六溴环十二烷(HBCD)对人肝癌细胞HepG2的细胞毒性及机制

The cytotoxicity and mechanism of Hexabromoeyelododecane(HBCD) on HepG2 cells

为研究六溴环十二烷(Hexabromoeyelododecane,HBCD)对人肝癌细胞Hep G2的毒性效应及机制,设3个不同剂量HBCD试验组(10、15、20μg·m L^(-1))和溶剂对照组、阴性对照组及阳性对照组,将Hep G2细胞处理24、48、72 h后,用MTT(3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide)法检测细胞毒性;24 h后采用胞质分裂阻断法测定微核率;另在24 h后用细胞内2,7-二氢二氯荧光素(2',7'-dichlorofluorescin diacetate,DCFH-DA)法测定细胞内活性氧(Reactive oxygen species,ROS)的浓度.实验显示,HBCD对Hep G2细胞有毒性效应,且存在一定的时间和剂量效应,并导致Hep G2细胞的微核率和活性氧的浓度增高.研究表明,HBCD对Hep G2细胞有明显的细胞毒性.

For understanding the mechanism and the cytotoxicity of HBCD（Hexabromoeyelododecane）on HepG2 cells, we used the MTT assay,a cytokinesis-block micronucleus assay to evaluate the cytotoxicity of HBCD（10,15,20μg·mL^-1） on HepG2 cells as well as the control groups, and measured the fluorescent intensity of reactive oxygen species（ROS） using DCFH-DA. The results showed that the higher concentrations of HBCD reduced the cell survival rates. There were significant differences in the frequency of cytokinesis-block micronucleus between the groups. A significant increased frequency of cytokinesis-block micronucleus was observed in 10,15,20μg·mL-1 dose group compared with the negative control group（P〈0.05 and P〈0.01）. ROS concentration increased after HepG2 was exposed to HBCD for 24 hours compared with the control group（P〈0.01）. The results suggested that HBCD had cytotoxicity to HepG2 cells in vitro.