纳米二氧化钛与醋酸铅联合诱导人胚肝细胞氧化应激作用  

Joint Oxidative Stress Induced by Nanometer Titanium Dioxide and Lead Acetate in Human Derived Fetal Hepatocytes

作　　者：杜海荣 朱晓玲 周宜开[3] 

机构地区：[1]深圳市光明新区疾病预防控制中心,广东深圳518106 [2]深圳市职业病防治院,广东深圳518000 [3]华中科技大学同济医学院公共卫生学院环境医学研究所,环境与健康教育部重点实验室,湖北武汉430030

出　　处：《环境与职业医学》2012年第9期552-555,共4页Journal of Environmental and Occupational Medicine

基　　金：国家自然科学基金资助项目(编号:20777025);深圳市科技计划项目(医疗卫生类)赞助(编号:201103217)

摘　　要：[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 ] To study the joint effect on reactive oxygen species （ROS） generation, cell viabil.ity, and oxidative stress induced by nanometer titanium dioxide （TiO2） and lead acetate （PbAc） in human derived fetal hepatocytes （L-02）. [ Methods ] The experimental groups were treated with PbAc （1.000mg/L）, TiO2 （10.000, 1.000, 0.100, 0.010, 0.001 rag/L）, or all concentration combinations of PbAc and TiO2 for 24 h. The L-02 cells were also cultured with 0.1% dimethyl sulfoxide （negative control） and 30μmol/L H2O2 （positive control）. Cell toxicity and ROS generation were determined using methyhhiazoltetrazolium assay and flow cytometry respectively. The levels of glutathione （GSI-I） and activities of superoxide dismutase （SOD） were used to determine intracellular antioxidant levels. [ Results ] When treated with 10.000mg/L TiO2, the cell viability was decreased significantly＇compared with the negative control, the PbAc, and the other single TiO2 dosage groups （P〈0.05）. When interfered with the combination of 1.000 mg/L PbAc ＋ 10.000 mg/L TiO2, the cell viability was reduced significantly compared with the negative control, the PbAc, and the other combination groups （P 〈 0.05）. The cytotoxicity was notably higher in the combination group of 1.000 mg/L PbAc ＋1.000 or 0.100 mg/L TiO2 than in the negative controls （P 〈 0.05）. There were significant increases of the ROS levels in various combination groups in comparison with the negative controls and the PbAe group （P 〈 0.05）. The GSH levels were significantly increased after combined treatment of 1.000mg/L PbAc ＋ 1.000, 0.100 or 0.010mg/L TiO2 （P〈O.05）, and the SOD levels were notably raised after 1.000mg/L PbAc ＋ 0.100 or 0.010mg/L TiO2 combined treatment, incomparison with the negative controls （P 〈 0.05）. [ Conclusion ] In these study settings, co-exposure to low doses of TiO2 and PbAc significantly induces elevated ROS levels in L-02, and subsequently elevation of GSH and SOD leve

关 键 词：纳米二氧化钛 醋酸铅 联合作用 氧化应激 

分 类 号：R114[医药卫生—卫生毒理学]