环境与职业医学2024，Vol.41Issue(6)：681-686,6.DOI:10.11836/JEOM23408
天然除虫菊素诱导人肝细胞DNA氧化损伤研究
DNA oxidative damage induced by natural pyrethrins in human liver cells
摘要
Abstract
[Background]Natural pyrethrins have long been widely used in the fields of environmental and household hygiene.Studies have reported that natural pyrethrins have potential liver toxicity,but their specific mechanisms are still unclear yet. [Objective]To explore the effect of natural pyrethrins on DNA damage in human liver cells. [Methods]This study used human liver cell QSG7701 as an in vitro testing model.After exposure to DMSO and a series of concentrations of natural pyrethrins(5,10,20,and 40 μg·mL-1)for 6 and 24 h,reactive oxygen species(ROS)was detected by fluorescence microscopy using a fluorescence probe,thiobarbituric acid reactive substance(TBARS)by colorimetric method using a microplate reader,DNA damage by comet assay through observing DNA fragment migration under microscope,and phospho H2AX(γH2AX)and 8-oxoguanine(8-oxoG)by immunofluorescence assay using a laser confocal microscope. [Results]As the exposure concentration of natural pyrethrins increased,the fluorescence intensity of ROS significantly increased in a concentration-dependent manner.The differences in ROS between the 10 μg·mL-1 and above groups and the control group were statistically significant(P<0.01),and the ROS levels in the 20 μg·mL-1 and 40 μg·mL-1 treatment groups were 2.17 and 3.05 times higher than that in the control group respectively.The TBARS level increased in a concentration-dependent manner in natural pyrethrins treated cells(P<0.01),and the levels in the 20 μg·mL-1 and 40 μg·mL-1 treatment groups were 2.46 and 3.01 times higher than that in the control group respectively.The results of comet assay showed trailing formation of cellular DNA in each dose group;as the exposure concentration of natural pyrethrins increased,indicators such as tail DNA content(TDNA%),tail length(TL),tail moment(TM),and Olive tail moment(OTM)increased in a concentration-dependent manner.Compared with the control group,the differences in the indicators between the 20 μg·mL-1 and above groups and the control group were statistically significant(P<0.01),especially in the 40 μg·mL-1 treatment groups,where TDNA%,TL,TM,and OTM were(46.92±3.52)%,(64.67±4.16)μm,30.96±2.94,and 22.64±3.89,respectively.The cellular im-munofluorescence results showed that natural pyrethrins induced the formation of γH2AX and 8-oxoG,the fluorescence intensities of γH2AX and 8-oxoG increased in a concentration-dependent manner,and the differences between the 10 μg·mL-1 and above groups and the control group were statistically significant(P<0.01). [Conclusion]Natural pyrethrins could induce DNA damage in human liver cells,and ROS-mediated oxidative stress may play an important role in its liver cell genotoxicity.关键词
天然除虫菊素/人肝细胞/活性氧/氧化应激/DNA损伤/遗传毒性Key words
natural pyrethrin/hepatocyte/reactive oxygen species/oxidative stress/DNA damage/genotoxicity分类
医药卫生引用本文复制引用
杨韵,应梦超,孙静秋,沙毅杰,洪新宇,肖萍,陶功华..天然除虫菊素诱导人肝细胞DNA氧化损伤研究[J].环境与职业医学,2024,41(6):681-686,6.基金项目
上海市青年科技英才扬帆计划项目(21YF1439100) （21YF1439100）
上海市公共卫生体系建设三年行动计划重点学科建设项目(GWVI-11.1-41) （GWVI-11.1-41）
国家环境保护重点实验室新兴污染物环境健康影响评价专项基金项目(SEPKL-EHI-AEC-202204) （SEPKL-EHI-AEC-202204）
上海市卫生健康委员会卫生行业临床研究专项面上项目(202140489) （202140489）
