Bulletin of Experimental Biology and Medicine最新文献

Changes in the lipid and carbohydrate metabolism, adipokines, and growth factors during the development of metabolic disorders were studied in three mouse models: C57BL/6 (alimentary obesity), db/db (leptin-resistant obesity), and NOD (diabetes mellitus) lines. In the group of alimentary obesity, moderate fatty infiltration of the liver and hypertrophy of the adipose tissue, hyperglycemia, and increased concentrations of adiponectin, transforming growth factor β1 (TGF-β1), leptin, and cholesterol were detected. In the group of leptin-resistant obesity, multiple pathological changes in tissues, severe hyperglycemia and hyperleptinemia, hyperinsulinemia, and reduced concentrations of triglycerides, adiponectin, myostatin, and TGF-β1 were detected. In NOD mice, reduced number of insulin-positive β cells, hyperinsulinemia, and a decrease in adiponectin, TGF-β1, leptin, and myostatin concentrations were detected.

We studied the effect of acteoside on a model of human corneal epithelial cells (HCEC) injury induced by H₂O₂. HCEC were divided into 4 groups and cultured for 24 h in normal medium (intact and control groups, respectively), or in a medium containing DMSO or 160 μM acteoside (DMSO and acteoside groups, respectively). Then, H₂O₂ solution was added to HCEC for 4 h, except for intact cells. The cell viability was assessed by the CCK8 method to determine the working concentrations of acteoside and H₂O₂ for further experiments. Quantitative PCR and Western blotting were used to determine the effect of acteoside and H₂O₂ on the expression of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), NADPH:quinone oxidoreductase-1 (NQO-1), and cyclooxygenase-2 (COX-2). The effects of these agents on cell proliferation were assessed by staining with 5-ethynyl-2'-deoxyuridine (EdU). It was found that acteoside protected HCEC against H₂O₂-induced damage by inhibiting the expression of Nrf2, HO-1, NQO-1, and COX-2.

The involvement of DNA synthesis in the mechanisms of long-term memory reconsolidation in edible snails trained for conditioned food aversion was investigated. Administration of nucleoside analogs, such as 5-bromo-2'-deoxyuridine or 3'-azido-3'-deoxythymidine, which inhibit DNA synthesis, 1 h before or 1-3 h, but not 5 h after reminder with the conditioned stimulus led to memory impairment. One day after the inhibitor application and memory reactivation, a weakly expressed memory impairment (amnesia) was observed, which progressed over the next few days to the complete disappearance of behavioral memory expression. In snails with formed aversive memory for two food conditioned stimulus, a specific memory impairment was observed only for the stimulus that was paired with the presentation of an inhibitor during the reminder, while the memory for the other non-reactivated conditioned stimulus remained intact. It is suggested that DNA synthesis in the brain plays a specific role in the genetic mechanisms of reconsolidation and maintenance of long-term conditioned food aversion memory in snails.

The changes in astrocytes and their role in the development of brain diseases can be identified by morphological analysis of tissue specimens, in particular, by immunohistochemical detection of glial fibrillary acidic protein (GFAP). The study presents analysis of GFAP expression in white matter astrocytes of deceased newborns depending on the duration of the postmortem period. Autopsy material of the brain tissue obtained from 48 deceased newborns was divided into 8 groups depending on the duration of the postmortem period. Immunohistochemical analysis was performed with antibodies to GFAP in tissue samples taken from the superior and inferior brain areas relative to the position of the body stored before autopsy. The area of GFAP+ staining per field of view was determined in the white matter using an image analysis system. Morphometric analysis revealed a decrease in the mean values of the area of GFAP+ staining, i.e. the number of fibrotic astrocytes and their processes decreased with increasing the duration of the postmortem period. The mean areas of GFAP+ staining in the superior and inferior areas of the white matter did not differ significantly between the groups. The identified changes reflect the development of nonspecific postmortem changes which should be considered when taking tissue samples for molecular biological studies and in differential forensic diagnosis with lifetime lesions and diseases.

In adult male mice with high (HR) and low (LR) resistance to hypoxia, on days 21 and 28 after subcutaneous injection of Lewis lung carcinoma cells, a morphological and morphometric study of the primary tumor nodes and metastases in the lungs was carried out. Peripheral blood parameters and subpopulation composition of blood cells, the expression of genes responsible for the development of inflammation (Nfkb, Il1b, Il6, Tnfa, Il10, and Tgfb) and the response to hypoxia (Hif1a) in the liver were also assessed. The tumors were detected in 84.6% HR and 91.7% LR mice. The mitotic index of tumor cells in the subcutaneous nodes of HR animals was statistically significantly higher. The metastases area on days 21 and 28 did not differ. In animals of both groups, an increase in the absolute number of leukocytes, monocytes, and granulocytes, a decrease in the hemoglobin content and the absolute number of erythrocytes in the peripheral blood were detected on day 28 of the experiment. Only in LR animals, an increase in the absolute number of CD11b⁺ monocytes was found on day 28 of the experiment in comparison with the control group. The expression of Hif1a, Nfkb, Tnfa, and Tgfb genes in the liver of LR animals was higher than in HR mice, which attested to more pronounced systemic inflammatory response. These data should be taken into account when developing new approaches for the treatment of neoplastic disorders.

This work presents the method of synthesis and physicochemical characterization of isothiourea and cinnamic acid original derivative α-cyano-4-hydroxycinnamate 1-cyclohexanoy-l-2-ethylisothiourea (T1114). In studies of the cytotoxic and antitumor activity of T1114, it has been found that the combination in one molecular structure of NOS-inhibitory fragment (1-cyclohexanoyl-2-ethylisothiourea) and a fragment inhibiting monocarboxylate lactate transporters (MCT) (α-cyano-4-hydroxycinnamic acid) does not modify the cytotoxic activity of bifunctional NOS/MCT-inhibitor T1114 in vitro. But in vivo inhibition of NOS and MCT is able to realize effects on the tumor microenvironment and hypoxic tumor cells. Such structural and functional modification has significantly extended the antitumor activity of the new NOS/MCT inhibitor. The bifunctional compound not only realized a more pronounced antitumor effect, but also prevented the development of hypoxic adaptation in solid Ehrlich carcinoma and acquired the ability to overcome the resistance of mouse cervical cancer (RShM-5). Therefore, the combination of NOS-inhibitory, anti-vasculogenic and hypoxia-oriented toxic effects can create new opportunities in antiangiogenic therapy of malignant neoplasms.

In an experiment on Wistar rats, a Heymann's active nephritis model was reproduced. After the chronic course of the disease was confirmed, we compared the effectiveness of single systemic and local transplantation of allogeneic cultured stromal cells of the mononuclear fraction of the bone marrow. Both methods of cell therapy reduced clinical manifestations of active Heymann's nephritis: proteinuria decreased and glomerular filtration rate increased 30 days after cell administration. At the histological level, signs of the inflammatory process manifested in an increase in the number of CD68⁺ cells. Thus, in the case of autoimmune glomerulonephritis, two transplantation methods provide a comparable level of correction of functional disorders in Heymann's active nephritis model.

The functioning of the ventricular myocardium in hypertrophic cardiomyopathy (HCM) under high hemodynamic load leads to depletion of its resources and is associated with the risk of developing a dilated stage. In patients with HCM, the cardiomyocytes of the interventricular septum are hypertrophied, the proportion of cardiomyocytes in which myofibrils constitute less than 50% of the sarcoplasm volume increases with increasing the cardiomyocyte length and correlates with echocardiographic signs of left ventricular obstruction. These cardiomyocytes are characterized by ultrastructural signs of synthetic activity. In the myocardium of patients with HCM, single cardiomyocytes with "critical" loss of myofibrils and nonspecific degenerative changes corresponding to the picture of irreversible changes in the cardiomyocyte ultrastructure were revealed. The presence of cardiomyocytes with this ultrastructural phenotype is an early marker of exhaustion of the compensatory capabilities of the myocardium in HCM.

We studied the association of polymorphisms in the aminopeptidase N gene (ANPEP) with the development of diabetic retinopathy and nephropathy in patients with type 2 diabetes mellitus (T2DM). DNA samples from T2DM patients (n=1425) were genotyped for 23 single nucleotide polymorphisms (SNPs) using the MassARRAY system. Associations of SNP rs13380049 of the ANPEP gene with a lower risk of diabetic retinopathy (OR=0.54, 95%CI 0.36-0.82, p=0.0032) and nephropathy (OR=0.66, 95%CI 0.44-0.99, p=0.048) were found in T2DM patients. In addition, SNP rs25653 was associated with retinopathy, and polymorphisms rs6496608, rs72756574, rs9920421, and rs4932143 were associated with diabetic nephropathy. The study demonstrated the contribution of ANPEP gene polymorphisms in the determination of microvascular complications in T2DM patients.