Vol 65, No 1 (2023)

A review of current literature data on the significance of the Notch signaling pathway in the mechanisms of the development of diseases of the respiratory system – chronic obstructive pulmonary disease (COPD), bronchial asthma (BA) and lung cancer is presented. In studies of lung tissue samples of patients with COPD and lung tissues of mice, it was found that activation of the Notch signaling pathway promotes metaplasia and increases the functional activity of goblet cells, protects epithelial cells from apoptosis and oxidative stress. Suppression of the Notch−Jagged1/Jagged2 pathway is associated with the transdifferentiation of club-shaped cells into ciliated ones. In patients with AD, the Notch signaling pathway promotes differentiation of Th2 lymphocytes. In the ovalbumin-induced bronchial asthma model, the Notch cascade increases the imbalance of Th17/Treg lymphocyte populations, the production of IL-4, IL-5, IL-13, IL-17, the formation of allergen-specific IgE, eosinophilic infiltration and metaplasia of goblet-shaped epithelial cells of the respiratory tract. A decrease in the concentration of IgE, Th2-type cytokines (IL-4, IL‑5, IL-13), an increase in the number of Treg cells and the level of TGFß in bronchoalveolar lavage in mice with asthma, mediated by the introduction of dendritic cells expressing the ligands DLL1 and Jagged1, indicates the protective role of the Notch signaling pathway. On samples of tumor tissue and cell lines of non-small cell lung cancer, it was found that an increase in the expression of Notch-1 and Notch-3 mRNA is associated with increased proliferative activity, malignant cell transformation, a high risk of metastasis to lymph nodes and an unfavorable prognosis of the disease. In the samples of tumor tissue of small cell lung cancer, an increase in the expression of the Notch ligand DLK1 signaling inhibitor gene, the Ascl1 transcription factor gene and lysine-specific histone demethylase 1 (LSD1) was recorded. Suppression of LSD1 activity is accompanied by reactivation of signaling via Notch-1 receptor and subsequent inhibition of the transcription factor Ascl1, which induces the initial stages of tumor transformation.

Fracture healing is a complex process in which the periosteum and endosteum become the main sources of osteoblast progenitor cells. However, cellular mechanisms and signaling cascades underlying the early stages of osteoblast progenitors differentiation in adult bone are still not well understood. Therefore, we performed shotgun proteomics analysis of primary culture of isolated human osteoblasts from femur of adult donors in undifferentiated conditions and on the sixth day of osteogenic differentiation in vitro. This is an early timepoint in which we have observed no extracellular matrix mineralization yet. 1612 proteins identified with at least two unique peptides were included in proteomics analysis. Data are available via ProteomeXchange with identifier PXD033697. Despite the fact, that matrix mineralization starts only after induction of osteogenic differentiation, we revealed unexpectedly weak physiological shift associated with a decrease of cells proliferative activity and changes in proteins inVved in extracellular matrix secretion and organization. We demonstrated that osteoblasts were positive for markers of later osteogenic differentiation stages during standard cultivation: osteopontin, osteocalcin, BMP-2/4 and RUNX2. Therefore, further differentiation required for matrix mineralization needs minimal physiological changes.

The treatment of solid tumors with a cold atmospheric plasma jet (CAP) is an innovative approach, which began to be actively developed only in the last decade. As a consequence, the studies aimed at revealing the conditions of selectivity of such effects on tumor cells, including in 3D tumor models, are important. It is known that the main cytotoxic effects of CAP are caused by reactive oxygen and nitrogen species, which are formed in the plasma flow and the availability of which for the cells in the classical 2D and 3D cultivation models may be different. We used multicellular spheroids of MCF7-EGFR cells with hyperexpression of epidermal growth factor receptor (EGFR), the parental MCF7 breast adenocarcinoma cell line, and MCF10A non-transformed human breast cells. Irradiation of MCF7-EGFR spheroids led to destruction of multicellular 3D structures into individual cells with activation of death processes. It was shown that cells of CAP-irradiated spheroids underwent phagocytosis by activated macrophages. When comparing direct exposure to CAP and cultivation of MCF7-EGFR spheroids in CAP-irradiated medium (CAP-IM), a higher content of reactive oxygen and nitrogen species in spheroid cells was found when cultured in CAP-IM, which further leads to a greater cytotoxic effect than in direct irradiation. The cytotoxicity of CAP-IM has been shown to be valid longer when such medium is stored at 4 than at −20°С. Thus, it was shown that the treatment of spheroids with CAP-IM was more effective in death induction than direct CAP irradiation.

Decreased energy metabolism in the brain correlates with cognitive impairment in Alzheimer’s disease. Accumulating experimental data indicate that lactate transporters and monocarboxylate transporters (MCTs) are directly involved in cerebral energy metabolism. However, to date, changes in lactate levels and MCT content in Alzheimer’s disease remain unclear. The aim of the study was to study the content of lactate and of its transporters – MCT1 and MCT2 in cells of neuronal, astroglial and endothelial nature under acute toxic effects of beta-amyloid (Aβ1–42) in vitro and in vivo. Under conditions of acute toxic action of Aβ1–42 in vivo, a significant (P ≤ 0.05) decrease in the level of lactate in the hippocampal tissue and an increase (P ≤ 0.05) in the dialysate were found. At the same time, a low (P ≤ 0.05) levels of MCT1 and MCT2 was set. In vitro, significantly high (P ≤ 0.05) production of lactate by astrocytes was revealed, coupled with low (P ≤ 0.05) level of MCT2 on neurons. Thus, it was found that Aβ1–42 causes a decrease in the level of lactate in the hippocampal tissue and an increase in its level in dialysate in vivo, which correlates with the impaired level of MCT1 and MCT2. This indicates a violation of energy metabolism due to the acute toxic effect of Aβ1–42. At the same time, the revealed increase in the production of lactate by astrocytes in vitro may indicate the inclusion of a compensatory mechanism aimed at maintaining the astrocyte-neuronal interaction.

Analysis of the participation of short peptides GER and FGER containing common tripeptide fragment in the regulation of adhesive response of CHO-K1 cells was conducted. Both peptides stimulated cell adhesion both to untreated plastic and to gelatin-coated plastic, but did not change cell attachment to poly-L-lysine-coated plastic. Tripeptide GER had larger stimulation effect on cell adhesion to untreated plastic. Peptide FGER increased the rate of cell attachment to gelatin in a wider range of concentrations as compared to adhesion to untreated plastic. Variativity of cell spreading to different substrates under peptide action was demonstrated. On untreated plastic both investigated peptides practically in equal extent stimulated cell spreading. On gelatin peptide FGER kept the stimulation effect on cell spreading, but peptide GER partly inhibited cell spreading as compared to cell spreading on untreated plastic. It was established that insertion of additional N-terminal hydrophobic amino acid residue Phe to tripeptide fragment GER changes the regulatory activity of peptide at the cell adhesion model depending on the stage of cell connection with substrate and/or on substrate properties. The structural-functional activity of investigated short peptides on the instance of different structural components of adhesive structures is discussed.