Corresponding author: Andrii Demkovych ( demkovushae@tdmu.edu.ua ) Academic editor: Georgi Momekov
© 2021 Andrii Demkovych, Vitaliy Shcherba, Olha Yaremchuk, Halyna Stoikevych, Volodymyr Machogan, Vitalii Luchynskyi.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Demkovych A, Shcherba V, Yaremchuk O, Stoikevych H, Machogan V, Luchynskyi V (2021) Effects of flavonol quercetin on syndrome of endogenous intoxication in experimental periodontitis. Pharmacia 68(3): 627-632. https://doi.org/10.3897/pharmacia.68.e67341
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The article presents the results of study effect of quercetin on the indicators of endogenous intoxication syndrome under conditions of experimental periodontitis. One of the key links in the pathogenesis of any inflammatory disease, including maxillofacial area, is endogenous intoxication, an important pathophysiological mechanism of which is the activation of lipid peroxidation. The experimental animals were treated by intramuscular injections with water-soluble quercetin for 7 days. For further research the blood serum was selected in which was determined the content of middle molecular weight molecules and erythrocyte intoxication index. Excessive lipoperoxidation was accompanied by accumulation of peroxidation products and depletion of antioxidant reserves, which led to accumulation of toxic substances. That is why the study of the level of middle molecular weight molecules and erythrocyte intoxication index in simulated bacterial-immune periodontitis is a reliable criterion. Quercetin was able to reduce the expression of endogenous intoxication syndrome in experimental bacterial-immune periodontitis.
Periodontitis, endogenous intoxication, middle molecular weight molecules, erythrocyte intoxication index, lipid peroxidation, qercetine
The periodontal complex is the tissue that surrounds the tooth and ensures its stable position relative to the alveolar hole. It consists of gums, periodontal ligament, root cementum, bone tissue of alveolar processes. Periodontal disease affects either any of these components, or the entire periodontium. By nature, the process of change is dystrophic, tumor or inflammatory, and the latter type accounts for 90–93% of all diseases in this area. According to the WHO, 98% of the adult population of the planet has problems associated with periodontal tissue pathology. For many years there has been a tendency to earlier onset of this disease and its aggressive course (
Endogenous intoxication is the accumulation of endogenous toxic products in the body (
In this regard, the problem of timely diagnosis and monitoring of endogenous intoxication syndrome and correction of its main indicators in periodontology today is relevant and promising.
Objective. To study the indicators of endogenous intoxication syndrome and the effect of quercetin on them under the conditions of experimental periodontitis.
The experiment was conducted on 26 white outbred rats weighing 180–200 g, which were contained in the usual diet of vivarium. The research was performed according to the general rules and regulations of the European Convention for the Protection of vertebrate animals that use for experimental and other scientific purposes (Strasbourg 1986), the General ethical animal experimentation (Kyiv 2001). The animals were divided into three groups: 1st group – intact (10 rats); 2nd group – animals in which the model periodontitis on the 14th day (8 rats); 3rd group – animals in which the model periodontitis on the 14th day which was introduced flavonol quercetin (8 rats). The experimental bacterial-immune periodontitis in the experimental animals was caused by injection into the tissue of the periodontal complex of the microorganisms mixture diluted with egg`s albumin (
To determine the content of middle molecular weight molecules from the serum was isolated acid-soluble fraction, which was obtained by adding 1.8 ml of 10% solution of trichloroacetic acid to 0.2ml of serum. Subsequent centrifugation was performed at 3000 rpm for 30 minutes The isolated 0.5 ml fraction was diluted 1:10 with distilled water and the optical density was determined at 254 nm (chain amino acids) and 280 nm (aromatic amino acids) against distilled water on a spectrophotometer. The results were expressed in conventional units, numerically equal to the extinction (
The method of determining the erythrocyte intoxication index (EII) was based on information about erythrocytes as a universal adsorbent, which allows to estimate the level of EI by changing the sorption capacity of erythrocytes polar, almost impermeable to their methylene blue membrane. In a test tube, containing 1 ml of 3.8% sodium citrate solution, 4 ml of blood was taken, mixed and the erythrocytes were separated by centrifugation for 10 min at 3000 rpm. Plasma was removed. Transferred 1 ml of erythrocyte mass in a test tube containing 3 ml of a solution of methylene blue (0.025%), prepared in saline. The samples were stirred and incubated for 10–12 min at room temperature, then centrifuged again for 10 min at 3000 rpm. The supernatant was transferred to a cuvette and the optical density relative to saline was determined at a wavelength of 630 nm on a spectrophotometer. The amount of absorbed dye (as a percentage) was calculated from the difference between the optical density of the initial dye solution and the dye solution after incubation with erythrocytes (
The results were statistically analyzed by means of non-parametric indices methods in the STATISTICA 10.0 software (Statsoft, USA). The reliability of the differences in values between independent quantitative values was determined with a normal distribution according to the Mann-Whitney U criterion (
The level of endotoxicosis is determined by the content of hydrophilic and hydrophobic products in the blood. Middle molecular weight molecules belong to the hydrophilic components of endogenous intoxication. They are formed due to increased proteolysis and have an alternative effect on the body due to high functional activity (
As it can be seen from the table, the content of middle molecular weight molecules, which has been determined at a wavelength of 254 nm in the serum (chain amino acids), has increased on the 14th day – by 1.28 times (p < 0.01) compared with the intact group. There has also been an increase in serum middle molecular weight molecules, which has been determined at a wavelength of 280 nm (aromatic amino acids). On the 14th day, this indicator has retained this vector of direction of change, and was higher relative to the intact group of animals (by 1.46 times; p < 0.01).
Increased permeability of erythrocyte membranes is a reflection of disorders of the functions and structure of plasma membranes of all cells of the body. The erythrocyte intoxication index characterizes the inhibition of sorption activity (including methylene blue), which is a consequence of the rearrangement of lipid components of cell membranes and a decrease in the functional capacity of erythrocytes due to exposure to toxic substances (
As a result of the study of the total toxic effect on the erythrocyte membranes of the blood of experimental animals with experimental bacterial-immune periodontitis – the level of erythrocyte intoxication index (EII), significant changes were also found (Table). In particular, it was found, that on the 14th day of this experimental model of periodontitis, there has been a further gradual increase of EII (by 1.75 times; p < 0.01) compared with the intact group of animals, indicating a decrease in the adsorption capacity of erythrocytes throughout period of inflammation.
The content of middle molecular weight molecules in the serum and erythrocyte index of intoxication of experimental animals under the condition of experimental periodontitis with the use of quercetin (M ± m).
Conditions and indicators of the experiment | Control. Intact group | White rats with experimental periodontitis | |
---|---|---|---|
No correction | With quercetin correction | ||
Experiment duration (days) | - | 14 | 14 |
Number of animals | 10 | 8 | 8 |
MMWM 254, condit. units | 353.80 ± 4.53 | 454.00 ± 10.39 | 419.00 ± 10.23 |
p1 < 0.01 | p1 < 0.01; p2 < 0.01 | ||
МMWМ280, condit. units | 144.30 ± 4.67 | 210.75 ± 10.93 | 185.75 ± 7.73 |
p1 < 0.01 | p1 < 0.01; p2 < 0.05 | ||
Erythrocyte intoxication index, % | 31.06 ± 1.88 | 54.30 ± 1.66 | 49.55 ± 1.41 |
p1 < 0.01 | p1 < 0.01; p2 < 0.05 |
Analyzing the results of the study of endogenous intoxication in the serum of animals with experimental bacterial-immune periodontitis receiving quercetin, it should be noted, that there has been a significant decrease in the hydrophilic components of endogenous intoxication, namely middle molecular weight molecules, which were determined at a wavelength of 254 nm – by 1.08 times (p < 0.05), compared with data from animals with our simulated pathology on the 14th day of the experiment without the introduction of flavonol (Fig.
Dynamics of content middle molecular weight molecules (254 nm) in the blood serum of white rats with experimental periodontitis with quercetin correction (in % of control). Notes: * – significant of differences in relation to the intact animals (p < 0.01); # – # – significant of differences in relation to the animals with periodontitis on the 7th day of experiment without correction (p < 0.05).
However, when comparing the indicator MMWM254, which took place on the 14th day of development of experimental periodontitis with correction, and the indicators of the control group of animals, it was found that it remained significantly higher (by 1.18 times; p < 0.01) than animals comparison groups.
Analyzing the changes in the level of middle molecular weight molecules, which were determined on a spectrophotometer at a wavelength of 280 nm, in the blood serum of experimental animals with periodontitis which received quercetin during the development of the pathological process, it should be noted that the therapeutic efficacy of this tool is confirmed by lower serum MMWM280 (by 1.14 times, p < 0.05), compared with animals studied on the 14th day, and have not received this medicine (Fig.
Dynamics of content middle molecular weight molecules (280 nm) in the blood serum of white rats with experimental periodontitis with quercetin correction (in % of control). Notes: * – significant of differences in relation to the intact animals (p < 0.01); # – significant of differences in relation to the animals with periodontitis on the 7th day of experiment without correction (p < 0.05).
It should be noted that when comparing the above levels of aromatic amino acids on the 14th day of development of experimental periodontitis with correction, we found that they remained slightly higher than the control group of animals – by 1.29 times (p < 0.01).
According to the results of the study, it was found that there has been a significantly reduced erythrocyte intoxication index in animals treated with quercetin. The effectiveness of flavonol on the course of the inflammatory process in this simulated pathology is confirmed by a decrease of EII (by 1.10 times; p < 0.05), compared with animals with bacterial-immune periodontitis on the 14th day which did not receive the medicine (Fig.
Dynamics of erythrocyte intoxication index in the blood serum of white rats with experimental periodontitis with quercetin correction (in % of control). Notes: * – significant of differences in relation to the intact animals (p < 0.01); # – # – significant of differences in relation to the animals with periodontitis on the 7th day of experiment without correction (p < 0.05).
However, when comparing the changes in the studied index of endogenous intoxication observed on the 14th day of experimental periodontitis after quercetin injections, it was found that after correction it remained significantly higher relative to the control group of animals (by 1.60 times; p < 0.01).
One of the difficult problems of modern dentistry is periodontal disease. In recent years, along with the known concepts of the pathogenesis of inflammatory and inflammatory-dystrophic periodontal diseases, much attention has been paid to the activation of lipid peroxidation. In violation of antioxidant protection, free radical oxidation in the periodontium develops avalanche-like. The level of peroxidation of phospholipids of cell membranes increases with the destruction of the latter and the death of periodontal cells with the release of endogenous toxins. Cell division is disturbed and inert products of peroxide denaturation of lipids and proteins accumulate. It is known, that uncontrolled LPO reactions can not only lead to metabolic disorders, but also cause structural changes in tissues, suppress the body’s defense mechanisms, which in turn promotes the activation of microbes that colonize the gums and periodontal pockets. There is evidence that the activation of LPO processes is an important pathophysiological mechanism of endogenous intoxication (
Endogenous intoxication not only accompanies most diseases, but is an important factor in their pathogenesis and in many cases determines the possible adverse effects, because an important feature of MMWM is their high biological activity. They have neurotoxic activity, inhibit protein synthesis, promote hemolysis of erythrocytes, inhibit erythropoiesis and enzyme activity, cause a state of secondary immunosuppression (
The syndrome of endogenous intoxication is accompanied by increased tissue breakdown, increased catabolic processes due to the accumulation of excessive amounts of biologically active substances, deformed protein metabolites and other toxic substances of endogenous origin (
This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.