1,4-萘醌老化黑碳对人支气管上皮细胞活性氧和DNA链断裂的影响  

Effect of 1,4-naphthoquinone aged black carbon on reactive oxygen species and DNA strand breaks in human bronchial epithelial cells

作　　者：张文晓[1] 尚静[2] 郜鑫[1] 栾先国 李苹[1] 王天晶[1] 胡贵平[1] 朱彤[2] 贾光[1] 

机构地区：[1]北京大学公共卫生学院劳动卫生与环境卫生学系,北京100191 [2]北京大学环境科学与工程学院,北京100871

出　　处：《北京大学学报（医学版）》2015年第4期690-696,共7页Journal of Peking University:Health Sciences

基　　金：国家自然科学基金(21190051)资助~~

摘　　要：目的:研究1,4-萘醌老化黑碳(1,4-naphthoquinone aged black carbon,BC/1,4-NQ)对人支气管上皮细胞(16HBE)活性氧水平和DNA链断裂的影响。方法:分别用BC/1,4-NQ及相对应质量浓度的BC和1,4-NQ(BC/1,4-NQ的质量浓度分别为:10.0/0.2、20.0/0.4、40.0/0.8、80.0/1.6、160.0/3.2 mg/L,BC的质量浓度分别为:10.0、20.0、40.0、80.0、160.0 mg/L,1,4-NQ的质量浓度分别为:0.2、0.4、0.8、1.6、3.2 mg/L)染毒16HBE细胞24、48、72 h,通过cell counting kit-8(CCK-8)法检测细胞毒性。再以不同剂量的BC/1,4-NQ(20.0/0.4、40.0/0.8、80.0/1.6 mg/L)、BC(20.0、40.0、80.0 mg/L)、1,4-NQ(0.4、0.8、1.6 mg/L)染毒16HBE细胞24 h,采用2',7'-二氯荧光黄双乙酸盐探针(DCFH-DA)检测细胞内活性氧(reactive oxygen species,ROS)水平,通过单细胞凝胶电泳实验,以Olive尾距为指标,评价DNA链断裂遗传毒性。结果:除10.0/0.2 mg/L剂量染毒24 h后BC/1,4-NQ未见明显的细胞毒性,在各染毒时间点、各剂量BC/1,4-NQ的细胞存活率均显著低于对照组(P<0.05),并表现出一定的剂量依赖效应。当BC/1,4-NQ≥80.0/1.6 mg/L时,BC/1,4-NQ的细胞存活率要低于BC单独处理组、高于1,4-NQ处理组(P<0.05)。此外,各剂量BC/1,4-NQ均可引起16HBE胞内ROS含量及Olive尾距增加,并显著大于对照组(P<0.05),表现出一定的剂量依赖效应;当BC/1,4-NQ为80.0/1.6 mg/L时,BC/1,4-NQ的胞内ROS含量和Olive尾距显著大于BC单独处理组、小于1,4-NQ处理组(P<0.05)。结论:以BC/1,4-NQ处理16HBE细胞,可诱导细胞内ROS水平升高,并产生细胞毒性和遗传毒性,且在较高剂量下,BC/1,4-NQ的细胞毒性、胞内ROS水平和遗传毒性要高于BC单独处理组,低于1,4-NQ单独处理组。Objective： To study the effect of 1,4-naphthoquinone aged black carbon（ BC /1,4-NQ） on reactive oxygen species and DNA strand breaks in human bronchial epithelial cells（ 16HBE）. Methods：In the study,16 HBE cells were exposed to BC /1,4-NQ,BC and 1,4-NQ at the concentrations of BC /1,4-NQ（ 10. 0 /0. 2,20. 0 /0. 4,40. 0 /0. 8,80. 0 /1. 6,160. 0 /3. 2 mg / L）,BC（ 10. 0,20. 0,40. 0,80. 0,160. 0 mg / L）,1,4-NQ（ 0. 2,0. 4,0. 8,1. 6,3. 2 mg / L） for 24,48,and 72 h,respectively.Cytotoxicity was detected by cell count kit 8（ CCK-8） at the end point. Then the 16 HBE cells were exposed to BC /1,4-NQ（ 20. 0 /0. 4,40. 0 /0. 8,80. 0 /1. 6 mg / L）,BC（ 20. 0,40. 0,80. 0 mg / L）,1,4-NQ（ 0. 4,0. 8,1. 6 mg / L） for 24 h. The reactive oxygen species（ ROS） generation was determined via flow cytometry with DCFH-DA probe. Single cell gel electrophoresis（ SCGE） assay was performed to evaluate genotoxicity by Olive tail moment（ OTM） value. Results： Except for the concentration of 10. 0 /0. 2 mg / L within the exposure time 24 h,the cell viabilities of BC /1,4-NQ were significantly lower than the control（ P〈0. 05） within the exposure time 24- 72 h,showing a dose-dependent cytotoxicity. Especially,BC /1,4-NQ showed greater cytotoxicity than BC single exposure,lower than 1,4-NQ at the concentration of BC /1,4-NQ≥80. 0 /1. 6 mg / L. BC /1,4-NQ also showed greater ROS generation and OTM value than the control within the exposure time 24 h at each concentration（ P〈0. 05）. Especially,the ROS generation and OTM value of BC /1,4-NQ were greater than BC single exposure,lower than 1,4-NQ at the concentration of 80. 0 /1. 6 mg / L（ P〈0. 05）. Conclusion： BC /1,4-NQ can induce intracellular ROS generation,cytotoxicity and genotoxicity in 16 HBE cells. And at high concentration,the intracellular ROS level,cytotoxicity and genotoxicity induced by BC /1,4-NQ were greater than those by BC single exposure,but lower than those by 1,4-NQ.

关 键 词：煤烟 1 4-萘醌 支气管上皮细胞 DNA断裂 活性氧 

分 类 号：R122.7[医药卫生—环境卫生学]