氡与亚砷酸钠对HBE细胞联合毒性作用研究

发布时间：2018-06-21 22:54

本文选题：氡 + 亚砷酸钠　；参考：《苏州大学》2013年硕士论文

【摘要】：目的探讨氡与亚砷酸钠染毒永生化人上皮细胞（HBE cells）联合作用类型及联合染毒对HBE细胞氧化损伤和DNA损伤的影响。方法（1）以HBE细胞为研究对象，将HBE细胞单独暴露于20000Bq/m3的氡染毒不同时间（0～75min）、不同剂量（0～50000μmol/L）的亚砷酸钠以及二者的联合染毒，采用活细胞计数试剂盒(CCK-8)检测细胞存活率。通过中效原理计算不同时间氡暴露和不同剂量亚砷酸钠染毒联合作用的联合指数（combinationindex，CI）。（2）以不同时间氡暴露（20000Bq/m3，10、20和40min）和不同浓度亚砷酸钠（1.5、3.0和6.0μmol/L）处理HBE细胞，测定氡、亚砷酸钠单独及联合染毒后， HBE细胞活性氧（reactive oxygen species， ROS）、丙二醛（malondialdehyde，MDA）含量和超氧化物歧化酶（superoxi dismutase，SOD）活力的变化；通过彗星实验检测氡、亚砷酸钠单独及联合染毒后HBE细胞的DNA损伤情况；（3）通过Western blot检测在氡、亚砷酸钠单独及联合染毒后24小时，DNA损伤蛋白γH2AX和DNA修复蛋白Rad51。结果（1）氡与亚砷酸钠单独或联合染毒对HBE细胞均有抑制作用；不同时间氡暴露和不同剂量亚砷酸钠联合染毒的联合指数CI1，，表明二者呈现协同作用。（2）在单独染毒的条件下，与空白对照组相比，氡单独染毒组和亚砷酸钠单独染毒组ROS和MDA含量均随暴露时间延长和浓度增大显著升高，SOD活力均显著下降（P0.05）；且分别随着氡暴露时间的延长和亚砷酸钠染毒剂量的增加，HBE细胞内MDA和ROS含量均呈现明显的上升趋势，而SOD活力明显下降。当联合染毒时，分别在氡暴露时间恒定以及亚砷酸钠剂量恒定的条件下，各联合染毒组HBE细胞ROS和MDA的含量均明显增加（P0.05）；SOD活力均显著下降（P0.05）。（3）彗星实验：与对照组相比，氡单独染毒组及亚砷酸钠单独染毒组，各组HBE细胞的尾部DNA百分含量、尾距和Olive尾距均显著增加（P0.05）；当联合染毒时，在氡暴露时间恒定以及亚砷酸钠剂量恒定的条件下，联合染毒组HBE细胞尾部DNA百分含量、尾距和Olive尾距均显著增加，且均高于对照组、氡单独染毒组及亚砷酸钠单独染毒组（P0.05）。（4）Western blot实验：与对照组比较，各染毒组的DNA损伤蛋白γH2AX表达量明显增加，而DNA修复蛋白Rad51表达量下降；与氡单独染毒组和亚砷酸钠单独染毒组相比，联合染毒组γH2AX表达量明显增加，Rad51表达量明显降低，且随着染亚砷酸钠剂量的增加，γH2AX表达量呈现上升趋势，Rad51表达量呈现下降趋势。结论 1、氡与亚砷酸钠联合染毒HBE细胞具有协同抑制细胞生长的作用。 2、氡与亚砷酸钠联合染毒HBE细胞，通过造成严重的氧化损伤和DNA损伤，抑制其生长。
[Abstract]:Objective to investigate the combined effects of radon and sodium arsenite exposure on immortalized human epithelial cells (HBE cells) and the effects of combined exposure on oxidative damage and DNA damage of HBE cells. Methods (1) HBE cells were exposed to 20 000 Bq / m 3 radon for different time (0~75min), different doses (0 ~ 50000 渭 mol / L) of sodium arsenite and combined exposure. The survival rate of HBE cells was measured by living cell count kit (CCK-8). The combined index CI). (2 of radon exposure at different time and different doses of sodium arsenite was calculated by medium effect principle. HBE cells were treated with different time radon exposure (20000Bq / m31020 and 40min) and different concentrations of sodium arsenite (1.53.0 渭 mol / L and 6.0 渭 mol / L). The contents of reactive oxygen species (Ros), malondialdehyde (malondialdehyde) and the activity of superoxi dismutase (superoxi dismutase) in HBE cells were detected by comet assay. (3) the DNA damage protein 纬 H 2AX and DNA repair protein Rad51 were detected 24 hours after radon and sodium arsenite exposure alone or in combination by Western blot. Results (1) Radon and sodium arsenite alone or in combination had inhibitory effects on HBE cells, the combined index CI1 of radon exposure at different time and sodium arsenite at different doses showed synergistic effect. (2) under the condition of single exposure to radon and sodium arsenite, Compared with the blank control group, Ros and MDA contents in radon alone and sodium arsenite groups decreased significantly with the prolongation of exposure time and the increase of concentration (P0.05). With the prolongation of radon exposure time and the increase of sodium arsenite exposure, the contents of MDA and Ros in HBE cells showed an obvious upward trend, while the activity of SOD decreased significantly. Under the condition of constant radon exposure time and constant dose of sodium arsenite, the contents of Ros and MDA in HBE cells of each combined exposure group were significantly increased (P0.05), and the activity of). (was significantly decreased (P0.05) Comet assay: compared with the control group, the Ros and MDA contents of HBE cells in each combined exposure group were significantly higher than those in the control group. The tail DNA content, tail distance and Olive tail distance of HBE cells in each group were significantly increased in radon alone and sodium arsenite groups (P0.05), and under the condition of constant radon exposure time and constant dose of sodium arsenite, The percentage of tail DNA, tail distance and Olive tail distance of HBE cells in combined exposure group were significantly higher than those in control group. Western blot test of radon alone and sodium arsenite exposure group (P0.05). (4): compared with control group. The expression of DNA damage protein 纬 H 2AX increased significantly, but the expression of DNA repair protein Rad51 decreased, compared with radon alone and sodium arsenite alone, the expression of 纬 H 2AX in combined exposure group was significantly higher than that in sodium arsenite group, and the expression of 纬 H 2AX in combined exposure group was significantly lower than that in radon alone group and sodium arsenite group. With the increase of sodium arsenite, the expression of 纬 H 2AX increased and Rad51 decreased. Conclusion 1. HBE cells exposed to radon and sodium arsenite have synergistic effects on cell growth. 2. HBE cells exposed to radon and sodium arsenite combined with sodium arsenite can inhibit the growth of HBE cells by causing severe oxidative damage and DNA damage.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R114

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