THE IMPACT OF THE CHEMICAL AGENTS OF DIFFERENT PHARMACOLOGICAL GROUPS ON THE KLOTHO PROTEIN CONCENTRATION IN THE CARDIOMYOCYTE AND NEUROCYTE SUSPENSION IN 120 MINUTE HYPOXIA IN VITRO.

Currently, central nervous and cardiovascular system disorders of hypoxia genesis are widely treated with drugs that restore blood flow, as well as drugs that affect cell metabolism, namely, individual units of adaptive molecular-biochemical reactions. Of particular interest are pharma-correction methods of affecting synthesis and expression of Klotho proteins, such as vitamin D and Necrostatin-1, estrogens. The purpose of the study was to identify the ability of vitamin D and tamoxifen estrogen receptor modulator to affect Klotho protein synthesis (under hypoxia in vitro modeling in brain and heart cells). The study was performed on sexually mature white non-linear rats - males weighing 190-230 g. Hypoxia in vitro was modeled by insertion of 0.6 μM concentration of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridil (MPTP) into the suspension of respiratory tract tissue differentiator cardiomyocytes. By means of immunoenzyme analysis, concentration of Nitrotyrosin (Ntz) (ELISA Kit "Hycult biotechnology b.v.") and Klotho protein (Elabscience, USA) in cell suspensions was evaluated. Statistical processing of the results was carried out using the STATISTICA® for Windows 6.0 program (StatSoft Inc., No. AXXR712D833214FAN5). The reliability of the differences was carried out using the Student's t-criterion. The data analysis demonstrated that a 120-minute MPTP incubation of cardiomyocytes and neurocytes resulted in a significant deficiency of Klotho protein concentration compared to intact suspensions. Such a decrease, in our opinion, is related to the development of oxidative stress in cell suspension (increase of Ntz by 65% ​​and 69% in the cardiomyocyte and neurocyte suspensions, respectively), as well as hyperproduction of proinflammatory cytokines such as tumor necrosis factor (TNF) and interferon (INF). Vitamin D (10-7) addition to the incubation medium of cardiomyocytes and neurocytes resulted in the increase of Klotho protein content by 56% on average, with 36% and 42% reduction of Ntz concentration, respectively. The registered effects of vitamin D are explained with its direct stimulating of the expression and synthesis of Klotho protein and limiting FGF23 hyperproduction. The introduction of tamoxifen estrogen receptor modulator (10-7) into the cardio- and neurocyte incubation medium did not affect Klotho protein concentration (an increase in concentration of 34% and 28% respectively) in advanced cell suspensions as much as vitamin D, however, nitrotyrosine concentration decrease made a more expressed impact, on average by 52% and 60%. The effects of tamoxifen are implemented due to its impact on the HSP proteins system, which provides the structural and functional integrity of the Klotho protein.