An important factor in the development of inflammation is the presence in the body of lipopolysaccharide (LPS) – a component of the wall of gram-negative bacteria. LPS refers to pathogen-associated molecular patterns, which are a powerful activator of the immune system. The development of LPS-induced pathological processes can lead to reproductive dysfunction in women. Therefore, the search for and detection of new pharmacological strategies for the prevention and treatment of endotoxin-induced pathological disorders is extremely important.
The aim of the present study was to investigate the effect of ammonium glycyrrhizinate (GA) on changes in oogenesis, as well as the development of genotoxic stress, viability and death of lymph node cells under LPS-induced endotoxemia in female mice.
For determination of oocyte meiotic maturation the number of oocytes with germinal vesicle breakdown (metaphase I) was counted after 4 hours, while the number of oocytes forming the first polar body (metaphase II) was estimated by light microscopy after 20 hours of culture. The percentage of viable, apoptotic and necrotic lymph node cells was determined by their vital staining with fluorescent dyes Hoechst 33342 and propidium iodide. The degree of DNA damage was estimated by the method of alkaline single-cell gel electrophoresis (also known as Сomet assay).
The administration of GA caused an improvement of oocyte meiotic maturation, impaired by LPS treatment: the number of oocytes at metaphase I and metaphase II increased significantly compared to that of endotoxemic mice. The intraperitoneal application of LPS in mice increased significantly an amount of lymph nood cells with high degree of DNA damage (cometh 3 and type 4). The use of GA contributed to the reducing of LPS-induced DNA damage and weakening of genotoxic stress in the cells. Our data also demonstrated an improvement in the viability of lymphocytes, when GA was applied, that was occurred due to the weakening of their apoptotic and necrotic death.
Thus, our study showed that the introduction of GA had a pronounced anti-inflammatory and cytoprotective effect on lymph node cells which was manifested in the reduction of genotoxic stress and decrease in necrotic and apoptotic death of lymphocytes, as well as GA treatment exerted the stimulating effect on oocyte meiotic maturation under the conditions of experimental endotoxemia.
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