摘要
[目的]研究乙酸铅染毒人外周血淋巴细胞致氧化应激与DNA氧化损伤情况。[方法]分别用浓度为0μmol/L、20μmol/L、40μmol/L和80μmol/L的乙酸铅染毒人外周血淋巴细胞6 h、12 h、24 h,应用2’,7’-二氯二氢荧光素二乙酸酯(DCFH-DA)染色分析和流式细胞仪检测染毒后细胞内活性氧类(ROS)水平,高度水溶性四唑盐(WST-1)法检测染毒后细胞内总超氧化物歧化酶(T-SOD)活性,酶联免疫吸附测定法(ELISA)试剂盒检测染毒后细胞8-羟基脱氧鸟苷(8-OHdG)水平,并对ROS、T-SOD、8-OHdG三指标间的关系进行相关性分析。[结果]乙酸铅染毒6 h后,各染毒组人外周血淋巴细胞ROS水平均高于对照组,差异有统计学意义(P <0.01);20μmol/L染毒组人外周血淋巴细胞T-SOD和8-OHdG水平与对照组相比,差异无统计学意义(P> 0.05),而40μmol/L和80μmol/L染毒组人外周血淋巴细胞T-SOD和8-OHdG水平均高于对照组,差异有统计学意义(P <0.01)。乙酸铅染毒12 h和24 h后,各染毒组人外周血淋巴细胞ROS、T-SOD和8-OHdG水平均高于对照组,差异有统计学意义(P <0.01)。相关性分析显示,细胞内ROS含量与T-SOD活性呈负相关(r_(6 h)=-0.865、r_(12 h)=-0.890、r_(24 h)=-0.801,P <0.01),与8-OHdG含量呈正相关(r_(6 h)=0.840、r_(12 h)=0.829、r_(24 h)=0.866,P <0.01)。[结论]乙酸铅染毒人外周血淋巴细胞后会诱导细胞ROS生成,抑制细胞内抗氧化酶SOD活性,造成人外周血淋巴细胞氧化应激状态增强和DNA氧化性损伤。
[Objective] To observe oxidative stress and DNA oxidative damage of human peripheral blood lymphocytes in vitro induced by lead acetate. [Methods] Human peripheral blood lymphocytes were exposed to lead acetate in vitro at concentrations of 0,20,40,80 μmol/L for 6,12,24 h,respectively. Then the intracellular reactive oxygen species（ROS）levels were detected by DCFH-DA staining and flow cytometry. Meanwhile,the activity of total superoxide dismutase（T-SOD）was detected by WST-1 method,and the 8-OHdG level was detected by ELISA Kit. The correlation analysis was carried out among ROS,T-SOD and8-OHdG. [Results] After exposed to lead acetate in vitro for 6 h,ROS levels of human peripheral blood lymphocytes in each dose group were significantly higher than those in control group（P〈 0.01）. There was no significant difference of T-SOD and 8-OHdG levels in 20 μmol/L group compared with those in control group（P 〉0.05）,however,T-SOD and 8-OHdG levels of peripheral blood lymphocytes in the 40 and 80 μmol/L groups were significantly higher than those in control group（P 〈0.01）.After exposed to lead acetate for 12 and 24 h,the levels of ROS,T-SOD and 8-OHdG of peripheral blood lymphocytes in each dose group were significantly higher than those in the control group（P 〈0.01）. There were significant negative correlations between ROS levels and T-SOD levels（r6 h=-0.865,r12 h=-0.890,r24 h=-0.801,P〈 0.01）,and there were significant positive correlations between ROS levels and 8-OHdG levels（r6 h= 0.840,r12 h= 0.829,r24 h= 0.866,P〈 0.01）. [Conclusion] The study findings indicate that lead acetate could induce ROS production and inhibit the activity of antioxidant enzyme SOD,and increase oxidative stress state and DNA oxidative damage of human peripheral blood lymphocytes in vitro.
作者
刘祥铨
吴京颖
刘雯
施文华
吴小南
刘合焜
LIU Xiangquan1,WU Jingying1,LIU Wen1,SHI Wenhua1,WU Xiaonan2,LIU Hekun2(1. Fuzhou Center for Disease Control and Prevention, Fuzhou , Fujian 350004, China;2. Fujian Medical University, Fuzhou , Fujian 350108, China)
出处
《职业卫生与应急救援》
2018年第5期381-384,共4页
Occupational Health and Emergency Rescue
基金
福建省卫生系统中青年骨干人才培养项目资助(2014-ZQN-JC-35)
