Macrophage metabolic plasticity providing their polarization towards classically (M1) or alternatively (M2) activated cells is an important element of the initiation, development, and resolving or inflammation-linked pathologies. The prevalence of M1 or M2 types of macrophages during different stages of diseases supports increased inflammation and phagocytosis or tissue repair, respectively. An imbalance leading to a shift toward an M1- or M2-dominant state is associated with a chronic pathological process. This characterizes the regulation of macrophage phenotypes as a prospective strategy in the treatment of various diseases and makes it relevant to a deep understanding of the mechanisms defining cell polarization. According to the central role of calcium signaling in cell metabolism, changes in calcium homeostasis are closely linked to the regulation of polarization. The exact balance between calcium flows across plasma and intracellular membranes provided by a number of receptors and channels, as well as the differences in the calcium-buffering capability of endoplasmic reticulum and mitochondria, are able to influence macrophage polarization towards an M1 or M2 phenotype. This review focuses on the role of the calcium homeostasis system in macrophage functionality and calcium-induced changes in macrophage metabolism that forms the basis of target disease therapy.

BACKGROUND: Currently, there is no unified approach or effective method for treating premature ovarian insufficiency. The primary strategy is hormone replacement therapy aimed at mitigating estrogen deficiency and its associated complications. However, this therapy does not restore lost ovarian function or fertility. Thus, further research into the pathogenesis of premature ovarian insufficiency is crucial for developing alternative pathogenetically based therapies. Investigating the efficacy of various drugs in preclinical trials using cellular models holds significant promise. Experimental modeling of premature ovarian insufficiency, which closely replicates the origin and development mechanism of the human disease, can be effectively used to develop promising therapeutic approaches, in particular, for testing new drugs. AIM: The aim of this study was to develop a new method for experimental modeling of premature ovarian insufficiency using cyclophosphamide in Wistar rats, the significant advantages of which are high reproducibility, ease of implementation, and cost-effectiveness. MATERIALS AND METHODS: A culture of Wistar rat ovarian granulosa cells after five stages of subculturing was treated with the drug cyclophosphamide, ensuring a working concentration in the growth medium of 0.1 mg/ml, followed by incubation for six hours. RESULTS: A cellular model of premature ovarian insufficiency has been created, which is characterized by 100% modeling efficiency, high manufacturability and environmental safety for modeling the pathological condition. CONCLUSIONS: The model created will allow for testing the medicinal effectiveness of chemicals with a view to their further use in medicine.

Background: Premature ovarian insufficiency is a syndrome characterized by secondary hypergonadotropic ovarian insufficiency and a decrease in the ovarian function in women under 40 years of age, thereby leading to impaired reproductive function, metabolic changes, and a decrease in the quality of life. Increased production of reactive oxygen species inhibits the mitochondrial respiration chain and leads to mitochondrial dysfunction and oxidative stress, which are considered as the triggers of premature ovarian insufficiency. To date, there is no universal method of premature ovarian insufficiency prevention, and accepted treatment methods can only compensate for clinical symptoms, but not restore the lost ovarian reserve. Aim: The aim of this study was to develop a new method of premature ovarian insufficiency treatment based on an experimental ovarian granulosa cell model using omaveloxolone (RTA 408). Materials and methods: The cell model of premature ovarian insufficiency used in the study was realized according to the Patent RU 2 815 539 C1, 2023 (by M.I. Yarmolinskaya, K.A. Zakuraeva, A.Yu. Vinokurov, M.Yu. Pogonyalova). Ovarian granulosa cells isolated from three-month Wistar rats were subcultivated five times with subsequent seeding on coverslips. The cells on the coverslips were divided into three groups (three coverslip in each group). In Group 1 (comparison group — premature ovarian insufficiency model without treatment), cells were treated with cyclophosphamide (0.1 mg/ml) during 6 h. In Group 2 (main group — premature ovarian insufficiency model with experimental treatment) cells were pretreated with RTA 408 (2 µl/ml) during 1 h with subsequent addition of cyclophosphamide (0.1 mg/ml) and cultivation during 6 h. And in Group 3 (control group), cells were added no substances. Results: Compared to Groups 2 and 3, the use of omaveloxolone (RTA 408) led to an increase in reduced glutathione level and a decrease in reactive oxygen species production rate, which indicates the antioxidant and anti-inflammatory effects of the drug and may be considered as a perspective strategy of premature ovarian insufficiency treatment. The claimed method expands the number of premature ovarian insufficiency treatment strategies and avoids the use of hormonal medications and surgical procedures, thus reducing the risk of side effects and complications associated with their use.