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卷 65, 编号 5 (2023)
Articles
Functional Interactions of BAF and LEM Proteins in the Formation of Germ Cells
摘要
Recovery of the nuclear structure after cell division requires special interactions between the integral proteins of the inner nuclear membrane having a special LEM domain (LEMD), nuclear lamina proteins (lamins) and the conserved BAF protein that serves as a central link in these interactions, providing topological relationships between chromatin and nuclear envelope. The dynamic transformations of these protein ensembles in the mitotic cycle are characterized in detail at the molecular level, however, less attention is paid to the developing germ cells undergoing meiotic divisions, despite of their nuclei, especially in diplotene oocytes, differ significantly in structure from the somatic nucleus. This review summarizes the still relatively scarce experimental data proving the significance of functional interactions between BAF and LEMD proteins for gamete formation, from the selection of germline cells to the transformation of haploid spermatids into morphologically mature spermatozoa.
407-419
The Derivation and Comparative Characterization of Mesenchymal Stem Cell Lines, Isolated from Human Pulp of a Deciduous Tooth of Children of Different Sexes
摘要
Two new non-immortalized fibroblast-like cell lines isolated from the pulp of Deciduous Tooth of Children of Different Sexes, named MSC-DP-1 and MSC-DP-2, were derived and characterized. In order to confirm the status of mesenchymal stem cells, a comparative analysis of a some characteristics in these lines was carried out at early and late passages. In the process of long-term cultivation, significant interline differences were revealed in the nature of replicative senescence (RS) and in growth characteristics. The MSC-DP-1 line was characterized by a later entry into the active stage of RS and more active proliferation compared to the MSC-DP-2 line. Karyotypic analysis showed that both lines have a normal human diploid karyotype at early passages. At the late (18th passage), in the stage of active PS, the MSC-DP-2 line also has a normal karyotype. And the MSC-DP-1 line, which enters the active stage of RS much later (at passage 42), has an abnormal karyotype with a large number of clonal chromosomal rearrangements. Both lines showed a high proportion of cells carrying antigens characteristic of human MSCs: CD44, CD73, CD90, CD105, HLA-ABC, and a low frequency of cells with CD34, CD45, and HLA-DR antigens. The cells of the derived lines at an early passage have the ability to differentiate in adipogenic, osteogenic and chondrogenic directions. But the MSC-DP-2 line shows weaker differentiation in the adipogenic direction, than the MSC-DP-1 line. In the process of RS, a significant weakening of adipogenic differentiation takes place in the MSC-DP-1 line, and it disappears in the MSC-DP-2 line. The activity of other differentiations does not change during RS. In general, the derived results confirm the status of MSCs for the derived lines and indicate interline differences in the RS process. However, the derived differences between these lines, as well as the comparison with the previously derived line MSC-DP do not indicate their gender nature. Apparently, they are associated with the genetic characteristics of different donors.
420-436
Study of EA.hy926 Endothelial Cells by Atomic Force and Scanning Ion-Conductance Microscopy
摘要
A two-section analytical system was developed and tested to study the culture of EA.hy926 endothelial cells in real time with high resolution imaging. Scanning ion-conductance microscopy was shown as more relevant method because it didn’t cause mechanical damage of cell, and made possible scanning on the membranes, when endothelial cells were surrounded by nutrient medium. The method allowed not only to analyze changes in the cells morphology, but also to identify extracellular (microfilaments) and intracellular (nucleolus) structures. The rigidity mapping showed that the rigidity of the endotheliocyte membrane varied from 357 to 796 Pa. After 240 min from the beginning of the observation, the formation of endothelial cells apoptotic bodies has begun, and the rigidity of the cell gradually increased, while rigidity of the apoptotic bodies decreased.
437-446
The Effect of Synthetic Polycation Poly-2-Dimethylaminoethylmethacrylate on Biological Activity of Mammalian Resident and Nonresident Cells
摘要
Cationic polymers are the positively charged macromolecules that have in their structure N-containing functional groups such as primary, secondary and tertiary amine groups; quaternary ammonium groups and others. The effect of synthetic polycation poly-2-dimethylaminoethylmethacrylate (PDMAEM) on biological activity of animal fibroblasts (CHL V-79 RJK) and human red blood cells (RBCs) was studied. The influence of PDMAEM on cell adhesion using fibroblast culture was analyzed. Cultural plastic treated or untreated by polycation was used as substrate. The polycation adsorption on polystyrene surface did not change the adhesive capacity of fibroblasts. Pretreatment of fibroblasts with PDMAEM did not influence at low concentrations (0.1 and 1 μg/mL) the adhesive properties of cells plated on the untreated plastic surface. At high concentrations (10 and 100 μg/mL) PDMAEM inhibited the attachment of fibroblasts to this substrate. Relationship between the inhibition of cell adhesion under PDMAEM action and the toxic effect on fibroblast viability has been found. The PDMAEM treatment of human RBCs at high doses led to the damage of cells and release of hemoglobin to incubation medium. At low doses PDMAEM practically did not influence the hemolysis of RBCs. It was shown that PDMAEM induced the change of the shape and aggregation of RBCs. The toxic effect of PDMAEM on human RBCs coincided as a whole with such effect for animal fibroblasts. Possible cell targets upon the PDMAEM effect are discussed.
447-460
Cytokine Profile of Myocardial Cells in Coronary Heart Disease and Ischemic Cardiomyopathy
摘要
In the present work, we carried out a comparative analysis of myocardial cytokine profile in patients with coronary heart disease (CHD) and in patients with ischemic cardiomyopathy (ICMP) associated with CHD. The concentrations of 41 cytokines secreted by 24-hour myocardial tissue culture intraoperatively sampled from the right atrial auricle (RAA, control) and peri-infarct left ventricular zone (PZ-LV) were determined by flow fluorimetry using a multiplex test system. The aim was to study in vitro cytokine profile of myocardial cells to search for possible predictors of adverse outcomes of surgical treatment of patients with CHD and ICMP. Myocardial secretion of proinflammatory molecules GM-CSF and IFN-γ increased significantly (up to 78-80 pg/g, p<0.05) in patients with ICMP associated with CHD in contrast to zero values in CHD. At the same time, there was a three-fold decrease in the concentration of fractalkin 3 ligand (Flt-3L; FMS-like tyrosine kinase 3 ligand). A decrease in Flt-3L secretion was observed in the PZ-LV in comparison with the RAA. In addition, compared with RAA, concentrations of fibroblast growth factor-2 (FGF-2), platelet-derived growth factor-AB/BB (PDGFAB/BB), interleukins IL-15 and IL-4, and a regulated upon activation, normal T cell expressed and secreted (RANTES; CCL5) were strongly reduced in PZ-LV myocardial tissue culture. Differences in the course of CHD and ICMP are discussed, and possible predictors of surgical treatment risk in patients of the two groups are suggested using correlation and regression analyses. Proinflammatory cytokines (IL-5, IL-6) and chemokines (Flt-3L, IL-8), as well as angiogenesis factors (VEGF) and angiostasis (IP-10), are proposed to be considered as potential markers of adverse outcome of surgical treatment of cardiovascular disease.
461-474
Effect of Glycodelin on the Level of T-Regulatory Lymphocytes and Acute Phase Proteins in Wistar Rats after Introduction of Allogenous Bone Marrow Cells
摘要
Amniotic variant of glycodelin (GdA) has pronounced immunomodulatory properties, participating in the formation of immune tolerance during pregnancy. We investigated the effect of glycodelin on the level of T regulatory lymphocytes (Treg) and the level of acute phase proteins (α-2-macroglobulin (α-2M), orosomucoid, C-reactive protein (CRP)) upon administration of allogeneic red bone marrow (BM) cells to Wistar rats in a dynamic experiment in vivo. It was found that the introduction of GdA in animal whith allogeneic BM led to an increase in the proportion of peripheral Treg among CD4+ lymphocytes at the end of the experiment (on the 21st day) in comparison with the group that was injected with BM. It was shown that glycodelin reduced the level of CRP and α-2M, but increased the level of orosomucoid in the serum of experimental animals at the beginning of the experiment (day 3), however, by the end of the experiment (day 21), normalization of protein values was observed in all groups of experimental animals acute phase to the level of intact animals. Thus, glycodelin is able to realize an immunosuppressive effect on allogeneic cells through an increase in the level of Treg and orosomucoid, as well as a decrease in the concentration of CRP and α-2M.
475-482
Immunohistochemical Study of Matrix Metalloproteinases 2 and 9 in the Left Ventricle of Preterm Rats Heart in the Late Postnatal Development
摘要
Immunohistochemical detection of matrix metalloproteinases (MMP) 2 and 9 was carried out in the myocardium of the left ventricle of prematurely born (21 and 21.5 days of gestation) and full-term (22 days of gestation) rats at 42, 56 and 180 days of the postnatal period of ontogenesis. The intensity of immunopositive staining was assessed by a scoring system. Preterm birth leads to an increase in the intensity of the immunopositive reaction to MMP-2 and MMP-9 in the wall of the left ventricle of rats. An increase in the intensity of the MMP-2 positive reaction in the left ventricle of rats is observed the earlier, the greater the degree of prematurity. The intensity of the MMP-9 positive reaction in the left ventricle of male rats is greater with a more pronounced degree of prematurity. An increase in the intensity of the MMP-2 and MMP-9 positive reaction in the wall of the left ventricle of female rats due to preterm birth is determined exclusively in animals born on the 21st day of pregnancy, that is, with a greater degree of prematurity.
483-489
Effect of Perinatal Hypoxia (Asphixia) on the Distribution of the α1 GABAA-Receptor Subunit in the Neocortex of Newborn Rats
摘要
The aim of this work was to study the distribution of the α1 GABAA-receptor subunit in the neocortical layers of rats in the neonatal period after exposure to hypoxia. The effect of hypoxia on the brain of newborn rats was carried out on the 2nd neonatal day for 1 h at an oxygen content in the respiratory mixture of 7.8%. An immunohistochemical reaction was used to detect the α1 GABAA-receptor subunit. The quantitative protein content was estimated from the density of immunostaining of the reaction product in the cytoplasm and processes of neurons. The somatosensory area of the neocortex was studied on the 5th and 10th neonatal days (P5, P10). It has been established that in the neocortex there is a significant population of young neurons containing the α1 subunit, which is part of the GABAA-receptor in the early stages of the neonatal period. By the end of the neonatal period in control animals, the staining density of the product of reaction to the detection of GABAAα1 in the layers of the neocortex increases significantly. Exposure to perinatal hypoxia causes a reduction in the number of neurons containing the α1 GABAA-receptor subunit and a significant decrease in the density of immune staining in all layers of the neocortex.
490-498
The Intensity of Degradation and Synthesis of Derma Intercellular Matrix in Rats after Local Cold Injury in the Presence of Metabolic Syndrome
摘要
Studies of tissue regeneration processes in the post-cold period are currently in demand in connection with the development of the Arctic, which is characterized by extremely low temperatures. Metabolic or hormonal abnormalities based on insulin resistance and compensatory hyperinsulinemia, which can lead to the development of cardiovascular diseases, and, consequently, to microcirculation disorders, are aggravating factors that increase the consequences of cold injuries. In this work, we studied the processes of degradation and synthesis of the intercellular matrix of the dermis, as well as the concentration of sialic acids in the blood serum after a local cold injury of the third degree. It was revealed that the regenerative process after local cold damage began on day 3 and was characterized by an increase in the collagen content of the dermis and the thickness of collagen fibers. Regeneration in the case of metabolic syndrome was accompanied by a slowdown in tissue repair processes. The course of the recovery process was associated with a change in the content of sialic acids in the blood serum, while a distinctive feature of the dynamics of this indicator was an increase in the concentration of sialic acids in metabolic disorders on day 7 and a slow slight decrease throughout the experiment, reaching the control values by day 21.
499-506