三丁基锡引起人羊膜细胞氧化损伤及DNA损伤的研究

背景与目的： 研究三丁基锡 (tributyltin,TBT)对人羊膜细胞FL (human amnion cells) 氧化损伤和DNA损伤的诱导作用。 材料与方法： 将不同浓度TBT (0、2、4、6、8、10 μmol/L)，分别对FL染毒2 h和4 h，各染毒组同时设不加TBT的对照组，染毒后分别用MTT法检测TBT对FL细胞增殖率的影响，用DCFH_DA法检测FL细胞活性氧自由基 (ROS)水平，用彗星实验检测TBT对FL细胞DNA的损伤。 结果： TBT对FL细胞染毒4 h时，其2、8、10 μmol/L浓度组的细胞增殖率较对照组显著下降(P<0.05，P<0.01，P<0.001)，且随TBT浓度升高而增殖率呈下降的趋势。TBT 3、4 μmol/L染毒组FL细胞的ROS水平较对照组升高，且4 μmol/L染毒组与对照组比较差异具有统计学意义(P<0.05)。在TBT 2、3、4 μmol/L染毒组随着TBT浓度的升高，FL细胞核尾长、尾相均显著升高(P均<0.05)。 结论： TBT可引起FL细胞的氧化损伤及DNA损伤。

Study of Oxidative Damage and DNA Damage in FL Cells Induced by Tributyltin

BACKGROUND AND AIM: The study was undertaken to assess the oxidative stress and DNA damage in human amnion cells induced by TBT. MATERIALS AND METHODS: Cultured FL cells were exposed to different concentrations of TBT (0, 2, 4, 6, 8, 10 μmol/L) for different durations (2 h and 4 h). Cell proliferation rate was evaluated by MTT assay. After exposure to different concentrations of TBT (0,1,2,3,4 μmol/L) for 2 h,the ROS level and DNA damage of FL cells were measured by DCFH_DA method and the single cell gel electrophoresis method, respectively. RESULTS: After exposure to different concentrations of TBT for 4 h, the FL cell proliferation rates in the 2, 8, 10 μmol/L groups were significantly decreased as compared to control(P<0.05, P<0.01, P<0.001) in a dose_dependent manner. The ROS level was increased in the 3 and 4 μmol/L groups in the dose_dependent manner and the difference was significant at 4 μmol/L group compared to the control(P<0.05). Meanwhile, TBT exposure led to the interrelated increase of nucleus tail length and tail moment in a dose－dependent manner and the differences were significant at 2, 3 and 4 μmol/L group compared to the control(P<0.05). CONCLUSION: TBT exposure could cause oxidative stress and DNA damage to FL cells.