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[目的]研究纳米二氧化钛(TiO2)与醋酸铅(PbAc)联合作用于人胚肝细胞(L-02),对细胞内活性氧(ROS)、氧化应激作用及细胞活性的影响。[方法]以1.000mg/L PbAc和10.000、1.000、0.100、0.010、0.001mg/L TiO2单独以及1.000mg/L PbAc及前述浓度TiO2混合处理L-02细胞24h,以体积分数为0.1%二甲基亚砜为阴性对照,30μmol/L H2O2为阳性对照。用噻唑蓝法检测细胞毒性,采用流式细胞术检测胞内ROS水平,检测谷胱甘肽(GSH)与超氧化物歧化酶(SOD)以评价细胞内抗氧化物质水平。[结果]与阴性对照、PbAc染毒组及其他浓度TiO2染毒组相比,10.000mg/L TiO2染毒组细胞活力明显降低(P<0.05)。与阴性对照、PbAc染毒组和其他浓度混合物染毒组相比,10.000mg/L混合物染毒组细胞活性明显降低(P<0.05);1.000、0.100mg/L混合物染毒组细胞活力也明显低于阴性对照组(P<0.05)。与阴性对照、PbAc染毒组相比,各浓度混合物染毒组细胞ROS水平均明显增加(P<0.05)。与阴性对照相比,1.000至0.010mg/L混合物染毒组细胞GSH水平均明显升高(P<0.05),0.100、0.010mg/L混合物染毒组细胞SOD活性也明显升高(P<0.05)。[结论]本研究条件下,低剂量纳米TiO2和PbAc共同作用于L-02,可增加活性氧水平,同时诱导细胞增加GSH和SOD水平以自我保护;随着染毒剂量升高,ROS水平明显增加,抗氧化能力下降,导致细胞活力下降。
[Objective] The research aimed to study the effects of titanium dioxide (TiO2) and lead acetate (PbAc) on human embryonic hepatocytes (L-02) and their effects on intracellular reactive oxygen species (ROS), oxidative stress and cell activity. [Method] L-02 cells were treated with 1.000mg / L PbAc and 10.000,1.000,0.100,0.010,0.001mg / L TiO2 alone and 1.000mg / L PbAc and the aforementioned concentration of TiO2 for 24 hours. The volume fraction of 0.1% Sulfoxide as a negative control, 30μmol / L H2O2 as a positive control. Cytotoxicity was detected by thiazolyl blue staining. Intracellular ROS levels were measured by flow cytometry. Glutathione (GSH) and superoxide dismutase (SOD) were measured to evaluate intracellular antioxidant levels. [Result] Compared with the negative control, PbAc and other concentrations of TiO2, the viability of cells in the treatment of 10.000mg / L TiO2 was significantly decreased (P <0.05). Compared with the negative control, the activity of cells in the mixture of 10 000 mg / L PbAc and other concentrations of mixture was significantly decreased (P <0.05). The viability of cells in the mixture of 1 000 and 0.100 mg / L was also significantly Lower than the negative control group (P <0.05). Compared with the negative control and the PbAc-treated group, the ROS levels of the exposed groups were significantly increased (P <0.05). Compared with the negative control, GSH levels in cells exposed to mixtures of 1.000 and 0.010 mg / L were significantly increased (P <0.05), and SOD activities in cells treated with mixtures of 0.100 and 0.010 mg / L were also significantly increased (P <0.05 ). [Conclusion] Under the conditions of this study, low-dose nano-TiO2 and PbAc combined with L-02 could increase the level of reactive oxygen species and induce cells to increase GSH and SOD levels to protect themselves. With the increase of exposure dose, the level of ROS significantly increased Increase, decreased antioxidant capacity, resulting in decreased cell viability.