无机砷对Chang肝细胞活性氧的诱导和核转录因子表达的影响

Induction of ROS and Expression of Transcription Factor Nrf 2 in Chang Liver Cells Induced by Inorganic Arsenic

目的研究亚砷酸钠(NaAsO_2)对Chang肝细胞中活性氧(ROS)的诱导以及对核转录因子Nrf2的活化作用。方法体外实验采用0～10μmol/LNaAsO_2溶液对Chang肝细胞染毒2、6、12和24 h。分别用流式细胞仪和免疫印记实验(Western blot)技术检测细胞内ROS含量和Nrf2蛋白的表达。结果2.5μmol/lNaAsO_2溶液染毒6 h、5.0μmol/L NaAsO_2溶液染毒2、6、12、24h的平均荧光强度的相对比值均高于对照组,差异有统计学意义(P<0.05或P<0.01);且ROS的产生量随着NaAsO_2溶液浓度的升高而增多。5.0μmol/L NaAsO_2溶液染毒12 h、5.0μmol/L NaAsO_2溶液染毒2、6、12 h的核转录因子Nrf2蛋白表达均强于对照组,差异有统计学意义(P<0.01);且核转录因子Nrf2蛋白表达的活化具有时间相关性,即在NaAsO_2溶液染毒12 h时核转录因子Nrf2蛋白表达的活化出现高峰,而后逐渐降低至正常水平。结论无机砷能够诱导Chang肝细胞内ROS的生成和增强核转录因子Nrf2蛋白的表达。

Objective To study whether sodium arsenite （NaAsO2） can induce the production of reactive oxygen species （ROS） and activate the expression of transcription factor NF-E2-related factor （Nrf 2）. Methods Chang liver cell were treated with NaAsO2at doses of 0 umol/L to 10 umol/L for 2, 6, 12 and 24 h, respectively. ROS and expression of Nrf 2 protein in ceils exposed to NaAsO2 were measured with flow cytometry （FACS） and Western blot, respectively. Results The relative average fluorescence intensity in the cells treated with 2.5 umol/L NaAsO2 for 6 h and 5.0 umol/L NaAsO2 for 2, 6, 12 and 24 h respectively increased significant compared with the controls （P〈0.01）. The expression of Nrf 2 reached the peak at 12 h after NaAsO2 treated, while degraded to normal level gradually. Conclusion NaAsO2 can induce the production of ROS and up-regulate the expression of Nrf 2.