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Introduction. Metabolic syndrome (MS) can cause impaired spermatogenesis and a decrease in sperm counts. However, the details of the effect of MS on developing spermatogenic cells remain unclear. Difficulties in solving this problem, the inconsistency of published clinical data, indicate the advisability of using experimental models to solve this urgent problem of andrology and reproductology.The study objective is to describe to investigate the specifics of the course of meiotic prophase I and the activity of the processes of phagocytosis and autophagy in Sertoli cells of rats with experimentally induced MS and in the course of therapeutic and prophylactic procedures during the development of experimental MS.Materials and methods. The animals were divided into three groups, each of which included four sexually mature male rats: 1st group – males fed a standard diet; 2nd group – males receiving a diet high in fat and fructose for 60 days; 3rd group – males with MS receiving sulphate mineral waters therapy, low-intensity ultrahigh frequency electromagnetic radiation therapy. Testicular cells were examined using light and transmission electron microscopy. For the first time in animals with MS, an immunocytochemical study of the peculiarities of chromosome synapsis in prophase I of meiosis was carried out on the basis of analysis of spread synaptonemal complexes of meiotic chromosomes and immunocytochemical analysis of Sertoli cells and spermatogenic cells in squashed preparations of seminiferous tubules. The parametric Student’s t-test and the nonparametric Mann–Whitney U-test were used for statistical data processing.Results. As a result of a histological study of the structure of the seminiferous tubules of animals of three groups, a statistically significant decrease in the indices of the spermatogenesis index in 2nd and 3rd groups compared to the control was revealed. Immunomorphologically, in the spread nuclei of primary spermatocytes of rats of the 2nd and 3rd groups, violations of the architectonics of nuclei, the formation of synaptonemal complexes fragments and circular synaptonemal complexes, numerous atypical inclusions were found. Signs of pachytene arrest were found in 40–50 % of spermatocyte nuclei. In the study of squashed cells preparations of the seminiferous tubules of rats of the 2nd and 3rd groups, signs of phagocytosed synaptonemal complexes were found in the cytoplasm of Sertoli cells, which were confirmed using antibodies to the SCP3 protein. Thus, evidence for the phagocytosis of degenerating primary spermatocytes by Sertoli cells has been obtained. In Sertoli cells, spermatocytes and spermatids, many autophagosomes are found, using LC3B protein marker. The presence of autophagosomes in Sertoli cells and spermatogenic cells in animals of these two groups was also confirmed by electron microscopy. In male rats of the 2nd group, significant disturbances in the structure of the pachytene nuclei were revealed. In the cytoplasm of Sertoli cells and spermatids of rats of the 2nd group, lipid droplets, numerous phagolysosomes containing cell detritus were revealed. Structural damage and phagocytosis of mitochondria were found in Sertoli cells and spermatocytes. Аutophagy in Sertoli cells were most distinctive in animals of the 3rd group.Conclusion. In male rats with experimental MS, significant disturbances in the structure of the nuclei of meiotic cells, a high content of primary spermatocytes with signs of pachytene arrest were revealed. The results obtained are in good agreement with the data of other authors, who revealed a decrease in the number of spermatozoa in the epididymis of rats and mice when modeling MS. It is assumed that the activation of autophagy is an important factor in supporting the viability of Sertoli cells and supporting the viability of germ cells in stressful situations, including MS. Apparently, autophagy is an adaptive mechanism that removes the remnants of apoptotic spermatogenic cells that are selected as a result of MS development.
Introduction. Metabolic syndrome (MS) can cause impaired spermatogenesis and a decrease in sperm counts. However, the details of the effect of MS on developing spermatogenic cells remain unclear. Difficulties in solving this problem, the inconsistency of published clinical data, indicate the advisability of using experimental models to solve this urgent problem of andrology and reproductology.The study objective is to describe to investigate the specifics of the course of meiotic prophase I and the activity of the processes of phagocytosis and autophagy in Sertoli cells of rats with experimentally induced MS and in the course of therapeutic and prophylactic procedures during the development of experimental MS.Materials and methods. The animals were divided into three groups, each of which included four sexually mature male rats: 1st group – males fed a standard diet; 2nd group – males receiving a diet high in fat and fructose for 60 days; 3rd group – males with MS receiving sulphate mineral waters therapy, low-intensity ultrahigh frequency electromagnetic radiation therapy. Testicular cells were examined using light and transmission electron microscopy. For the first time in animals with MS, an immunocytochemical study of the peculiarities of chromosome synapsis in prophase I of meiosis was carried out on the basis of analysis of spread synaptonemal complexes of meiotic chromosomes and immunocytochemical analysis of Sertoli cells and spermatogenic cells in squashed preparations of seminiferous tubules. The parametric Student’s t-test and the nonparametric Mann–Whitney U-test were used for statistical data processing.Results. As a result of a histological study of the structure of the seminiferous tubules of animals of three groups, a statistically significant decrease in the indices of the spermatogenesis index in 2nd and 3rd groups compared to the control was revealed. Immunomorphologically, in the spread nuclei of primary spermatocytes of rats of the 2nd and 3rd groups, violations of the architectonics of nuclei, the formation of synaptonemal complexes fragments and circular synaptonemal complexes, numerous atypical inclusions were found. Signs of pachytene arrest were found in 40–50 % of spermatocyte nuclei. In the study of squashed cells preparations of the seminiferous tubules of rats of the 2nd and 3rd groups, signs of phagocytosed synaptonemal complexes were found in the cytoplasm of Sertoli cells, which were confirmed using antibodies to the SCP3 protein. Thus, evidence for the phagocytosis of degenerating primary spermatocytes by Sertoli cells has been obtained. In Sertoli cells, spermatocytes and spermatids, many autophagosomes are found, using LC3B protein marker. The presence of autophagosomes in Sertoli cells and spermatogenic cells in animals of these two groups was also confirmed by electron microscopy. In male rats of the 2nd group, significant disturbances in the structure of the pachytene nuclei were revealed. In the cytoplasm of Sertoli cells and spermatids of rats of the 2nd group, lipid droplets, numerous phagolysosomes containing cell detritus were revealed. Structural damage and phagocytosis of mitochondria were found in Sertoli cells and spermatocytes. Аutophagy in Sertoli cells were most distinctive in animals of the 3rd group.Conclusion. In male rats with experimental MS, significant disturbances in the structure of the nuclei of meiotic cells, a high content of primary spermatocytes with signs of pachytene arrest were revealed. The results obtained are in good agreement with the data of other authors, who revealed a decrease in the number of spermatozoa in the epididymis of rats and mice when modeling MS. It is assumed that the activation of autophagy is an important factor in supporting the viability of Sertoli cells and supporting the viability of germ cells in stressful situations, including MS. Apparently, autophagy is an adaptive mechanism that removes the remnants of apoptotic spermatogenic cells that are selected as a result of MS development.
Introduction. According to Russian Association of Urology a number of infertile couples ranges from 8 to 17.2 % in various regions of Russia. One of the causes of which is the male factor. Raising the age of the father reduces the chance of conception, increases the risk of early termination of pregnancy and the risk of certain diseases of the child. In this regard the existing knowledge of age-related changes in ejaculate parameters needs to be de veloped. This is particularly true for men of infertile couples.The study objective – to investigate the age patterns of change in the spermological status of infertile men and identify the most age-related variations.Materials and methods. Research design – observation, retrospective, crossection. Analysis of the medical records of 517 Novosibirsk male residents from couples with infertility in marriage was made. The sampling is divided into 5 age groups: 1st group (n = 108) – men of ≤29.9 years old; 2nd group (n = 171) – 30.0–34.9 years old; 3rd group (n = 130) – 35.0–39.9 years old; 4th group (n = 68) – 40.0–44.9 years old; 5th group (n = 40) – ≥45.0 years old.Results. Median of male age in total sample – 34 years, frequency of persons over 40 years – 21 %. Overall sample shows a decline in reference values of proportion of morphologically normal forms – 3.0 (1.5–5.0) % and increase in sperm DNA fragmentation index (SDFI) – 16.0 (11.0–22.2) %. In groups from 1st to 5th there is an age-specific decrease in ejaculate volume (3.5 (2.5–4.8) ml; 3.3 (2.4–4.2) ml; 3.2 (2.4–4.5) ml; 3.0 (1.7–4.9) ml; 2.7 (1.9–3.5) ml, р1–5 <0.005) and sperm count (127.1 (58.8–264.0) million; 122.5 (62.4–214.0) million; 118.5 (52.8–217.5) million; 98.0 (58.5–199.5) million; 81.0 (36.5–137.9) million, р1–5, 2–5 <0.005). SDFI has an age-associated increase in the value of the indicator (14.4 (10.8–19.5) %; 15.0 (10.0–21.0) %; 17.2 (11.0–22.0) %; 18.5 (14.0–24.2) %; 19.2 (13.2–29.6) %, р1–4 <0.005).Conclusion. It was first time shown for the male population of Novosibirsk of Siberian Federal District that a reduction in the proportion of morphologically normal sperm below the reference values observed in the total sample and it did not depend on the age of the men examined. Only three of the examined indicators of sperm status studied infertile males had a significant age dependency: ejaculate volume decrease, reduction of total sperm, increased DNA fragmentation of sperm. The lack of a reliable influence of age on HBA-test indicators was first shown. However the trends identified require further study. The results show that age-associated increase in DNA fragmentation of sperm starts with young men under 30, and underscores the need to define SDFI in actual clinical practice. And negative processes of age influence on the quantitative, morphological and ultrastructural characteristics of sperm occur most frequently in men after 40 years.
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