X射线致HeLa细胞损伤过程中NADPH氧化酶的作用

杜斌斌; 刘箐; 张维胜; 王晓妍; 贾鹏飞; 王小虎; 李莎; 杨熊飞; 张红; 郭天康; #

doi:10.11804/NuclPhysRev.25.04.391

X射线致HeLa细胞损伤过程中NADPH氧化酶的作用

Role of NADPH Oxidase in HeLa Cell Lesion Induced by Xray Irradiation

摘要

摘要: 以HeLa细胞为实验材料，探讨了NADPH氧化酶在X射线诱导细胞损伤过程中的作用。结果显示， 12 Gy X射线辐照后细胞内活性氧(ROS)明显增加，在用NADPH氧化酶抑制剂处理后再辐照，则细胞内ROS降低到未辐照水平；同时辐照后NADPH 氧化酶细胞质亚基p47^phox 在细胞质积聚并和细胞膜亚基 gp91phox 结合； Western blotting检测结果显示， NADPH 氧化酶的关键亚基 gp91phox 的表达量明显增加。以上结果说明， NADPH氧化酶可以被X射线激活，由其介导产生的ROS在X射线诱导HeLa细胞损伤过程中扮演重要角色。 To investigate the role of NADPH oxidase in HeLa cell lesion induced by Xray irradiation, the change of cell survival was detected with MTT assay， reactive oxygen species (ROS) was measured by fluorospectrophotometer. Immunostaining and confocal laserscanning microscopy was employed to detect the colocalization of two subunit of NADPH oxidase, p47^phox and gp91^phox in the cell. Western blotting was used to detect the expression of gp91^phox before and after Xray irradiation. After Xray irradiation， intracellular level of ROS increased obviously. But the increase could be blocked by diphenyleneiodonium (DPI)， an inhibitor of NADPH oxidase. Meanwhile， cytosolic subunit p47^phox moved to membrane and colocalizated with gp91^phox after
irradiation. Moreover, the results also show that gp91^phox increased sharply after 12 Gy Xray irradiation. Therefore， NADPH oxidasemediated production of ROS plays an important role in HeLa cell lesion induced by Xray.

Abstract: To investigate the role of NADPH oxidase in HeLa cell lesion induced by Xray irradiation, the change of cell survival was detected with MTT assay， reactive oxygen species (ROS) was measured by fluorospectrophotometer. Immunostaining and confocal laserscanning microscopy was employed to detect the colocalization of two subunit of NADPH oxidase, p47^phox and gp91^phox in the cell. Western blotting was used to detect the expression of gp91^phox before and after Xray irradiation. After Xray irradiation， intracellular level of ROS increased obviously. But the increase could be blocked by diphenyleneiodonium (DPI)， an inhibitor of NADPH oxidase. Meanwhile， cytosolic subunit p47^phox moved to membrane and colocalizated with gp91^phox after
irradiation. Moreover, the results also show that gp91^phox increased sharply after 12 Gy Xray irradiation. Therefore， NADPH oxidasemediated production of ROS plays an important role in HeLa cell lesion induced by Xray.

HTML全文

参考文献(0)

施引文献

资源附件(0)