CHANGES IN BIOCHEMICAL PARAMETERS OF BLOOD SERUM AND GINGIVAL HOMOGENATES WITH EXPERIMENTAL GINGIVITIS

the NO-synthase system and the development of inflammatory processes in rats of different ages in the dynamics of tobacco smoke poisoning. Studies were carried out on white, non-native male rats of different ages, which were exposed to tobacco smoke daily for 45 days in a sealed chamber (exposure time 6 min). After 15, 30 and 45 days from the onset of the animals' defeat with tobacco smoke, they were taken out of the experiment by euthanizing them under thiopental anesthesia, observing all the rules for working with vertebrates. It was found that the damage to the rats for 45 days with tobacco smoke led to an increase in the serum content of the proinflammatory cytokine IL-6. The most sensitive to this toxicant were immature rats, in which the content of IL-6 increased by 50%, 164% and 237%, respectively, 15, 30 and 45 days after injury. Poisoning of rats with tobacco smoke led to an increase in the activity of inducible NO-synthase in serum and liver throughout the study period. It is noted that the body of immature rats reacts most quickly and most strongly to tobacco smoke. The activity of iNO synthase reached the highest values both in the blood serum and in the liver of immature animals. After the defeat of rats with tobacco smoke, in our experiments a decrease in the activity of endothelial NO synthase in all age groups of rats was noted. In immature rats, the activity of eNOS in serum decreased 1.4 times, 1.8 and 2 times in 15, 30 and 45 days, respectively, compared with the norm. In the liver of rats of all age groups, during the experiment, a similar decrease in eNOS activity was observed and the lowest value was reached at the last term of the study (day 45 of the exposure to tobacco smoke). Inflammatory periodontal diseases is an urgent problem not only in dentistry, but also in general medicine, due to their high prevalence and negative effects on a human body causing significant economic consequences. As a result, the bioregulation of biochemical processes is often damaged, due to it, at a certain level of damage, different diseases, including dental ones, occur and develop. Moreover, the study of the impact of the combined action of xenobiotics on biochemical tissue indicators of a growing up human body, the development of new means of correction, which are based on inhibition of proteases, hydrolases, kinin system, normalization of tissue penetration, enhancing the action of protective factors of a human body, ensuring the sustainability of periodontal tissues to the negative influence of environmental agents, are important and poorly understood issues. Research purpose – to study the biochemical parameters of blood serum and homogenates of the gums in experimental animals with gingivitis caused by the combined action of heavy metals and deficiency of fluorine and iodine. Consequently, experimental studies have convincingly demonstrated that in experimental gingivitis under the influence of the adverse environmental factors the levels of inflammatory markers ‒ elastase and urease, as well as the indices of the protease-inhibitory system increased.

the NO-synthase system and the development of inflammatory processes in rats of different ages in the dynamics of tobacco smoke poisoning. Studies were carried out on white, non-native male rats of different ages, which were exposed to tobacco smoke daily for 45 days in a sealed chamber (exposure time 6 min). After 15, 30 and 45 days from the onset of the animals' defeat with tobacco smoke, they were taken out of the experiment by euthanizing them under thiopental anesthesia, observing all the rules for working with vertebrates. It was found that the damage to the rats for 45 days with tobacco smoke led to an increase in the serum content of the proinflammatory cytokine IL-6. The most sensitive to this toxicant were immature rats, in which the content of IL-6 increased by 50%, 164% and 237%, respectively, 15, 30 and 45 days after injury. Poisoning of rats with tobacco smoke led to an increase in the activity of inducible NO-synthase in serum and liver throughout the study period. It is noted that the body of immature rats reacts most quickly and most strongly to tobacco smoke. The activity of iNO synthase reached the highest values both in the blood serum and in the liver of immature animals. After the defeat of rats with tobacco smoke, in our experiments a decrease in the activity of endothelial NO synthase in all age groups of rats was noted. In immature rats, the activity of eNOS in serum decreased 1.4 times, 1.8 and 2 times in 15, 30 and 45 days, respectively, compared with the norm. In the liver of rats of all age groups, during the experiment, a similar decrease in eNOS activity was observed and the lowest value was reached at the last term of the study (day 45 of the exposure to tobacco smoke).
Inflammatory periodontal diseases is an urgent problem not only in dentistry, but also in general medicine, due to their high prevalence and negative impact on a human body causing considerable economic consequences [1]. For the last ten years state expenses for providing dental care to children of Ukraine have increased more than 2.5 times, indicating the lack of effectiveness of the prevention of major dental diseases in children [2,3]. The scientific literature extensively discusses the influence of ecological factors of the environment on the children's morbidity, including a dental one [4,5]. It is known, that due to anthropogenic pollution of the biosphere, water and soil up to 70% of toxicants get into a human body, and under the deficiency of certain microelements (iodine, fluorine, selenium, iron) cause pathogenic polytropic changes in a child's body [2,6]. Because of the high prevalence of toxicants in the environment, a particular interest is aroused to diseases caused by their action, which can manifest themselves at the molecular, cellular, tissue and body levels [7]. As a consequence, the bioregulation of biochemical processes is often damaged, due to it, at a certain level of damage, different diseases, including dental ones, occur and develop. Therefore, the study of the impact of the combined action of xenobiotics on biochemical tissue indicators of a growing up human body, the development of new means of correction, which are based on inhibition of proteases, hydrolases, kinin system, normalization of tissue penetration, enhancing the action of protective factors of a human body, ensuring the sustainability of periodontal tissues to the negative influence of environmental agents, are important and poorly understood issues.
Research purposeto study the biochemical parameters of blood serum and homogenates of the gums in experimental animals with gingivitis caused by the combined action of heavy metals and deficiency of fluorine and iodine.
Мaterial and methods. Experimental gingivitis in rats, aged 30±5 days, was induced by means of the peroxide gingivitis model, by adding to their normal diet peroxide sunflower oil at a dose of 1 ml per animal within 3 weeks [8]. Eighty white rats with average weight 54±5 g, males and females equally, were included in the study. Depending on the design of anthropogenic environmental conditions the rats were assigned to four groups: I group (control) -20 intact rats, which were held on a regular vivarium diet; II group -20 animals with simulated gingivitis; III group -20 animals with induced gingivitis by means of the peroxide gingivitis model with the addition to water of heavy metals salts based on their molecular weight (CdCl2 = 0,010 mg / l; Pb (NO3) 2 = 0.36 mg / l) [9]; IV group -20 animals with induced gingivitis by means of the peroxide gingivitis model with the addition of heavy metals salts to water + iodine deficiency + fluorine deficiency. Iodine deficiency in rats was induced by adding merkazolin to water, at the dose of 50 mg / kg of body weight per day for 3 weeks. [10]. Deficiency of fluorine was simulated by keeping the animals on a low calorie diet with the exception of products containing much fluorine.
After 21 days the animals were led out of the experiment under ether anesthesia. Blood and gums samples were taken for investigation. Trypsin-inhibitory and total proteolytic activities in blood serum were measured. The total proteolytic activity (TPA) was determined by reaction with Folin's reagent The study was conducted in compliance with the general rules and regulations of the -European Convention for the Protection of Vertebrate Animals used for Experimental and other Scientific Purposes‖ (Strasbourg, 1986) and -General Ethical Principles of Animal Experiments‖ (Kyiv, 2001).
Results and its discussion. Analysis of elastase and urease activity in gum homogenates of experimental animals ( Fig. 1) showed, that in the experimental group I (intact animals) elastase activity was 0.53 ± 0.14 nKAT/ g, which was 2.3 times significantly higher than the results of the experimental group II (1.24 ± 0.19 nKAT / g, p <0.01).
The further increase of elastase activity was observed in research groups III and IV (respectively 1.53 ± 0.17 nKAT / g, and 3.22 ± 0.14 nKAT / g) which exceeded the data obtained in the group of intact animals by 3-6 times (p <0.01). It should be noted, that the values of elastase activity in the research groups III and IV, where animals were exposed to heavy metals and their combination with fluorine and iodine deficiencies on the background of gingivitis, were 1.2 and 2.6 times higher in comparison with the values of the experimental group II (p1 <0.01), where the animals had simulated gingivitis. The activity of urease in gum homogenates was 0.04 ± 0.003 nKAT / l in the group of intact animals which was by 80.0% lower than the value in the group of animals with simulated gingivitis (p <0.01). The urease activity levels in groups III and IV, where animals with gingivitis were affected by the negative factors of environment, exceeded the data obtained in the group of intact animals at respectively 130.0% and 205.0% (p <0.01). At the same time, the activity of urease in groups III and IV was by 27.8% and 69.4% higher (p1 <0.01) in comparison with the relevant data in group II.
Evaluation of biochemical parameters -the total proteolytic activity (TPA), trypsin inhibitor activity (TIA), and their ratio (TIA / TPA), in the blood serum of experimental animals (Fig. 2), showed that PAD level in group II (animals with simulated gingivitis) was significantly (p <0.01) higher by 12.40% in comparison with the value in intact animals (group I).