Doklady Biochemistry and Biophysics最新文献

To search for substances selectively acting on tumor cells, phenotypic screening in a coculture of tumor cells with non-tumor cells was used in the work. The compound STOCK7S-36520, selectively cytotoxic in the coculture of breast tumor cells MCF7' and non-tumor MCF10A cells, contains structural elements characteristic of kinase inhibitors. Analysis of the compound STOCK7S-36520 and its derivative STOCK7S-47016 showed that they are new multikinase inhibitors. The highest inhibition of 84% was shown by compound STOCK7S-47016 against GCK kinase. Of interest is the significant selectivity of action against some of the cell lines studied: the selectivity index of STOCK7S-36520 against the prostate tumor cell line PC3 is 29 times compared to the model line of non-tumor fibroblasts VA13.

The aim of the study was to investigate the effect of photobiomodulation therapy (PBMT) on the amount of p53 in neurons of the pyramidal layer of the hippocampus after modeling septoplasty in rats. Materials and methods. Septoplasty was modeled in 48 Wistar rats. PBMT was administered to 24 rats in the early postoperative period for 2 to 6 days. Histological sections of the hippocampus were studied to determine p53-positive neurons on days 2, 4, and 6 after surgery in rats of both groups. Results. Compared with the control group (n = 5 rats), in both experimental groups there was an increase in the number of p53-positive neurons in the hippocampus, however, in the group where PBMT was performed in the early postoperative period after modeling septoplasty, the number of such neurons was lower, and in some subfields of the hippocampus on day 6 it even corresponded to the control group. Conclusions. The use of PBMT in the early postoperative period after modeling septoplasty helps to reduce stress-induced expression of the p53 protein in the pyramidal layer of the hippocampus in rats.

In recent years, therapeutic approaches against diabetes-induced liver damage have attracted great interest. Studies indicate the anticarcinogenic, anti-inflammatory, antioxidant, and lipid-lowering potential of hesperidin (HESP), a flavonoid in citrus fruits. This study examined how HESP prevented streptozotocin (STZ)-induced diabetic liver damage. Four groups of seven rats each were created: Control, HESP (100 mg/kg/day), STZ (45 mg/kg), and STZ + HESP (45 mg/kg and 100 mg/kg/day, respectively). Serum AST, ALT, LDH, LDL, triglyceride, total cholesterol levels, and the TNF-α, IL-1β, and caspase-3 expression levels of liver tissue in the STZ group were higher than the other groups (p < 0.05). However, these values were significantly lower (p < 0.05) in the STZ + HESP group compared to the STZ group. Furthermore, administering HESP together with STZ reduced liver expression levels of caspase-3, TNF-α, and IL-1β, as well as blood levels of AST, ALT, LDH, LDL, triglyceride, and total cholesterol. HESP against diabetes-induced hepatic damage reduced proinflammatory cytokine levels, and returned the lipid profile, and apoptotic indicators to normal levels. These findings suggested that HESP therapy may be an important therapeutic role against diabetes-induced liver damage.

Cisplatin (CIS) is widely used in the treatment of laryngeal cancer, one of the most common and lethal cancers. However, it is not a satisfactory chemotherapeutic agent. Therefore, there is a need to identify new agents, such as gallic acid (GAL), that can exert a synergistic effect to elucidate the pathophysiological mechanisms of the chemotherapeutic effects of CIS and to increase the effectiveness of treatment by preventing drug resistance. For this purpose, we investigated the stimulatory role of GAL on CIS-induced human laryngeal cancer (Hep-2) cell death via TRPM2 channel activation. For the study, four groups were formed from human laryngeal cancer (Hep-2) cells as Control, GAL (1OO μM), CIS (25 μM), and GAL + CIS. In the analyses made, cell viability, glutathione (GSH) and glutathione peroxidase (GSH-Px) enzyme activity, lipid peroxidation (LPx) levels, inflammation markers I-1β, IL-6, and TNF-α, Total Oxidant/Antioxidant (TOS and TAS) status, reactive oxygen species (ROS), caspase (Cas-3-9) activity, Transient Receptor Potential Melastatin 2 (TRPM2), and Poly Adp Ribose Polymerase-1, (PARP-1) levels in the cells were determined. CIS treatment caused laryngeal cancer cell cytotoxic and increased Cas-3-9, ROS, IL-1β, TNF-α, IL-6, TOS, LPx, TRPM2, and PARP-1 levels while decreasing cell viability, GSH-Px, GSH, and TAS levels. The combination of GAL and CIS treatment made the treatment even more effective. In conclusion, the increase in ROS and cell death levels mediated by TRPM2 activation in CIS Hep-2 cells was further enhanced by GAL treatment. Thus, CIS chemotherapy in Hep-2 cells may be enhanced by the synergistic effect of the GAL combination, and drug resistance may be reduced.

Previously, we created a modular nanotransporter (MNT) containing a monobody to Keap1, an intracellular protein inhibitor of the Nrf2 transcription factor that controls cellular protection from oxidative stress and is capable of interacting with Keap1 in hepatocytes and protect these cells from the effects of hydrogen peroxide. Oxidative liver damage by acetaminophen was used as a model to study the antitoxic effect of this MNT. Intraperitoneal injection of acetaminophen to mice resulted in an increase in the level of alanine aminotransferase and aspartate aminotransferase in the blood, as well as in liver edema. A significant decrease in the level of these enzymes in the blood, along with a decrease in liver edema, was observed after preliminary intravenous administration of MNT 2 h before the acetaminophen injection. The results obtained can be used as a basis for developing drugs to treat diseases associated with oxidative stress.

To test new antiviral drugs aimed at degrading the nucleocapsid protein (N-protein) of the SARS-CoV-2 virus, it is desirable to have cells expressing the N-protein, for which it is necessary to find conditions for the maximum achievable efficiency of cell transfection with a plasmid encoding this protein. For transfection, polyplexes were used consisting of a plasmid encoding the N-protein fused with the mRuby3 fluorescent protein and polyethyleneimine (PEI)-polyethylene glycol (PEG)-TAT peptide block copolymers. The dependence of the transfection efficiency of human lung adenocarcinoma A549 cells on the PEG/PEI and N/P ratios (the ratio of nitrogen in PEI to phosphate in DNA) was studied. Significant positive correlations were shown between transfection efficiency determined by flow cytometry, the N/P ratio, and the proportion of polyplexes sized 40–54 nm. The data obtained can serve as a basis for creating an animal model of lung cells transiently expressing the N protein of the SARS-CoV-2 virus.

Previously, polypeptide constructs—modular nanotransporters (MNTs)—were created to deliver biologically active molecules into the nuclei of melanoma cells. In the present work, polyethylene glycol (PEG) molecules were attached to them at the N-terminal cysteine, both with the possibility of their subsequent cleavage at the hydrolysis site of tumor-specific proteases, and without this site (non-detachable PEG). All MNT variants labeled with the radioisotope ¹¹¹In were administered to mice with inoculated Cloudman S91 melanoma. The kinetics of radioactivity distribution in the mouse body was studied using single-photon emission computed tomography. Analysis of the obtained data using a compartmental mathematical model allowed us to establish that the attachment of PEG to MNT increased its lifetime in the blood and significantly increased its accumulation in the tumor. Addition of a PEG detachment site by tumor-specific protease led to a strong retention of this MNT in the tumor. The data obtained can serve as a basis for the creation of new effective antitumor drugs.

ackground. Gestational diabetes mellitus (GDM) is a significant pregnancy complication characterized by elevated blood glucose levels. This condition arises from insulin resistance and impaired glucose metabolism during the gestational period. In this experimental study, we investigated the protective effect of avicularin against streptozotocin (STZ)-induced GDM in rats and explored the underlying mechanism.

Material and methods. Female Swiss Wistar rats were utilized in this study. Gestational diabetes mellitus (GDM) was induced in the rats through the administration of 25 mg/kg streptozotocin (STZ). The rats received oral administration of avicularin at doses of 5, 10, and 15 mg/kg. Body, placental, and fetal weights were measured. Blood glucose levels (BGL) and plasma insulin were assessed at various time intervals. Levels of glycogen, glycated hemoglobin (HbA1c), hemoglobin (Hb), visfatin, intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), antioxidants, lipids, inflammatory cytokines, apoptotic markers, and inflammatory parameters were evaluated. Additionally, mRNA expression was analyzed in hepatic tissue.

Results. Avicularin significantly (p < 0.001) improved the body weight, fetal body weight and reduced the placental weight. Avicularin significantly (p < 0.001) suppressed the elevated blood glucose level and improved the plasma insulin level. Avicularin significantly (p < 0.001) altered the levels of HbA1c, Hb, visfatin, ICAM-1, VCAM-1, LPO, CAT, GSH, GPx, SOD, GST, TNF-α, IL-1β, IL-6, Il-8, IL-10, IL-18, COX-2, PGE2, iNOS, NF-κB, Bax, Bcl-2, Bcl-2:Bax ratio and caspase-3 parameters. Avicularin treatment significantly (p < 0.001) suppressed the mRNA expression of TRL4, MyD88, NF-κB, NLRP3, RAGE, VCAM-1, MCP-1, VEGF, Nox2, and p65.

Conclusion. The results of this study demonstrated the protective effect of avicularin against streptozotocin-induced gestational diabetes mellitus via modulation of the TLR4/MyD88/NF-κB and Bax/Bcl-2/Caspase pathways.

Immunoassay (IA) methods performed in the wells of a polystyrene microplate are the basis of diagnostic studies. In the “sandwich” IA, a fundamentally important initial stage is the immobilization of anchor antibodies in the well of the plate, designed for specific binding of a given antigen from a biological fluid. One of the very promising options for antigen-recognizing molecules are single-domain antibodies (nanoantibodies, Nbs). The use of Nbs as anchor antibodies is hampered by their low efficiency of functioning after passive adsorption in the well of the plate. The development of a new format and immobilization method in the case of Nbs are fundamentally important for overcoming this problem. This work describes the development of a new format of an anchor bispecific nanoantibody (anchor-Nb) to improve the efficiency of both passive adsorption of anchor-Nb and subsequent stages of immobilization and detection of the target antigen in the well of a polystyrene plate.

Extracellular vesicles secreted by keratinocytes are involved in intercellular interactions and contain various proteins, mRNA and miRNA, lipids, due to which they can activate cell migration and proliferation. The secreted human protein SLURP-2 regulates growth and differentiation of epithelial cells and affects proliferation and migration of keratinocytes. In this work, we showed that extracellular vesicles secreted by keratinocytes in the presence of SLURP-2 stimulate migration of HaCaT keratinocytes. It was also found that the expression of miRNA-96 and miRNA-183, suppressing cell migration and proliferation, is decreased in the vesicles secreted by keratinocytes in presence of SLURP-2. Thus, it is shown that the stimulation of keratinocyte migration in presence of SLURP-2 is associated, in particular, with a change in the repertoire of extracellular vesicles secreted by these cells.