放射损伤细胞中高尔基体弥散现象及香兰素衍生物的防护作用

摘要

目的：通过观察辐射对高尔基体形态的影响，确定香兰素衍生物VND3207对受照细胞高尔基体的防护作用。方法采用免疫荧光技术检测放射损伤细胞中高尔基体弥散现象并统计弥散面积，检测细胞周期变化，分析细胞存活率，同时对高尔基体蛋白的表达水平进行检测。结果免疫荧光检测结果显示，照射后高尔基体的弥散面积增加，具有一定的剂量效应和时间效应；VND3207能减轻γ射线对高尔基体的损伤，其表现为与未加药组相比，在抑制不同剂量照射后可使高尔基体弥散面积增加；细胞周期测定结果显示，4 Gy γ线照射后G2／M期阻滞峰值约出现在12 h后；免疫印迹结果显示DNA损伤引起的G2／M期阻滞也在12 h后解除；而高尔基体弥散在细胞周期阻滞解除后12和24 h仍然存在；平板克隆结果显示，VND3207促进了受照细胞的存活。结论放射损伤能引起剂量依赖性高尔基体弥散效应，且这种弥散与DNA损伤诱导的周期阻滞无关，VND3207对放射损伤引起的高尔基体弥散有一定的抑制作用，可能与受照细胞受到保护有关。%Objective To determine the Golgi dispersal in radiation damaged cells and the protective effect of vanillin derivatives.Methods Immunofluorescence, cell cycle analysis of flow-cytometry,Western blot,and clone formation were used.Results Immunofluorescence observation showed that the Golgi dispersal caused by 2 Gy 60 Coγ-ray was significantly increased in a dose-dependent manner in the range of 4-10 Gy as was demonstrated by the fact that the Golgi area was significantly increased. When the irradiated cells were treated with the radioprotective agent VND3207, a vanillin derivative,the Golgi dispersal induced by radiation was significantly reduced.The radiation-induced Golgi dispersal was also displayed in a pattern of time-course after irradiation in the HeLa cells, and persisted at least to 36 h post-irradiation. Cell cycle test results indicated that the Golgi dispersal was not associated with the G2/M arrest triggered by radiation-induced DNA damage response.VND3207 could promote cell survival by plate colony formation assay.Conclusion The Golgi dispersal can be caused byγ-ray irradiation in a dose-and time-dependent manner, and VND3207 can provide a good protection against radiation injury associated with inhibited Golgi dispersal.