双酚A及纳米二氧化钛对人胚肝L-02细胞内活性氧含量及DNA损伤的联合作用

Combined Effects of Bisphenoi A and Nano-TiO_2 on ROS and DNA Damage Level in Human Embryo Liver L-02 Cells

[目的]观察双酚A(BPA)、纳米二氧化钛(nano-TiO_2)单独及联合作用对人胚肝L-02细胞内活性氧(reactive oxygen species,ROS)含量及DNA损伤的影响。[方法]生长良好的L-02细胞,分别单独加入0.1、1、10μmol/L的BPA,1、5、10 mg/L的nano-TiO_2,以及上述各浓度的BPA和nano-TiO_2混合染毒24 h;采用流式细胞仪检测各组L-02细胞内ROS含量,采用彗星试验观察各组DNA断裂损伤程度。[结果]10μmol/L BPA单独染毒组,1mg/L nano-TiO_2与1、10μmol/L BPA,以及5、10 mg/L的nano-TiO_2分别与0.1、1、10μmol/L BPA联合作用于L-02细胞后,均可引起ROS含量升高,DNA断裂损伤加重,与阴性对照组比较差异有统计学意义(P<0.001);而nano-TiO_2单独染毒组对于L-02细胞ROS的升高、DNA损伤作用与阴性对照组比较差异均无统计学意义(P>0.05)。析因分析结果显示两种受试物混合染毒存在协同作用。[结论]BPA和nano-TiO_2联合作用可增加各自对L-02细胞内ROS水平和DNA损伤断裂水平的影响,对细胞的损伤作用增大。

[ Objective ] To investigate the effects of Bisphenol A （ BPA ）, nano-TiO2, and BPA＋nano-TiO2 on ROS and DNA damage level in haman embryo liver L-02 cells. [ Methods ] L-02 cells were exposed to single BPA（0.1, 1, 10 μmol/L）, single nano-TiO2 （ 1, 5, 10 mg/L ）, and combination of BPA in different concentrations with nano-TiO2 respectively. Reactive oxygen species （ ROS ） level was determined by flow eytometry, and DNA damage was measured by comet assay. [ Results ] Compared with negative control group, increased ROS and DNA damage level were found in groups of 10 oanol/LBPA; 1 mg/L nano-TiO2 ＋ 1, 10 μmol/L BPA ; 5, 10 mg/L nano-TiO2 ＋ 0.1, 1, 10 μmol/L BPA, respectively.＇ No significant effects on ROS and DNA damage were observed in single nano-TiO2 groups （P 〉 0.05 ）. Coefficient effects on ROS and DNA damage were observed in the combined groups. [ Conclusion ] Combined usage of BPA and nano-TiO2 enhanced their capability in ROS induction and DNA damage effects on L-02 cells.