论著

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

  • 张文晓 ,
  • 尚静 ,
  • 郜鑫 ,
  • 栾先国 ,
  • 李苹 ,
  • 王天晶 ,
  • 胡贵平 ,
  • 朱彤 ,
  • 贾光
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  • (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)

网络出版日期: 2015-08-18

基金资助

国家自然科学基金(21190051)资助

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

  • ZHANG Wen-Xiao ,
  • SHANG Jing ,
  • GAO Xin ,
  • LUAN Xian-Guo ,
  • LI Ping ,
  • WANG Tian-Jing ,
  • HU Gui-Ping ,
  • ZHU Tong ,
  • JIA Guang
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  • (1. Department of Occupational and Environmental Health Sciences, Peking University School of Public Health, Beijing 100191, China; 2. College of Environmental Sciences and Engineering, Peking University, Beijing 100871, China)

Online published: 2015-08-18

Supported by

Supported by the National Natural Science Foundation of China (21190051)

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。

本文引用格式

张文晓 , 尚静 , 郜鑫 , 栾先国 , 李苹 , 王天晶 , 胡贵平 , 朱彤 , 贾光 . 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[J]. 北京大学学报(医学版), 2015 , 47(4) : 690 -696 . DOI: 10.3969/j.issn.1671-167X.2015.04.028

Abstract

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, 16HBE 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 16HBE 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 16HBE 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.

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