ABSTRACT
The work is devoted to the analysis of age-related changes in human spermatozoa and their functional properties in men aged 26-47 years. The work used ejaculate obtained from 54 men as part of the in vitro fertilization procedure. The patients were divided into three groups (26-29, 30-34 and over 35 years old). In the course of the work, the parameters of the spermogram were collected, as well as the results of assessing the viability of spermatozoa, obtained using the method of flow cytometry, and also a retrospective analysis of the effectiveness of in vitro fertilization was carried out. It was found that the average values of spermogram parameters in groups were within the physiological norm, however, about 59% of patients had individual deviations in terms of 1-3 indicators. Cytometric analysis revealed a rapid increase with age in functional disorders of spermatozoa, affecting the recognition and penetration apparatus (acrosome), the energy apparatus of the cell (its mitochondrion) and the density of chromatin in its nucleus. The result is a decrease in the probability of fertilization from 88% at 26-29 years old to 61% after 35 years, even with in vitro fertilization. The significance of the results obtained for the analysis of age-related changes in the male reproductive system and the practice of treating male infertility is substantiated.
Subject(s)
Fertilization , Infertility, Male , Acrosome , Humans , Male , Retrospective Studies , SpermatozoaSubject(s)
Apoptosis , T-Lymphocytes/cytology , Thymus Gland/immunology , Animals , Cell Count , Cell Proliferation , Flow Cytometry , Male , MiceABSTRACT
The ratio of early apoptosis and late apoptosis (necrosis) in the cultured human umbilical vein endothelial cells was estimated after exposure to hydrogen peroxide (H2O2) in vitro trying to keep them close to the physiological conditions (high cell density, high serum content, H2O2 concentration not over 500 µM). Cell viability was assessed using flow cytometry and simultaneous staining with fluorescent dyes PO-PRO-1 to detect early apoptotic cells, and DRAQ7 to detect late apoptotic and necrotic cells. The data obtained suggest that the primary mechanism of cytotoxic response is apoptosis. The critical concentration of H2O2 causing the death of the cell population in a dense monolayer is 250 µM. Lower concentrations of H2O2 (up to 200 µM) cause death of individual cells; however, viability of endothelial cell population is retained, and response to calcium activating agonists does not change compared with control cells.
