微波和γ线复合照射致Raji细胞损伤的形态和计量学研究

Morphometric study on the Raji cell damages induced by combination of gamma ray and microwave radiation

目的研究高功率微波和γ线复合照射引发Raji细胞凋亡和坏死发生规律及Ca2+浓度变化在其中的作用。方法建立微波和γ线复合照射Raji细胞模型;细胞分成4个实验组,单纯S-HPM照射组(S-HPM组)、单纯60Coγ射线照射组(γ组)、微波和γ线复合照射组(γ+S-HPM组)及实验对照组,采用Annexin-V和PI双标记流式细胞术检测细胞凋亡和坏死率的改变;利用Fluo-3荧光探针负载和激光扫描共聚焦显微镜(LSCM)定量检测细胞内游离Ca2+浓度的变化。结果1.细胞凋亡和坏死:γ线与微波复合照射后出现典型的凋亡和坏死改变,γ线组和复合组与对照组比较凋亡率和坏死率显著升高,同单纯γ线组相比,复合组凋亡和坏死率升高更显著。2.Ca2+浓度:各辐射组细胞内Ca2+荧光强度均明显高于对照组,各辐射组间均具有显著性差异,强度依次为γ+S-HPM>γ>S-HPM。结论单纯微波和γ线照射均可导致Raji细胞凋亡和坏死率增加,微波与γ线复合照射的损伤效应较单纯照射加重,其损伤特点主要表现为γ线效应,微波在一定程度上加重了γ线的损伤;胞内钙超载可能是其损伤机制之一。

Objective To investigate the apoptosis and necrosis in Raji ceils induced by combined microwave and γ ray irradiation and the association with intracellular Ca2. concentration changes. Method The cells of Raji cell line were used in this experiment and divided into 4 groups： S-HPM irradiation alone, γ ray irradiation alone, γ＋ S-HPM, and control groups. Annexin-V and PI double labeling were used to detect the apoptosis and necrosis in the Raji cells. Fluo-3 fluorescent probe labeling and laser scanning confocal microscopy were applied to examine the changes of intracellular free Ca^2＋ concentration. Results 1. Apoptosis and necrosis： γ ray combined with microwave irradiation induced apoptosis and necrosis in Raji cells. The amount of apoptotic and necrotic cells in both γ ray irradiation group and 7 ＋ S-HPM group were significantly more than those of control group. Moreover, the amount of apoptotic cells in the combined radiation group was significantly more than that in γ ray irradiation group. 2. Ca^2＋ concentration： The Ca^2＋ fluorescence intensities in cells of S-HPM, γ and γ ＋ S-HPM radiation groups were significantly stronger than that of control group, There was significant difference between any two radiation groups. The Ca^2＋ fluorescence intensity was in a order of S-HPM 〈 γ〈 γ ＋ S-HPM. Conclusion Both microwave and γ ray increase apoptosis and necrosis in Raji cells. The injury effect of γ＋ S-HPM combined irradiation is more serious than any single irradiation on Raji cells. The major injury dipicted under conditions of this experiment is caused by γ ray irradiation, and further enhanced by microwave irradiation. Intracellular calcium overloading may be one of the injury mechanisms.