活性氧在邻苯二甲酸单(2-乙基己基)酯所致心肌细胞DNA损伤中的作用  

Effect of reactive oxidative species on MEHP-induced DNA damage in human cardiac cells

作　　者：王泽泽 陈晋波[3] 刘君瑶 杨璟 李龙飞 孙鹤鸣 张一欣 郭会彩[1,4] WANG Ze-ze;CHEN Jin-bo;LIU Jun-yao;YANG Jing;LI Long-fei;SUN He-ming;ZHANG Yi-xin;GUO Hui-cai(Department of Toxicology,School of Public Health,Hebei Medical University,Shijiazhuang Hebei 050017,China;Department of Tropical Medicine,College of Military Preventive Medicine,Army Medical University,Chongqing 400000,China;The Second Hospital of Shijiazhuang City,Hebei Province,Shijiazhuang Hebei 050051,China;Hebei Key Laboratory of Environment and Human Health,Shijiazhuang Hebei 050017,China)

机构地区：[1]河北医科大学公共卫生学院毒理学教研室,河北石家庄050017 [2]陆军军医大学军事预防医学系热带医学教研室,重庆400000 [3]河北省石家庄市第二医院,河北石家庄050051 [4]河北省环境与人群健康重点实验室,河北石家庄050017

出　　处：《毒理学杂志》2022年第1期10-14,共5页Journal of Toxicology

基　　金：国家自然科学基金(21976050);石家庄市科学技术研究与发展计划项目(191460933)。

摘　　要：目的探讨邻苯二甲酸单(2-乙基己基)酯(MEHP)对人源心肌AC16细胞DNA的损伤作用及其可能机制。方法以人源心肌AC16细胞为研究对象,分为对照组,40、60、80、100和120μmol/L MEHP组。处理细胞24 h后采用DCFH-DA探针标记测定AC16细胞内活性氧(ROS)水平;单细胞凝胶电泳实验测定细胞DNA损伤情况;利用抗氧化剂N-乙酰-L-半胱氨酸(NAC)抑制AC16细胞内ROS生成,以观察ROS在MEHP所致AC16细胞DNA损伤中的作用。结果与对照组相比,AC16细胞经MEHP处理后,ROS水平随着MEHP浓度的增加而明显升高(P<0.05);对照组心肌细胞核呈圆形荧光团,强度均匀,边缘光滑,大小较一致,无明显拖尾。不同浓度MEHP处理后,各组心肌细胞出现彗星拖尾现象,且具有MEHP浓度依赖性。与对照组OTM值和TD值相比,MEHP组OTM值和TD值明显升高,且有统计学意义(P<0.05);预先给予NAC可明显降低MEHP所致的细胞ROS水平升高。与相应的MEHP组相比,NAC+MEHP组OTM值及TD值明显降低,且差异有统计学意义(P<0.05)。结论MEHP可引起人源心肌AC16细胞DNA损伤,机制可能与ROS生成有关。Objective To investigate the role of reactive oxygen species(ROS)in MEHP-induced DNA damage of human cardiac AC16 cells.Methods The AC16 cells were treated with MEHP(40,60,80,100 and 120μmol/L)or solvent control for 24 h.After treatment,the reactive oxygen species(ROS)content was assessed by fluorescent probes DCFH-DA,while DNA damage was evaluated with comet assay.Furthermore,antioxidant N-acetylcysteine(NAC)was added to evaluate the effect of ROS in MEHP-induced DNA damage of AC16 cells.Results Compared to the control group,there was a concentration-dependent increase in ROS accumulation of AC16 cells upon MEHP treatment(P<0.05).In the control group,there showed yellow-round nuclei of AC16 cells with homogenous distribution and consistent size,the edges were smooth without comet tail.MEHP treatment induced significant DNA damage in AC16 cells with a dose-dependent manner,which displayed as the enlongated comet tail.The OTM values as well as percentage of tail DNA contents(the TD values)in MEHP-treatmented AC16 cells were markedly higher than that of the cells in the control group(P<0.05).Pre-treatment with NAC significantly attenuated the enhanced ROS content induced by MEHP.Besides,in the NAC addition groups,the OTM and TD values decreased significantly in the NAC+MEHP group compared with the MEHP-treatment group(P<0.05).Conclusion MEHP may induce DNA damage in AC16 cells,and oxidative stress might be involved in the underlying mechanisms.

关 键 词：邻苯二甲酸单(2-乙基己基)酯 心肌细胞 活性氧 DNA损伤 

分 类 号：R114[医药卫生—卫生毒理学] R99[医药卫生—公共卫生与预防医学]