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Using the model of cyclophosphamide (CP)-induced immunosuppression in C57BL/6 mice, the hepatotropic effects of a conjugate of betulonic acid with 9-(4-methylpiperazin-1-ylmethyl)-2-(1,2,3-triazolyl) oreozelone (BABC) have been studied. In the liver of treated animals the expression of genes for cytochromes (CYP 1A1, CYP 1A2, CYP 3A44, CYP 2B10, CYP 2C29, CYP 17A1), PPARA, and cytokines (TNF-α, IL-1β, IL-12α, IL-10) and the relative levels of NF-κB p65, GST-π, and NAT-1 proteins were determined. On day six after administration of the compound and CP to animals a significant (3.2-fold) increase in the expression of the CYP 2B10 as compared to the control group was observed. Treatment of mice with the compound and CP also caused a 2.4-fold increase in the mRNA level of the pro-inflammatory TNF-α gene as compared to the group of animals receiving CP. Administration of the studied compound to intact animals was accompanied by a 2.5-fold increase in the IL-1β expression and a 1.8-fold decrease in the IL-10 expression as compared to the control group. An increase in the expression of pro-inflammatory cytokine genes in the liver of animals treated with the compound was accompanied by an increase in the content of NF-κB p65 (by 1.6 times), as well as an increase in the relative amount of NAT-1 protein (by 2.7 times) as compared to control animals.

The immunomodulatory activity of a betulonic acid-based compound with furocoumarin (BABCF; 2-azido, 9-N-methylpiperazinomethyl oreozelone) has been investigated. Male C57BL/6 mice (aged 3 months) treated with the cytostatic agent cyclophosphamide (CP) and intact individuals served as experimental models. The expression of genes was studied in bone marrow (IL-12, IL-10, IL-1β, TNF-α, TGF-β, M-CSF, GM-CSF) or in the suspension of peritoneal cells (IL-12, IL-10; as the injection site). The surface markers of T-lymphocytes (CD3, CD4, and CD8) in fractions of venous blood mononuclear cells (MNCs) were determined by means of flow cytometry using antibodies. Histological and morphometric studies were performed to assess the impact of CP and BABCF on the thymus. BABCF caused a pronounced (about 3-fold) increase in relative expression of the GM-KSF gene. BABCF caused a local increase in the expression of IL-12 in the peritoneal cavity cells and restored the relative content of T-lymphocytes in the blood of CP-treated mice treated affecting mainly CD3⁺CD4⁺ lymphocytes. This substance reduced the tissue density of the thymic cortex and thymic medulla in CP-treated mice. Thus, results of this study suggest that BABCF exhibits a stimulating effect on the cellular link of immunity and promotes maintenance of the number of T-lymphocytes in the blood due to their migration from the central organs of the immune system.

Chronic stress is a risk factor for major depression. Social defeat stress is a well-validated murine model of depression. However, little is known about the gene activity dynamics during the development of a depression-like state. We analyzed the effects of social defeat stress of varying duration (10 and 30 days) on the behavioral patterns and prefrontal-cortex transcriptome of C57BL/6 mice. The 10-day exposure to social defeat stress resulted in a high level of social avoidance with no signs of depression-associated behavior. Most animals exposed to 30 days of social defeat stress demonstrated clear hallmarks of depression, including a higher level of social avoidance, increased immobility in the forced swimming test, and anhedonic behavior. The monitoring of transcriptome changes revealed widespread alterations in gene expression on the 10th day. Surprisingly, the expression of only a few genes were affected by the 30th day of stress, apparently due to a reversal of the majority of the early stress-induced changes to the original basal state. Moreover, we have found that glucocorticoid-sensitive genes are clearly stimulated targets on the 10th day of stress, but these genes stop responding to the elevated corticosterone level by the 30th day of stress. The majority of genes altered by the 30-day stress were downregulated, with the most relevant ones participating in chromatin modifications and neuroplasticity (e.g., guanine nucleotide exchange factors of the Rho-family of GTPases). Very different molecular responses occur during short-term and long-term social stress in mice. The early-stress response is associated with social avoidance and with upregulation and downregulation of many genes, including those related to signal transduction and cell adhesion pathways. Downregulation of a few genes, in particular, genes for histone-modifying methyltransferases, is a signature response to prolonged stress that induces symptoms of depression. Altogether, our data show that the development of depression under social stress conditions is correlated with suppression of the overactive molecular response to induced stress, involving gene regulatory resistance to glucocorticoid molecules, potentially via a chromatin remodeling mechanism.

Interstrain differences in behavior and parameters of the immune system of CBA and C57BL/6 mice with round the clock coverage (KO) were investigated. Open field, light/dark, acoustic startle response, forced swimming, elevated plus-maze, passive avoidance were used for measuring emotional behavior and memory. The number of lymphocyte subpopulations CD3+, CD4+8-, CD4-8+, CD4+8+, CD19+, CD3+hi spleen and thymus, the ratio of cells in different phases of the cell cycle was determined by flow cytometry. C57BL/6J mice strictly increased anxiety in response to the KO compared to CBA mice. Moreover, KO-treated C57BL/6J mice impaired the passive avoidance learning. We found that KO evoked significant changes in the cellular composition of the thymus and decrease of thymocytes proliferation in C57BL/6J mice. In opposite KO-treated CBA mice showed change of splenic cellular structure with increased % CD19+ cells and the proliferation of splenocytes. Our study demonstrated genotype-dependent reactions of the nervous and immune systems in response chronic constant light.

In this work the correlation analysis was applied to detect the integrated response of the pineal gland (PG) and immunocompetent organs of male Wistar rats in response to administration of melatonin (MT) in light regime disturbances. Animals were kept for 14 days under natural or continuous light (CL). Then for 7 days they received the injections of either 0.9% solution of sodium chloride or MT, after which the rats were decapitated and the mass of their body, PG, thymus and spleen was determined. The lymphocyte subpopulations of the thymus and spleen were studied by flow cytometry. The amount of lipofuscin in PG was assessed by the intensity of autofluorescence in organ frozen sections in 560-600 nm wavelength range. It was found that under the influence of MT, the number of intraorgan correlations in the immune system increased, regardless of the light regime. In animals on CL treated with MT, the number of interorgan connections was reduced, while negative correlations appeared between PG lipofuscin content and cellular composition of the spleen. The synchronizing and adaptogenic effects of MT were most pronounced under conditions of CL.

The study of the immune system of mice of CBA (n=7) and C57BL/6 (n=7) strains was performed during a 12-hour photoperiod (12:12) and round-the-clock illumination (RI) in December and March. The numbers of CD3+, CD4+8-, CD4-8-, CD4+8+, CD19+, CD3+hi lymphocyte subpopulations in the thymus and spleen, as well as the proliferative activity of thymocytes and splenocytes were measured by flow cytometry. It was found that the most pronounced. interstrain differences in the cellular composition of the thymus were observed in March, while those in the cellular composition of the spleen were detected in December. Overall, in C57BL/6 mice as compared with CBA mice, the number and proliferative activity of the cells of lymphoid organs was increased. In December, the influence of RI on the cellular composition of the thymus was more pronounced in C57BL/6 mice, while the effect on cellular composition of the spleen was greater in CBA nice. In March, after exposure to RI, a number of CD3+, CD19+ and CD4+8-splenocytes was increased in CBA mice, while it was decreased in C57BL/6 mice. In C57BL/6 mice, after exposure to RI, the proliferative activity of thymocytes decreased in winter and increased in spring . Interstrain and seasonal differences in the cellular composition of lymphoid organs in CBA and C57BL/6 mice, with a photoperiod of 12:12 and RI should be taken into account in experimental work with aninials of different genotypes in different seasons of the year.

It has been shown that psychoneurological disorders are accompanied by different disturbances of immunity. Paper aimed to study the effects of repeated experience of aggression in daily agonistic interactions leading to the development of behavioral psychopathology on the parameters of cellular immunity in the thymus and spleen. There were no found the changes in the weight indexes, the number of cells in the thymus, spleen and blood in aggressive mice. In the spleen of aggressive mice percent of B-lymphocytes--CD19+ and CD16/32+, as well as T-lymphocytes CD4+8-, CD4-8+, and CD4+25(hi) decreased and percent of CD4-25+ increased in comparison with the controls. In the thymus percent of CD4-25+ cells are decreased without changes of other types of lymphocytes. Flow cytometric analysis revealed decreased percentage of apoptotic (A(0)) and resting (G0/G1) cells and increased percentage of proliferating cells in phase S+G2/M in the spleen of aggressive male mice in comparison with the control. The percentage of apoptotic thymocytes is increased and the percentage of thymocytes in S+G2/M phase is decreased under the repeated experience of aggression. Data suggest the possible development of an autoimmune procceses in male mice under the influence of repeated experience of aggression.

Unlabelled: Chronic social defeat stress leads to the development of mixed anxiety-depression state, which accompanied by immune deficiency in male mice. Paper aimed to study effects of ronkoleukin on the parameters of cellular immunity in the thymus and spleen and psychoemotional state in these animals., Methods: Mixed anxiety/depression state was produced by chronic social defeat stress during 20 days in male mice. Roncoleukin (5000 ME/kg, i/p) and saline were chronically injected to depressive mice during 2 weeks without agonistic interactions. After this period subpopulations of lymphocytes in the thymus and spleen were studied in male mice. The animals were also studied in behavioral tests estimating the levels of communicativeness, anxiety and depressiveness., Results: Roncoleukin decreases the number of lymphocytes in the thymus and spleen, and increased the number of lymphocytes in blood and thymus index. Medication increased per cent of CD4+8+ lymphocytes in the thymus and per cent of CD8+ and CD3+25- lymphocytes in the spleen. Roncoleukin induced anxiogenic, stimulative and antidepressive effects., Conclusion: Roncoleukin has small efficacy for treatment of immune suppression induced by chronic social defeat stress and has anxiogenic, stimulating and weak antidepressive effects.

Aim: To study the influence of psychoemotional status on the development of experimental lung metastases of strain-specific murine Lewis lung carcinoma in C57BL/6J mice and hepatocarcinoma-29 in CBA/Lac male mice., Materials and Methods: Sensory contact model was used for generating animals with repeated experience of social victories or defeat in daily agonistic interactions. Tumor cells were injected into the tail vein after 20 days of aggressive confrontations and the number of metastases in the lung was calculated 16 days later., Results: The experimental metastasis is shown to develop differently in mice with opposing social experience: the winners of both strains had significantly less metastases in the lung than the losers., Conclusion: The results obtained indicate that psychoemotional status affects Lewis lung carcinoma and hepatocarcinoma-29 metastasis in male mice.

Subpopulation spectrum of T lymphocytes (CD3+, CD4+, CD8+, CD25+, and CD3(+)25+) in thymus, spleen and inguinal lymphatic nodes have been studied in male mice after 20 days of psychoemotional stress produced by social defeats in daily agonistic confrontations. A reduction of total number of cells, of absolute numbers of all researched subpopulations of lymphocytes and % CD3+ cells in thymus of submissive mice was shown in comparison with intact animals. Reduction of total number of splenocytes and absolute numbers of CD4+ and CD8+ lymphocytes has been observed in a spleen of submissive mice. Besides, % CD3+, CD25+, CD4+ and CD25+ cells were increased in these animals in comparison with intact mice. The absolute number of cells with CD8 phenotype was increased in inguinal lymphatic nodes. The data obtained suggest that the chronic psychoemotional stress is accompanied by serious changes of the cellular link of immunity. The effect of chronic emotional social stress on mutual interaction of the central and peripheral links of immunity has been discussed.

Chronic psychoemotional stress of social defeats produces development of experimental anxious depression in male mice similar to this disorder in humans. 5-HT and 5-HIAA levels, TPH and MAO A activities, 5-HT1A-receptors in different brain areas were investigated at different stages of development of experimental disorder. It has been shown that initial stage (3 days of social stress) is accompanied by increase of 5-HT level in some brain areas. Decreased 5-HIAA levels in the hippocampus, amygdala and nucleus accumbens were discovered at the stage of forming depression (10 days of social stress). Pharmacological desensitisation and decreased number of 5-HT1A-receptors were shown in frontal cortex and amygdala. At the stage of pronounced depression (20 days of stress), there were no differences in 5-HT and 5-HIAA levels in all brain areas (excluding hypothalamus) of depressive animals. However increased number of 5-HT1A-receptors and decreased affinity in amygdala and decreased TPH and MAOA activities in hippocampus were found in depressive mice. Hypofunction of serotonergic system is suggested at the stage of pronounced depression state in animals. Similar processes had place in brain dopaminergic systems. It is concluded that dynamic changes of brain monoaminergic activities accompany the development of anxious depression in animals. Various parameters of monoaminergic systems are differently changed depending on brain area, mediator system and stage of disorder.

The aim of this work was to examine the behavioral effects of a novel lithium-based enterosorbent, Noolit (665 mg/kg), on male mice with mixed depression/anxiety-like state evoked by exposure to repeated social defeats in daily agonistic confrontations. The lithium component allows Noolit to be used as a psychotropic drug. Two experiments are described, in which the therapeutic and preventative effects of chronic Noolit treatment were examined. Response to Noolit was assessed in the plus maze, open field, partition test, and Porsolt's test. In both experiments, Noolit produced obvious anxiolytic and antidepressant effects. Treatment with Noolit fully restored some behavioral parameters in the plus maze and open field in depressed mice and prevented depression that would otherwise have developed. It has been suggested that enterosorbent Noolit can be a potent drug for the treatment of mixed anxiety/depression pathologies and for prevention of mood disorders.

The binding of [3H]8-hydroxy-N, N-dipropyl-2-aminotetralin ([3H] 8-OH-DPAT) has been studied in various brain regions of male mice with the experience of repeated victory (winners) or defeat (losers) gained over 10 (T10) and 20 (T20) days of daily agonistic confrontations. The Bmax of [3H]8-OH-DPAT binding sites was found to be decreased in the hippocampus of the T20 winners when compared to the T10 winners or the control mice, and increased in the amygdala of the T20 losers. The Kd was increased in the amygdala of the T20 losers when compared to the animals of any other experimental group. No change in [3H]8-OH-DPAT binding was observed in the midbrain or the hypothalamus of the winners or losers. It was concluded that the 20-day agonistic confrontations, arranged in order to categorize the mice as winners or losers, had opposing effects on [3H]8-OH-DPAT binding.

The enterosorbent noolit (representing a mineral matrix base with immobilized lithium and silicon compounds) significantly affects the psychoemotional state of mice with depression model induced by prolonged social emotional stress. Noolit administration produces a pronounced anxiolytic and antidepressant effect and reduced the level of behavioral deficiency in the test animals.