| Objective:Immunity system is highly sensitive to ionising radiation and one of effector organs of electromagnetic radiation. In order to widen and deepen our knowledge ofγ-ray irradiation combined with microwave and provide experimental and theoretical data for prevention,diagnosis and therapy of its irradiation ,the injurious effects and mechanisms ofγ-ray irradiation combined with microwave on immunonity system were systematically investigated and the effects of some prevention measures determined.Materials and Methods:KM mice and Raji cells were divided into the following four groups: normal control, microwave(S frequency range,50mW/cm2),γ-ray irradiation (5.5Gy),γ-microwave combined radiation (5Gy+50mW/cm2).Using various methods such as biochemical tests, light microscopy ,inverted phase contrast microscope ,electron microscopy, flow cytometry , Annexin-V and PI double labelling,Fluo-3 fluorescent probe labelling ,laser scanning confocal microscope ,the injurious effects of 60COγ-ray irradiation combined with high power microwave on immunity system and their mechanisms were investigated in vitro and vivo at different levels of histology, cell and molecule. Moreover, To explore the injury effects and related mechanisms induced by microwave andγray on spleen and Raji cell.Results :The first part about alive mice: The main histopathological changes are apoptosis-necrosis, death, regeneration and recovery phase. Apoptosis and necrosis happen at 6 hours after radiation,regeneration and recovery occur at 7-28d after radiation.Injurious effects of spleen cell can be found in bothγ-ray irradiation group , S-HPM group andγ-microwave combined radiation group.The decrease in area ,perimeter,area density and circumference density have been observed in spleen bodies,and, the amount of lymphocyte cell reduce , perimeter,area density and circumference density of cell also lessen. The pathological change was more obvious and the recovery was slower inγ-microwave combined radiation group.The injury of S-HPM group show the increase of macrophage.The second part about cultured cell:①Cell morphology Normal Raji cell was small and round, bright and strong refraction. After radiation, the cell shape became irregular, cell body shrinked, and bad refraction. All the changes were most serious in groupγ+S-HPM, groupγwas the second, and group S-HPM was the most slight.②Cell cycle S phase: Cell number of each radiation group was more than control. Groupγandγ+S-HPM were significantly more than control (p<0.01), especially for groupγ+S-HPM. G0/G1 phase: Cell number of each radiation group was significantly less than control(p<0.01). There were no significant difference between radiation groups. G2/M phase: Cell number of group S-HPM was significantly more than control (p<0.05). Groupγandγ+S-HPM were less than S-HPM(p<0.01), numbers ofγ+S-HPM group go to G2/M phase were more thanγgroup, but no significant difference was seen between them(P>0.05)③Apoptosis and necrosis The rate of apoptosis and necrosis in radiation group were all higher than that of group control. There were significances between control and groupγorγ+S-HPM(p<0.05 or p<0.01), especially for groupγ+S-HPM (p<0.01). For radiation groups, groupγ+S-HPM was significantly more than group S-HPM andγ(p<0.01).④Ca2+ concentration The Ca2+ fluorescent intensity of cells in radiation groups were significantly stronger than that of group control (p<0.05 or p<0.01). There were significant difference between any two radiation groups(p<0.01). The Ca2+ fluorescent intensity was S-HPM <γ<γ+S-HPM by turns. Conclusion :Microwave andγray both affect the growth, proliferation, apoptosis and necrosis of lymphocyte. The injury effect ofγ+S-HPM compound radiation was more serious than any single radiation on lymphocyte. The major characteristics of injury showed asγray effect, and microwave aggravated the injury effect ofγray to some degree. Intracellular calcium overloading may be one of the injury mechanisms. |
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