Cui Yuan, Liu Wei, Xie Wenping, Yu Wenlian, Wang Cheng, Chen Huiming
Chinese Academy of Inspection and Quarantine, Beijing 100123, China.
Int J Environ Res Public Health. 2015 Dec 9;12(12):15673-82. doi: 10.3390/ijerph121215012.
This study aimed to explore the effects of perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) on apoptosis and cell cycle in a zebrafish (Danio rerio) liver cell line (ZFL). Treatment groups included a control group, PFOA-IC(50), PFOA-IC(80), PFOS-IC50 and PFOS-IC(80) groups. IC(50) and IC(80) concentrations were identified by cellular modeling and MTT assays. mRNA levels of p53, Bcl-2, Bax, Caspase-3 and NF-κB p65 were detected by qPCR. Cell apoptosis and cell cycle were detected by flow cytometry and the protein levels of p53, Bcl-2, Bax, Caspase-3 and NF-κB p65 were determined by western blotting. Both PFOA and PFOS inhibited the growth of zebrafish liver cells, and the inhibition rate of PFOS was higher than that of PFOA. Bcl-2 expression levels in the four groups were significantly higher than the control group and Bcl-2 increased significantly in the PFOA-IC(80) group. However, the expression levels of Bax in the four treatment groups were higher than the control group. The percentage of cell apoptosis increased significantly with the treatment of PFOA and PFOS (p < 0.05). Cell cycle and cell proliferation were blocked in both the PFOA-IC(80) and PFOS-IC(80) groups, indicating that PFOA-IC(80) and PFOS-IC(50) enhanced apoptosis in ZFL cells.
本研究旨在探讨全氟辛酸（PFOA）和全氟辛烷磺酸（PFOS）对斑马鱼（Danio rerio）肝细胞系（ZFL）凋亡和细胞周期的影响。处理组包括对照组、PFOA-IC(50)组、PFOA-IC(80)组、PFOS-IC50组和PFOS-IC(80)组。通过细胞建模和MTT试验确定IC(50)和IC(80)浓度。采用qPCR检测p53、Bcl-2、Bax、Caspase-3和NF-κB p65的mRNA水平。通过流式细胞术检测细胞凋亡和细胞周期，采用蛋白质印迹法测定p53、Bcl-2、Bax、Caspase-3和NF-κB p65的蛋白质水平。PFOA和PFOS均抑制斑马鱼肝细胞生长，且PFOS的抑制率高于PFOA。四组中Bcl-2表达水平均显著高于对照组，且PFOA-IC(80)组Bcl-2显著升高。然而，四个处理组中Bax的表达水平均高于对照组。PFOA和PFOS处理后细胞凋亡百分比显著增加（p < 0.05）。PFOA-IC(80)组和PFOS-IC(80)组的细胞周期和细胞增殖均受到阻滞，表明PFOA-IC(80)和PFOS-IC(50)增强了ZFL细胞的凋亡。
