[FULLERENE C60 INHIBITED FREE RADICAL AND DESTRUCTIVE PROCESSES IN CONNECTIVE TISSUE DURING ADJUVANT ARTHRITIS IN RATS].

The effects of fullerene C60 (FC60) on the level of free radical and destruction processes were studied in rats with experimental adjuvant arthritis (AA). It was shown the protective effect of FC60 during AA. The effect was accompanied by an increase of the antioxidant enzymes activity, superoxide dismutase in the liver (15.96 ± 0.38 µmol/kg x s) and in the kidneys (5.36 ± 0.27 µmol/kg x s) and catalase in the kidneys (9.56 ± 0.78 µmol/kg x s) and in the heart (2.26 ± 0.41 µmol/kg x s) in comparison to control group (43.83± 5.69%; 54.55 ± 6.18%; 11.68 ± 0.52 µmol/kg x s; 3.43 ± 0.47 µmol/kg x s; 4.77 ± 0.5 µmol/kg x s; 0.98 ± 0.12 µmol/kg x s accordingly). It was shown a protective effect of FC60 during AA directed on the depression of the destructive processes in connective tissue that was expressed through the reduction of the total collagenolitic activity level in cartilage (10.05 ± 0.06 µmol/g/min) and bone (11.21 ± 0.04 µmol/g/min) tissues, free hydroxyproline contents (1.54 ± 0.04 µg/ml) and alkaline phasphatase activity (1.24 ± 0,14 µmol/l x sec) in comparison to control group (11.91 ± 0.49 µmol/g/min; 13.19 ± 0.15 µmol/g/min; 2.25 ± 0.07 µg/ ml; 2.19 ± 0.24 µmol/l x sec accordingly). Taken together, these results accentuate the perspective of future investigations of action FC60 during rheumatoid arthritis as a feasible therapeutic agent.

[The anti-inflammatory effect of fullerene C60 on adjuvant arthritis in rats].

The anti-inflammatory properties of non-modified fullerene C60 (FC60) by adjuvant arthritis in Wistar rats have been studied. It was shown that the intraperitoneal introduction of FC60 (50 ng) reveals an anti-inflammatory and chondroprotective actions in the phase of systemic manifestation of adjuvant arthritis. The effect was carried out by limitation of inflammation of damaged limb, normalization of body weight, the decrease in body temperature. Introduction of FC60 promote the reduction of leukocyte level, the erythrocyte sedimentation rate, concentrations of sialic acids and the ceruloplasmin levels, processes degeneration of cartilaginous tissues of the joint of rats. It has been concluded that the therapeutic effectiveness of non-modified FC60 in experimental adjuvant arthritis is comparable with the action of water-soluble forms of fullerenes. The results substantiate the future investigations of non-modified FC60 for design of therapeutic agents for treatment of rheumatoid arthritis in clinics.

[The condition of lipid peroxidation in mice and the effect of fullerene C60 during immune response].

The aim of this study was to assess the influence of fullerene C60 on lipid peroxidation (POL) and antioxidant protection during the induction of the immune response to heteroantigen. Balb/c mice were immunized intraperitoneal (i.p.) with sheep erythrocytes for the primary immunization. Water dispersion of fullerene C60 was injected i.p. once at the dose 50 ng to mice on first, third and sixth days after immunization. During immune response, the increment ofmalonic dialdehide (MDA) was enhanced in liver, kidneys and heart tissues. Fullerene C60 induced POL during the latent phase of immune response, but inhibited this process during progression of immune response. Activities of superoxide dismutase (SOD) and catalase in liver and spleen tissues were induced after injection of fullerene C60 to intact mice. After immunization, high level of activity of antioxidant enzymes and low level of organs mass factor were determined. Injection of fullerene C60 reduced the activities of SOD and catalase in spleen tissues. The results of our study indicate that fullerene C60 can display positive effect on POL processes and antioxidant enzymes activity which is probably due to membrane's stabilization action or the ability of fullerene C60 to bind free radicals independently.

[Fullerene C60 exhibits immunomodulatory activity during adjuvant-induced arthritis in rats].

The effect of fullerene C60 (FC60) on the immune processes during experimental adjuvant-induced arthritis (AA) in rats has been studied. The results indicate the inhibitory action of FN60 during AA on cellular splenocyte proliferation, neutrophil phagocytic and oxygen-stimulatory activities in the NBT test, and humoral immune mechanisms involved in the production of antinuclear antibodies, formation of circulating immune complexes, and restoration of morphological structure of spleen. Taken together, these results allow FC60 to be considered as a new potential pharmacological agent that can realize its effects mainly through anti-inflammatory and immunomodulatory activity.

[Pioglitazone, an activator of PPAR-gamma, reduces the expression of kB nuclear factor and inhibits apoptosis in mononuclear cells of peripheral blood in vitro].

In order to examine the impact of PPAR-gamma activator on the expression of kB nuclear transcription factor and apoptosis, the peripheral blood mononuclear cells (PBMC) were incubated with pioglitazone at concentrations 10, 30 and 100 MM. Expression of NF-kB in CD64+ cells was evaluated by flow cytometry using monoclonal antibodies. The development of apoptosis was evaluated using annexin V and propidium iodide. Activation of PPAR-y resulted in a reduction of the number of CD64+ cells and expression of the maximum levels of these receptors. We also obesrved a dose-dependent decrease of NF-kB concentration in PBMC suspension and in CD64+ cells. While analyzing the apoptosis, the reduction of apoptotic cells (reduction of AnV+PI+ cells) has been observed. The results obtained allow to prove that PPAR-gamma activation participates in the PBMC apoptosis regulation and in the expression of transcriptional factor NF-kB. Our results indicate for the involvement of PPAR-gamma in the regulation of NF-kB activity which plays one of the most significant roles in the inflammation realization, oncogenesis, and so on.

[Quantitative analysis of individual groups of microorganisms, extracted from atherosclerotic lesions in the coronary arteries in patients depending upon ASP299GLY polymorphism of TLR4 gene].

The estimation data of contamination by separate groups of microorganisms and dependence of the microbial content level upon TLR4 gene 896A/G polymorphism in 20 samples of atherosclerotic lesions in coronary arteries has been presented. The presence of TLR4 gene polymorphic allele G in the individual genotype determines the increased contamination of atherosclerotic plaque tissues by the representatives of the following genera: Lactobacillus sp., Enterobacterium sp., Sneathia sp./Leptotrihia sp./Fusobacterium sp., Mobiluncus sp./Corynebacterium sp., Peptostreptococcus sp. The emergence of new correlation pairs with participation of Lachnobacterium sp./Corynebacterium sp. among the carriers of G allele has been revealed via the intragroup correlation analysis. The obtained results confirm the possible involvement of the represented groups of microorganisms in the pathogenesis of atherosclerosis and the role of the TLR4 gene polymorphic variant G in the increased microbial contamination of the coronary arteries tissues.

[Relationship of toll-like receptors 2 and 4 genes polymorphisms with allergic diseases with increased levels of specific IgE].

The aim of the research was to identify the role of genes polymorphisms encoding TLR2 (NP_003255.2) and TLR4 (NP_612564.1) in the increase of the specific IgE production level and development of allergic diseases. Genetic typing of genome specific regions was performed by polymerase chain reaction method using specific oligonucleotide primers. The obtained results display that the most significant causative allergens are epidermal and household allergens (epidermis of cat, dog and horse, D. farinae, D. pteronyssinus). The relationship of polymorphisms TLR2 (rs5743708) and TLR4 (rs4986790) with increased production of specific IgE in patients with allergic diseases was also observed.

[Effect of fullerene C60 on functional activity of phagocytic cells].

The effect of aqueous dispersion of fullerene C60 (FC60) on the functional activity of cells involved in the phagocytosis reactions was studied. FC60 (0.01 microM/l and 0.1 microM/l) produced mainly negative effects on the activity ofnonspecific immunity cells by inhibiting the myeloperoxidase enzymatic activity, decreased the level of induced chemiluminescence, and suppressed the expression of molecules CD54 involved in the adhesion. The only exception was a slight stimulating effect on the NBT test. The results indicate the FC60 influences various stages and mechanisms of phagocytosis.

[Effect of kidney peptide preparation on the expression of lymphocyte surface receptors during Na+,K+-ATPase inhibition]. / Vplyv peptydnoho preparatu z nyrok na riven' ekspresiï poverkhnevykh retseptoriv limfotsytiv za umov pryhnichennia aktyvnosti Na+,K+-ATFazy.

In the work the action of membrane-mediated kidney peptide complex on human periferal blood lymphocytes was studied under Na+,K+-ATPase activity inhibition. It was shown that ouabain pretreatment changed the functional activity of lymphocytes: the surface immunoglobulin expression level was decreased, antigene determinants CD4, CD8 expression were reduced, CD3 expression was increased either. The kidney peptide complex had the modulated influence on the surface receptors, improving the decreased CD4, CD8 and surface immunoglobulins expression and differently influencing the CD3 and CD72 expression. It has been proposed that kidney peptide complex facilitates processes of Na+,K+-ATPase reactivation.

[The role of calcium-dependent mechanism in the realization of immune effects of kidney peptide complex]. / Rol' kal'tsii zalezhnykh mekhanizmiv u realizatsiï imunotropnykh efektiv peptydnoho kompleksu nyrok.

The role of intracellular and extracellular calcium in the realization of immune effects of the kidney peptide complex were studied. Expression of surface immunoglobulin receptors on lymphocytes depressed under the influence of calcium channels blockators (verapamil, phenigidin), binding of intracellular and extracellular calcium (EDTA, BAPTA). The kidney peptide complex restorted expression of surface immunoglobulins depressed by verapamil, phenigidin, EDTA, BAPTA. Also in this conditions the peptide complex increased the re-assembly of receptors, as caps, patches and clasters. Kidney peptide complex has a synergy with the calcium ionophor A 23187 to increase the expression of surface immunoglobulin receptor on lymphocytes. We could not exclude the direct influence of peptide complex on the conformation changes of the ion traffic systems for calcium. It was suggested that peptide complex promote the entrance of calcium ions into the cells. The influence of kidney peptide complex on expression of lymphocytes membrane receptors are independent from the calcium existence neither into cells nor outside the cells.

[Effect of paradentium polypeptide on the lipid peroxidation of the membranes and coagulation of erythrocytes in rats with spontaneous paradentitis]. / Vliianie polipeptida parodonta na perekisnoe okislenie lipidov éritrotsitarnykh membran i koaguliatsionnuiu aktivnost Eritrotsitov u krys pri spontannom parodontite.

The influence of alkaline polypeptides-cytomedines on the level of free radical oxidation of lipids, hemocoagulant, fibrinolytic and aggregative properties of erythrocytes in spontaneous parodontitis has been investigated. It has been established that the development of spontaneous parodontitis is accompanied by the activation of peroxide oxidation of cellular membranes, which results in the development of hypercoagulation and inhibition of fibrinolytic properties of erythrocytes, thereby intensifying the state of microcirculatory hemostasis. Polypeptide of the parodontium a modulating effect on the state of free-radical oxidation, improves antioxidant status of blood cells, stabilizes indices of erythrocytic link of hemostasis. The investigated cytomedines may be promising for treatment of the parodontium pathology.