This study investigated the conjoined cellular oxidative damage of human embryo kidney 293T（HEK293T） cells induced by cadmium chloride（CdCl2） and nanometer titanium dioxide（nano-TiO2）.RT-PCR technique was used t...This study investigated the conjoined cellular oxidative damage of human embryo kidney 293T（HEK293T） cells induced by cadmium chloride（CdCl2） and nanometer titanium dioxide（nano-TiO2）.RT-PCR technique was used to detect the expressions of Heme oxygenase-1（HO-1） and 8-oxoguanine DNA glycosylase（OGG1）.The activities of superoxide dismutase（SOD） and catalase enzyme（CAT） and concentrations of reactive oxygen species（ROS） and maldondialdehyde（MDA） were measured by different approaches.The results showed that CdCl2 and nano-TiO2 at a low concen-tration of 0.75 total toxic unit（TU） exerted an additive effects on HO-1 gene expression,CAT activities and MDA concentrations.When the total TU was increased to 1 or 1.25 TU,the interaction was syner-getic.Moreover,the mixture with high proportion of CdCl2 produced an additive effect on the OGG1 gene expression,and the interaction was changed to be synergetic when the concentration of CdCl2 was lower than or equal to that of nano-TiO2.Synergetic effects of CdCl2 and nano-TiO2 on cellular oxida-tive damage of HEK293T cells were found as indicated by the changes in the SOD activities and ROS concentrations.It was concluded that CdCl2 and nano-TiO2 exerts synergistic effects on the cellular oxidative damage of HEK293T cells,and the sensitivity of these indicators of oxidative damage varies with the proportion of CdCl2 and nano-TiO2 in the mixture.展开更多
目的探讨镉暴露对HepG2细胞转录因子NF-E2相关因子2(NRF2)信号通路的影响。方法采用甲臢比色法测定CdCl_2(0、1、2.5、5、10、25、50、100、200μmol/L)处理24 h后,HepG2细胞活力变化;应用蛋白免疫印迹法检测CdCl_2(1、2、5、10、20μmo...目的探讨镉暴露对HepG2细胞转录因子NF-E2相关因子2(NRF2)信号通路的影响。方法采用甲臢比色法测定CdCl_2(0、1、2.5、5、10、25、50、100、200μmol/L)处理24 h后,HepG2细胞活力变化;应用蛋白免疫印迹法检测CdCl_2(1、2、5、10、20μmol/L)处理细胞6 h后,NRF2蛋白水平;采用RT-q PCR方法检测10μmol/L CdCl_2处理细胞2、4、6、12、24 h后,GCLC、GCLM、HO1和AKR1C1 m RNA水平变化,检测CdCl_2(1、2、5、10、20μmol/L)处理细胞6 h后,GCLC、GCLM、HO1和AKR1C1 m RNA水平变化。结果 HepG2细胞活力随镉处理剂量升高而降低(P<0.05);与对照组(0.60±0.01)比较,1、2、5、10、20μmol/L镉处理组HepG2细胞NRF2蛋白表达水平[分别为(0.65±0.01)、(1.37±0.04)、(1.94±0.05)、(2.24±0.07)、(2.22±0.05)]均明显升高(P<0.05);与对照组比较,镉处理6 h时,HepG2细胞内GCLC、GCLM、HO1和AKR1C1 m RNA水平[分别为(45.76±7.04)、(114.21±5.23)、(59.52±1.50)、(674.13±27.12)]明显升高(P<0.05);与对照组比较,5μmol/L镉处理组HepG2细胞内GCLC和GCLM m RNA水平[分别为(24.77±2.16)、(29.93±0.67)]升高,2μmol/L镉处理组HepG2细胞内HO1和AKR1C1 m RNA水平[分别(28.55±2.02)、(186.32±12.63)]升高(P<0.05)。结论镉暴露能激活HepG2细胞系中NRF2信号通路。展开更多
基金the National Investigation Projects of Soil Pollution supported by a grant from Ministry of Environmental Protection of the People's Republic of China
文摘This study investigated the conjoined cellular oxidative damage of human embryo kidney 293T（HEK293T） cells induced by cadmium chloride（CdCl2） and nanometer titanium dioxide（nano-TiO2）.RT-PCR technique was used to detect the expressions of Heme oxygenase-1（HO-1） and 8-oxoguanine DNA glycosylase（OGG1）.The activities of superoxide dismutase（SOD） and catalase enzyme（CAT） and concentrations of reactive oxygen species（ROS） and maldondialdehyde（MDA） were measured by different approaches.The results showed that CdCl2 and nano-TiO2 at a low concen-tration of 0.75 total toxic unit（TU） exerted an additive effects on HO-1 gene expression,CAT activities and MDA concentrations.When the total TU was increased to 1 or 1.25 TU,the interaction was syner-getic.Moreover,the mixture with high proportion of CdCl2 produced an additive effect on the OGG1 gene expression,and the interaction was changed to be synergetic when the concentration of CdCl2 was lower than or equal to that of nano-TiO2.Synergetic effects of CdCl2 and nano-TiO2 on cellular oxida-tive damage of HEK293T cells were found as indicated by the changes in the SOD activities and ROS concentrations.It was concluded that CdCl2 and nano-TiO2 exerts synergistic effects on the cellular oxidative damage of HEK293T cells,and the sensitivity of these indicators of oxidative damage varies with the proportion of CdCl2 and nano-TiO2 in the mixture.
文摘目的探讨镉暴露对HepG2细胞转录因子NF-E2相关因子2(NRF2)信号通路的影响。方法采用甲臢比色法测定CdCl_2(0、1、2.5、5、10、25、50、100、200μmol/L)处理24 h后,HepG2细胞活力变化;应用蛋白免疫印迹法检测CdCl_2(1、2、5、10、20μmol/L)处理细胞6 h后,NRF2蛋白水平;采用RT-q PCR方法检测10μmol/L CdCl_2处理细胞2、4、6、12、24 h后,GCLC、GCLM、HO1和AKR1C1 m RNA水平变化,检测CdCl_2(1、2、5、10、20μmol/L)处理细胞6 h后,GCLC、GCLM、HO1和AKR1C1 m RNA水平变化。结果 HepG2细胞活力随镉处理剂量升高而降低(P<0.05);与对照组(0.60±0.01)比较,1、2、5、10、20μmol/L镉处理组HepG2细胞NRF2蛋白表达水平[分别为(0.65±0.01)、(1.37±0.04)、(1.94±0.05)、(2.24±0.07)、(2.22±0.05)]均明显升高(P<0.05);与对照组比较,镉处理6 h时,HepG2细胞内GCLC、GCLM、HO1和AKR1C1 m RNA水平[分别为(45.76±7.04)、(114.21±5.23)、(59.52±1.50)、(674.13±27.12)]明显升高(P<0.05);与对照组比较,5μmol/L镉处理组HepG2细胞内GCLC和GCLM m RNA水平[分别为(24.77±2.16)、(29.93±0.67)]升高,2μmol/L镉处理组HepG2细胞内HO1和AKR1C1 m RNA水平[分别(28.55±2.02)、(186.32±12.63)]升高(P<0.05)。结论镉暴露能激活HepG2细胞系中NRF2信号通路。
