The role of macronutrients in the implementation of the corrective effect of low-mineralized water in experimental metabolic syndrome.

Background
Metabolic Syndrome (MS) is a widespread pathological condition, a group of interconnected metabolic disorders that contribute to the development of a number of severe noncommunicable diseases. Natural mineral waters of various compositions are widely used in the correction of MS. Their biological activity and specificity of action is associated with the presence of specific components (micronutrients, biologically active substances). At the same time, many researchers do not pay enough attention to the role of the biological activity of macro nutrients in low mineralized mineral waters.


Objective
To assess the corrective effect of macro-components of mineral waters based on a comparative analysis of changes in the structure of internal organs of white rats with the MS model, receiving mineral waters of the same balneological type, but with different quantitative composition of macro components.


Materials and methods
The material for histological and histochemical tests were the internal organs of male white rats with body weight 280-320 g. Animals were ranked into 4 groups: I - rats served as a control, ӀӀ - rats with a model of MS; ӀӀӀ and ӀV - rats received the mineral waters against the background of MS modeling. The mineral waters used in the work were characterized by an increased (in close amounts) content of organic substances (C org.), similar in chemical but different in quantitative macro-component composition. At the same time, mineral waters practically did not differ in terms of total mineralization. Modeling of MS was carried out for 60 days by replacing 10% fructose solution in drinking water bowls, introducing white bread crackers into the diet, and excluding green mass from the diet. In groups where animals were corrected for MS, mineral waters were administered with an intragastric tube with olive at a dose of 1% of body weight daily, starting from the 60th day of the experiment, for 12 days. Upon completion of the experiment on day 72, histological sections (which were stained with hematoxylin-eosin) were prepared from the extracted pieces of the heart, stomach, liver and kidneys tissues. The succinate-dehydrogenase (SDH) and lactate-dehydrogenase (LDH) activity were determined on the prepared cryostat sections. Using a light microscope, changes in the structure of the above organs and changes in the activity of oxidative-restorative enzymes were evaluated.


Results
The activity of oxidative-restorative enzymes was approaching normal; some manifestations of changes in water metabolism in the animal organism persisted. At the end of the use of both mineral waters, a significant decrease was found in the content of visceral fat around the intestines, renal capsule, under the pericardium, which correlated with a decrease in the animal's body weight and restoration of the fasting blood glucose level to the cereal level. The authors believe that the established features of the corrective effect of both mineral waters on changes in the structure of internal organs are explained not so much by the difference in the content and ratio of the bioactive component (C org.), but by differences in the content of macronutrients. In mineral waters obtained by rats of group III, the content of Cl- and SO4+2 ions is 2.30 and 3.67 times, and Ca2+ and Мg2+ ions are 2.75 and 4.57 more than in mineral waters, which received rats of group IV. However, the content of HCO3-, Na+, K+ is 1.14 and 1.30 times higher in the mineral waters obtained by rats of group IV.


Conclusions
The authors believe that the macronutrients involved in the implementation of MWs biological activity affect the metabolic background of the organism, which creates the bioactive element - C org. conditions for the performance of more intensive corrective action.

diet. In groups where animals were corrected for MS, mineral waters were injected with an intragastric tube with olive at a dose of 1% of body weight daily, starting from the 60th day of the experiment, for 12 days. Upon completion of the experiment on day 72, histological sections (which were stained with hematoxylineosin) were made from the extracted pieces of the heart, stomach, liver and kidneys. T he succinatedehydrogenase (SDH) and lactatedehydrogenase (LDH) activity were determined on the prepared cryostat sections.
Using a light microscope, changes in the structure of the above organs and changes in the activity of oxidative-restorative enzymes were evaluated. When modeling MS in rats, an increase in the amount of visceral fat and a significant increase in fasting glucose levels were noted. In the studied internal organs there were signs of dystrophic changes, high activity of oxidative-restorative enzymes, signs of impaired water metabolism. In animals of the ӀӀӀ group, the use of mineral water was accompanied by normalization of the myocardial structure. In other organs studied, there were preserved foci of residual manifestations of dystrophy against the background of a general restoration of the structure.
In animals of the ӀV group, upon completion of the mineral water intake, normalization of the structure of the stomach and myocardium and the preservation of diffuse manifestations of degeneration in the liver and kidneys were established. T he activity of oxidative-restorative enzymes was approaching normal; some manifestations of changes in water metabolism in the animal organism persisted. At the end of the use of both mineral waters, a significant decrease was found in the content of visceral fat around the intestines, renal capsule, under the pericardium, which correlated with a decrease in the animal's body weight and restoration of the fasting blood glucose level to the cereal level. T he authors believe that the established features of the corrective effect of both mineral waters on changes in the structure of internal organs are explained not so much by the difference in the content and ratio of the bioactive component (C org.), But by differences in the content of macronutrients.
In mineral waters obtained by rats of group III, the content of Cl-and SO4+2 ions is 2.30 and 3.67 times, and Ca2+ and Мg2+ ions are 2.75 and 4.57 more than in mineral waters, which received rats of group IV. However, the content of HCO3-, Na+, K+ is 1.14 and 1.30 times higher in the mineral waters obtained by rats of group IV. Conclusions. It can be assumed that the minerals involved in the implementation of the biological activity of mineral water, changing the metabolic background, on which the bioactive element of mineral water in the form of C org., Has a more intense corrective effect. Using a light microscope, changes in the structure of the above organs and changes in the activity of oxidative-restorative enzymes were evaluated. When modeling MS in rats, an increase in the amount of visceral fat and a significant increase in fasting glucose levels were noted. In the studied internal organs there were signs of dystrophic changes, high activity of oxidative-restorative enzymes, signs of impaired water metabolism. In animals of the ӀӀӀ group, the use of mineral water was accompanied by normalization of the myocardial structure. In other organs studied, there were preserved foci of residual manifestations of dystrophy against the background of a general restoration of the structure.
In animals of the ӀV group, upon completion of the mineral water intake, normalization of the structure of the stomach and myocardium and the preservation of diffuse In mineral waters obtained by rats of group III, the content of Cl-and SO4+2 ions is 2.30 Qeios, CC-BY 4.0 · Article, March 10, 2020 Qeios ID: OXTUY2 · https://doi.org/10.32388/OXTUY2 4/17 and 3.67 times, and Ca2+ and Мg2+ ions are 2.75 and 4.57 more than in mineral waters, which received rats of group IV. However, the content of HCO3-, Na+, K+ is 1.14 and 1.30 times higher in the mineral waters obtained by rats of group IV. Conclusions. It can be assumed that the minerals involved in the implementation of the biological activity of mineral water, changing the metabolic background, on which the bioactive element of mineral water in the form of C org., Has a more intense corrective effect.
Introduction. Metabolic syndrome (MS) -a common pathological condition, which is a group of interconnected metabolic disorders; high blood pressure, an increase in fasting levels of triglycerides and a decrease in fasting levels of high density lipoproteins; increased glucose obesity (excess primarily visceral fat), the development of insulin resistance [1,2,3,4]. T hese metabolic features contribute to the development of nonalcoholic fatty liver disease, type 2 diabetes mellitus, atherosclerosis, arterial hypertension, and cancer pathology [5,6]. According to the International Diabetes In the pathogenesis of MS, today, along with malnutrition, an essential role is given to the metabolism of minerals [8,9]. Moreover, as indicated in the available literature, mineral waters (mineral waters), due to the high bioavailability of the minerals contained in them, can effectively correct metabolic disturbances characteristic of MS [10,11,12]. Among the many ions contained in mineral waters, significant for the correction of MS, along with Са+2, Mg+2 consider ions НСО3-, Сl -, Nа+ [13,14,15]. It should be noted that in accessible publications the emphasis is on the influence of the above ions on metabolic parameters. At the same time, the state of metabolism is closely related to the state of the substrate of metabolic processes, that is, with the structural and functional characteristics of internal organs.
Since in the available literature we did not meet data on the effect of mineral waters of various compositions on the structural and functional state of internal organs during the development of MS, the goal of our work was formulated as follows.
Objective: on the basis of a comparative analysis of changes in the structure of internal organs of white rats with a model of the metabolic syndrome treated with mineral waters, to evaluate the corrective effect of macro-ions contained in these waters on these changes.
Materials and methods. T he material of this study was the data obtained during the study of white rats of males of the Vistar line of outbred breeding weight 240 -270 g.
Certified animals obtained from the kennel of the emergency "Biomodelservice" GI "Pharmacology and toxicologists of NAMS of Ukraine", Kyiv.
T he work with the animal was carried out in accordance with the requirements of the European Parliament Directive (2010) and the Order of the Ministry of Education and Science, Youth and Sports of Ukraine (2012).
In accordance with the objectives of the work, animals were ranked in 4 groups.
Ӏ group -15 intact rats, kept under vivarium conditions, but were not exposed to any effects. T he data obtained during their study served as a control.
ӀӀ group -10 rats in which metabolic syndrome was simulated for 60 days.
ӀӀӀ group -10 rats, which, against the background of MS modeling, received mineral water of sample No. 1.
ӀV group -10 rats, which against the background of the simulation of MS received mineral water of sample No. 2.
Modeling of the metabolic syndrome was carried out for 60 days by replacing drinking water with a 10% solution of fructose and introducing white bread crackers into the standard diet. In groups where animals were corrected for MS, mineral waters were injected with an intragastric tube with olive at a dose of 1% of body weight daily, starting from the 60th day of the experiment, for 12 days. Weighed animals weekly. At the end of the experiment on day 72, the animals were removed from the experiment by decapitation under ether anesthesia. At an autopsy, after a visual assessment of the state of internal organs, pieces of the heart, stomach, liver, and kidneys were taken in a volume of 1 cm3. A piece was divided into two parts. One part was fixed in 4% steam with formaldehyde for 48 hours, then it was passed through alcohols of increasing concentration and poured into celloidin according to the generally accepted method.
Sections 7-9 μm thick were made from the obtained blocks, which were stained with hematoxylin-eosin. T he resulting preparations were examined under a light microscope to determine structural changes in the internal organs. T he second part of the piece was frozen with dry carbon dioxide (t = -44 ° C). Cryostat sections 11 μm thick were made from the obtained blocks, on which the activity of succinatedehydrogenase (SDH) and lactatedehydrogenase (LDH) were determined from the Lloyd K. prescriptions; enzyme activity was evaluated by the semi-quantitative method, the data were given in arbitrary units of optical density (conventional units) [18,19]. T he concentration of glucose in the blood was determined on an empty stomach using the glucose oxidant method [19,20]. active components and compounds in both waters is also below the balneological norm [21]. In general, we can assume that in terms of their physicochemical composition and biological activity these waters are fairly close.
Results and its discussion. Before proceeding to the presentation of the research results, the group with MS. In rats of group III, it increased from (293.6 ± 1.13) g to (343.2 ± 1.02) g, and in animals of group ӀV, body weight increased from (299.8 ± 1.15) g to ( 349.5 ± 0.87) g.   Conclusions. Summarizing the results of the study, it can be argued that mineral waters containing organic substances as the main bioactive agent have a positive effect on the structural characteristics of the internal organs of rats with the MS model. T his positive effect was manifested in a decrease or absence of signs of dystrophic processes. It should be noted that there were differences in the prevalence of these positive changes.
In both cases, after the application of these mineral waters in the myocardium, no manifestations of dystrophy were detected. In the stomach wall, residual manifestations of dystrophy when applying mineral water No. 1 were noted in the glands of the surface layers of the gastric mucosa, and when using mineral water No. 1 they were not detected. In the liver parenchyma, residual manifestations of dystrophic processes were noted in the cases of using both mineral waters, however, in the case of using mineral water No. 1, they are determined in the parenchyma of part of the lobules, and in the case of using mineral water No. 2 in all segments.
Accordingly, in the kidneys in both cases, the use of mineral water in the capillaries of the glomeruli of the renal corpuscles retains residual manifestations of dystrophic changes, but in the tubules when using mineral water No. 2 they are more common. In our opinion, it is incorrect to associate the revealed differences with different amounts of the bioactive component, since according to the data in T able 2, the difference does not Since SO4+2 ions affect the metabolic processes in hepatocytes, it can be assumed that their higher content in mineral water No. 1 determines the features of structural changes in the liver; the higher Cl-content in the same mineral water obviously causes a difference in the structural structure of the renal tubules. Differences in the content of Ca2+ and Mg2+, H2S, and H2SiO3 ions -affecting the course of metabolic processes and hemodynamic characteristics, obviously also affect the revealed differences in the structural changes in the internal organs of experimental rats.
T hus, the results of the studies indicate that, when assessing the biological activity of mineral waters of low mineralization (up to 1 g / l), it is necessary to take into account not only the presence and quantity of a specific agent (microelements and biologically active substances), but also the presence and quantitative ratio of macroelements.