FUNCTIONING OF NO-CYCLE IN THE ORAL FLUID IN CHILDREN OF PRIMARY SCHOOL AGE WITH TYPE 1 DIABETES MELLITUS IN THE TREATMENT OF CHRONIC CATARRHAL GINGIVITIS.

OBJECTIVE: The aim: To determine the activity of NO-synthase and arginase in oral fluid in children with type 1 diabetes mellitus and to evaluate the efficacy of the treatment scheme we elaborated in the treatment of chronic catarrhal gingivitis. PATIENTS AND METHODS: Materials and methods: 82 children were examined, they were divided into groups by presence of gingivitis and diabetes mellitus. NO-synthase (NOS) activity was determined in oral fluid by the difference in nitrite concentration before and after incubation. The arginase activity was determined in oral fluid by the difference in the concentration of L-ornithine before and after incubation. RESULTS: Results: Use our treatment scheme in children with chronic catarrhal gingivitis and type 1 diabetes mellitus lead to a change in the polarization of oral macrophages towards the predominance of M2 polarization in 1 month. The polarization of macrophages changed to the predominance of M1 polarization activity in 1 year. CONCLUSION: Conclusions: We have elaborated a scheme for the treatment of chronic catarrhal gingivitis in children with type 1 diabetes mellitus. It normalizes the polarization of oral macrophages caused by exposure to chronic catarrhal gingivitis as a local pathogenetic factor.

BIOFLAVONOIDS AS AGENTS FOR CORRECTING NITRO-OXIDATIVE STRESS AND SALIVARY GLAND FUNCTIONS IN RATS EXPOSED TO ALCOHOL DURING MODELED LIPOPOLYSACCHARIDE-INDUCED SYSTEMIC INFLAMMATORY RESPONSE.

OBJECTIVE: The aim: To investigate the effects of bioflavonoids (curcumin, epigallocatechin-3-gallate and quercetin) on nitro-oxidative stress and the functions of submandibular SGs in rats under alcohol exposure during SIR. PATIENTS AND METHODS: Materials and methods: The studies were conducted on 35 rats of the Wistar line weighing 205-220 g, divided into 5 groups of seven animals in each: the 1st group, control group I, included animals receiving isotonic sodium chloride solution intragastrically twice a day; the 2nd group, control group II, included rats exposed to alcohol (in a dose of 24 mg/kg intragastrically through gavage a twice a day) for last 2 weeks during lipopolysaccharide (LPS)-induced SIR; the rats of the 3rd, 4th and 5th groups exposed to alcohol during LPS-induced SIR, which also received bioflavonoids. The bioflavonoids ("Sigma-Aldrich, Inc.", USA) were as following: curcumin (in a daily dose of 200 mg/kg), epigallocatechin-3-gallate (in a daily dose of 40 mg/kg), and quercetin (in a daily dose of 200 mg/kg), respectively. SIR was induced by intraperitoneal administration of Salmonella typhi LPS (during the first week a dose of 0.4 µg/kg of body weight was administered 3 times a week; during the next 7 weeks of the experiment rats received 0.4 µg/kg of body weight once a week. The formation of superoxide anion radical (О2 -), activity of NO-synthase - total (NOS), its constitutive and inducible isoforms (cNOS, iNOS), and concentration of peroxynitrites and S-nitrosothiols were evaluated spectrophotometrically. To assess the functional status of submandibular SGs in their homogenate we determined α-amylase activity (spectrophotometrically) and the aquaporin-5 concentration (by enzyme-linked immunosorbent assay). through gav-age with orogastric cannul. RESULTS: Results: When applying bioflavonoids under the conditions of alcohol administration during SIR, NADH-induced .О2 - production decreased and yielded to the result in the control group II by 36.8% under administering curcumin, by 34.5% under administering epigallocatechin-3-gallate, and by 41.3% under administering quercetin. The total NOS activity in SGs tissues was inferior by 42.8% to the relevant data in the control group II (under curcumin administration), by 33.7% (under epigallocatechin-3-gallate administration) and by 46.6% (under quercetin administration); and the iNOS activity decreased by 47.0, 38.3 and 52.0%, respectively. Under the administration of bioflavonoids peroxynitrites concentration in the submandibular SGs tissues was inferior to the control group II by 35.6% (under curcumin administration), by 37.4% (under epigallocatechin-3-gallate administration), and by 39.3% (under quercetin administration); the content of S-nitrosothiols was lower by 34.5, 31.1 and 35.3%, respectively. The administration of bioflavonoids led to the changes in α-amylase activity in the submandibular SGs tissues: its values exceeded the relevant data in the control group II by 40.4% (under curcumin administration), by 38.2% (under epigallocatechin-3-gallate administration), and by 34.1% (under quercetin administration); under those conditions aquaporin-5 concentration grew in 2.66, 2.61 and 2.55 times, respectively. CONCLUSION: Conclusions: The use of bioflavonoids (curcumin, epigallocatechin-3-gallate, and quercetin) under the combined administration of 40% ethanol solution and LPS considerably limits the development of nitro-oxidative stress in the tissues of the submandibular SGs. The administration of the bioflavonoids increases the level of cNOS coupling, and improves the functional status of the submandibular SGs under the combined administration of alcohol and LPS enhancing the activity of α-amylase and concentration of aquaporin-5.

QUERCETIN LIMITS THE PROGRESSION OF OXIDATIVE AND NITROSATIVE STRESS IN THE RATS' TISSUES AFTER EXPERIMENTAL TRAUMATIC BRAIN INJURY.

OBJECTIVE: The aim: To investigate the effect of water-soluble form of quercetin on the indices reflecting the progression of oxidative-nitrosative stress in the cerebral tissues and the periodontium of rats after experimental TBI. PATIENTS AND METHODS: Materials and methods: The studies were conducted on 30 white rats of the Wistar line weighing 180-220 g, divided into 3 groups: the 1st group included pseudo-injured animals (subjected to ether anaesthesia, fixation without TBI modeling), the 2nd group included the animals exposed to modeled moderate TBI, the 3rd group involved the rats, which were given injections with water-soluble form of quercetin (corvitin, "Borshchahivskiy CPP", Ukraine) intraperitoneally in a daily dose of 10 mg/kg recalculated for quercetin for 7 days following the TBI modeling. The formation of superoxide radical anion (.О2 -), activity of NO-synthase - total (NOS), its constitutive and inducible isoforms (cNOS, iNOS) - and concentration of peroxynitrite were evaluated spectrophotometrically. The level of lipid peroxidation (LPO) in the tissues was evaluated by the formation of a stained trimethine complex during the reaction of tiobarbituric acid (TBA). The activity of the antioxidant system was assessed by increasing in the concentration of TBA active products during 1.5 hour incubation in iron-ascorbate buffer solution, as well as by the activity of antioxidant enzymes - superoxide dismutase (SOD) and catalase. RESULTS: Results: The use of quercetin under the experimental conditions significantly reduced the О2 - generation by NADPH- and NADH-dependent electron transport chains by 30.2 and 35.0% (in the cerebral hemispheres) and by 23.5 and 32.5% (in the soft periodontal tissues), respectively, compared to the findings in the 2nd group. The production of this radical by leukocyte NADPH oxidase in these organs was inferior to the value of the 2nd group by 39.3 and 29.9%. We revealed that the use of quercetin in the experimental conditions probably reduced the activity of NOS, including iNOS, by 38.2 and 45.3% (in the cerebral hemispheres) and by 53.5 and 66.9% (in the soft periodontal tissues), respectively, compared with the findings in the 2nd group. Under these conditions, the cNOS activity went up by 50.0% and doubled, the peroxynitrite content was lower by 19.5 and 32.1% than that in the 2nd group. The administration of quercetin in the experimental conditions significantly reduced the concentration of TBA-active products in the homogenate of cerebral hemispheres and soft periodontal tissues. The development of decompensated LPO is also confirmed by a decrease in the activity of SOD and catalase. CONCLUSION: Conclusions: on the 7th day after modeling moderate TBI in rats the signs of oxidative-nitrosative stress are found not only in locus morbi (in the tissue of the cerebral hemisphere), but also in distant organs (periodontal tissues). Applying of water-soluble form of quercetin significantly reduces signs of oxidative-nitrosative stress in the tissue of the cerebral hemisphere of rats, as well as in periodontal tissues on the 7th day after moderate TBI modeling.

Quercetin potentiates antiradical properties of epigallocatechin-3-gallate in periodontium of rats under systemic and local administration of lipopolisaccharide of salmonella typhi

Introduction: There has been demonstrated that pharmaceutical effect of epigallocatechin-3-gallate (EGCG), a polyphenol, which is found in green tea (Camellia sinensis), is implemented through the activation of Nrf2 (Nuclear Factor Erythroid 2-Related Factor 2).The importance of Keap1 / Nrf2 / antioxidant response element (ARE) system is determined by the fact that the state of NF-κB- and АР-1-associated pathways depends on its activity. Recent studies have demonstrated the property of quercetin to suppress ubiquitin-dependent proteolysis of complex of NF-κB and its inhibitory protein IκB. All this provides preconditions to eliminate the potentiality of NF-κB-dependent expression of the number of genes of pro-oxidant and pro-inflammatory proteins. However, co-effect produced by quercetin and EGCG on the oxidative nitrosative stress markers in the periodontal tissues is still unclear. The aim: To investigate the co-effect produced by quercetin and an inducer of the Keap1 / Nrf2 / ARE epigallocatechin-3-gallate on markers of oxidative-nitrosative stress in rats' periodontium under the systemic and local administration of Salmonella typhi lipopolysaccharide (LPS). Material and methods: The studies were conducted on 30 white rats of the Wistar line, divided into 5 groups: the 1st included intact animals, the 2nd was made up of animals after their exposure to combined systemic and local LPS administration, the 3rd and 4th groups included animals, which were given injections with water-soluble form of quercetin (corvitin) and EGCG respectively, and the 5th group involved rats, which were injected with co-administered corvitin and EGCG. The formation of superoxide anion radical (.О-2 ) was evaluated by a test with nitro blue tetrazolium using spectrophotometry of the periodontal soft tissue homogenate. The total activity of NO-synthase and concentration of peroxynitrite in the homogenate of the soft components of periodontium were evaluated spectrophotometrically. Results: Co-effect produced by corvitin and EGCG under systemic and local LPS administration is accompanied with reduced О-2 production by NADPH-dependent electron transport chains (microsomal and NOS) by 20.0 % (p <0.05) compared with values for the animals received separate corvitin during the experiment. .О-2 generation by the mitochondrial respiratory chain yielded to comparable data of the 3rd and 4th groups by 27.6 % (p <0.01) and 23.8 % (p <0.05) respectively. No differences were found between the groups exposed to combined or separate action of the above mentioned agents in the experiment when assessing О-2 generation by leukocyte NADPH-oxidase. Combined effect of corvitin and EGCG during systemic and local LSP administration showed the decrease in NOS activity and peroxynitrite concentration in periodontal tissues by 53.3 % (p <0.001) and 27.0 % (p <0.02) compared with the findings in the 3rd group, and by 42.0 % (p <0.01) and 22.3 % (p <0.01) in the 4th group. Conclusions: The co-administration of water-soluble form of quercetin and epigallocatechin-3-gallate under systemic and local introducing of lipopolysaccharide Salmonella typhi has been proven to be more effective means for preventing and correcting oxidative-nitrosative stress in the periodontal tissues than this occurs at separate administration of each of the polyphenols.

Epigallocatechin-3-gallate prevents disruption of connective tissue in periodontium and salivary glands of rats during systemic inflammation.

OBJECTIVE: Introduction: The connective tissue remodeling is essential for periodontal and salivary glands (SG) pathology. Recently there has been demonstrated the number of pharmacological effects of green tea (Camellia sinensis) such as antioxidant, anti-inflammatory, anti-aging, antibacterial, antiviral and DNA-protective effects, associated with the presence of epigallocatechin-3-gallate (EGCG) as an inducer of the Keap1 / Nrf2 / antioxidant response element signaling pathway. However, the EGCG effects on the components of soft connective tissues of periodontium and SG are still unclear. The aim: To investigate the effect of EGCG on markers of disruption of periodontal and submandibular SG connective tissues in rats during the conditions of experimental systemic inflammation (SI). PATIENTS AND METHODS: Materials and methods: The studies were conducted on 30 white rats of the Wistar line, divided into 3 groups: the 1st included intact animals, the 2nd was made up of animals after induced SI (by intraperitoneal administration of lipopolysaccharide Salmonella typhi), and the 3rd included animals, which were injected EGCG (production of Sigma-Aldrich, Inc., USA) intraperitoneally in a dose of 21.1 mg / kg 3 times a week, starting on the 30th day of SI induction. The level of collagenolysis was assessed by the content of free hydroxyproline (FHP). The process of depolymerization of proteoglycans and sialoglycoproteins was evaluated by determining their monomers, glycosaminoglycans (GAGs) and N-acetylneuraminic acid (NANA) respectively. The molar roots exposure index (MREI) was calculated. RESULTS: Results: Administering EGCG reduced the content of FHP by 33.3 % (p<0.01), the content of GAGs by 39.4% (p<0.02), and content of NANA by 34.3% (p<0.001) in the soft periodontal tissues compared with the relevant findings in the second group of the animals. In this condition the concentration of these compounds in the calcified components of periodontium (alveolar bone) lowered as well: FHP - by 41.9% (p<0.001), GAGs - by 41.0% (p<0.001), NANA - by 53.3% (p<0.001), MREI reduced to 27.1+1.6, i.e. by 27.7% (p<0.01) compared with the relevant findings in the second group of the animals. The administration of EGCG also reduced the content of FHP by 37.8% (p<0.001), the content of GAGs by 39.8% (p<0.001), and the content of NANA by 37.6% (p<0.001) in SG tissues compared with the relevant results of the second group of the animals. CONCLUSION: Conclusions: The administration of EGCG under modeled systemic inflammation is an effective means of preventing and correcting the disruption of connective tissue of periodontium and submandibular salivary glands in rats: it reduces collagenolysis and depolymerization of proteoglycans and glycoproteins.