АНАЛИЗ ПОЯВЛЕНИЯ МИКРОЯДЕР В ЭРИТРОЦИТАХ И АКТИВНОСТИ ПРОЛИФЕРАЦИИ КЛЕТОК КОСТНОГО МОЗГА ПОСЛЕ ПРОЛОНГИРОВАННОГО ОБЛУЧЕНИЯ МЫШЕЙ БЫСТРЫМИ НЕЙТРОНАМИ В НИЗКИХ ДОЗАХ

Abstract

Purpose: To analyze the level of cytogenetic damage and the activity of bone marrow cells proliferation in C57BL/6 mice after prolonged fast neutrons low dose irradiation at 10–500 mGy. 
 Material and methods: Male C57BL/6 mice at the age of 7–8 and 16 weeks were used in the experiments. Irradiation was carried out on an OR-M installation in the field of fast neutrons and gamma quanta using five Pu(α,n)Be radionuclide sources with a high fast neutron yield at a dose rate of 2.13 mGy/h. The frequency of polychromatophilic (PCE) and normochromic (NCE) erythrocytes with micronuclei (MN) and the ratio of PCE and NCE were analyzed using light microscopy after cytochemical staining of the bone marrow cells of control and irradiated mice. The proliferation activity of bone marrow cells was determined by the number of Ki-67+-cells. The parameters of the cell cycle and the level of apoptosis were studied after DNA staining with DAPI using flow cytometry. Statistical processing of the results was carried out according to the Student’s method using the computer program Origin.
 Results: It was found that prolonged irradiation of mice with fast neutrons at a low dose rate (2.13 mGy/h) at doses from 10 to 500 mGy after 24 h led to statistically significant increase in the frequency of PCE with MN at all studied doses. No dose dependence of this parameter was observed in the studied range. The increase in the frequency of PCE with MN at a dose of 500 mGy was prolonged and persisted for at least 72 h. A significant increase in the frequency of NCE with MN 24 h after irradiation was found only at a dose of 500 mGy, which persisted up to 48 h. At this dose, there was also a decrease in the number of nucleated cells in the bone marrow 24 – 72 h after exposure, a decrease in the number of Ki-67+-cells 24 h after irradiation of mice, a block of the cell cycle in the G2/M phase, and a decrease of cells in the G0/G1 phase, but after 48 h, there were no disturbances in the cell cycle. 
 Conclusion: It has been shown that after a single total prolonged irradiation of mice at low doses (10–500 mGy), when analyzing the frequency of PCE with MN, cytogenetic damage is recorded in the bone marrow, which indicates the genetic danger of exposure to even such low levels of fast neutron irradiation. A decrease in Ki67+ cells and cell cycle arrest at the G2/M phase were found only after irradiation of mice at a dose of 500 mGy and only 24 h after exposure, while the number of nucleated cells in the bone marrow at this dose was reduced, at least to 72 h.