
Results and discussion.
The use of a composition of lactic acid bacteria L. lactis, Lb. plantarum and E. durans SB18 throughout the experiment caused an inhibitory effect on the inten-sivity of lipid peroxidation processes, which led to the decrease of the content of TBA-active products and lipid hydroperoxides (Table 1). In particular, on the 14th
An extremely important enzyme of antioxidant protection of the body is superoxide dismutase (SOD), which is the primary line of defense against oxidative damage and provides interruption of chains in oxygen-dependent of free-radical reactions [20].
In the determining the activity of SOD (Table 2) it was found out, that the use of the studied composition of lactic acid microorganisms for 14 days caused its increased by 6.3%, and on the 21st day of the experiment this figure was probably higher by 9.6% (p<0.05). Therefore, an increase of SOD activity may indicate on the activation of metabolic processes, in particular, an increased demand for O2, and an increased generation of active form of oxygen.
Note: ** - p<0.05
In determining the influence of the studied composition on humoral defense factors (Table 3), it was found, that on the 14th day of the experiment, the lyso-zyme activity of blood serum increased only by 0.8%. At the same time, there was a decrease the level of CICs by 4.9% compared to the values of the control group.
day of the administration of consortium of lactic acid bacteria the content of TBA-active products and lipid hydroperoxides decreased by 6.1 and 4.1%, respectively, and on the 21st day - by 14,9 and 22.1% (p<0.01 ). Because lipid peroxidation is a physiological process and represet the degree of influence of molecular oxygen on mitochondrial lipids under normal physiological conditions and is determined by the ability to regulate the structural and functional state of membranes [19], the detected changes may indicate on enhanced antiox-idant and detoxification properties of lactic acid bacteria.
The evaluation of oxidative damage of various components of the cell was carried out on the content of products of oxidative modification of proteins (OMP). It should be noted that OMP is one of the earliest and most reliable indicators of tissue damage, as OMP derivatives are stable [21, 22, 23]. That is why we studied the effect of the studied composition of lactic acid bacteria on OMP. As can be seen from the data in Table 2, long-term administration of a consortium of lactic acid bacteria caused a slight decrease in OMP430 and OMP370, which is likely to indicate an inhibitory effect of the test composition on the processes of OMP.
Table 2
Whereas, on the 21st day of the experiment, the Lyso-zyme activity an increased by 9.1%, and the level of the CICs decreased by 14%. The detected changes may indicated about an increase in humoral defense factors and normalization of homeostasis.
Table 3
Humoral defense factors in blood serum of rats
Parameters 14 day 21 day
Control Experiment Control Experiment
Lysozyme activity, % 35.7±2.33 36.0±1.15 28.7±0.88 31.3±1.45
CICs, mmol/L 41.0±1.0 39.0±3.46 45.0±3.51 38.7±1.86
Table 1
Indicators of lipids peroxidation in plasma blood of rats
Parameters 14 day 21 day
Control Experiment Control Experiment
TBA-active products, nmol/ml 4.459±0.29 4.184±0.32 3.966±0.40 3.374±0.09
Lipid hydroperoxides, OdE/ml 2.18±0.08 2.09±0.12 2.26±0.06 1.76±0.012**
Note: ** - p<0.01
SOD activity and content of products of oxidative modification of proteins
Parameters 14 day 21 day
Control Experiment Control Experiment
SOD (U/ml) 4.13±0.26 4.39±0.13 4.18±0.11 4.58±0.03*
OMP 430 (^M/ml) 3.68±0.13 3.36±0.10 3.57±0.11 3.51±0.09
OMP 370 (^M/ml) 0.78±0.021 0.75±0.029 0.79±0.025 0.75±0.017
«coyyomum-jmtmal» #794)), 2©21 / AGRICULTURAL sciences
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Conclusions.
1. The use of a consortium of lactic acid bacteria L. lactis, Lb.plantarum and E. durans SB18 in a ratio 50:40:10 caused decrease in the content of lipid peroxidation products, TBA-active products, the content of OMP products, which indicated to the an increase in antioxidant properties of the body.
2. Lactic acid bacteria of the formed composition had a positive effect on humoral defens factors of the organism of laboratory animals, in particular on the 21st day of its use lysozyme activity of blood serum increased by 9.1%.
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