DEHP对土壤蚯蚓氧化胁迫及DNA损伤的研究
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摘要
土壤环境中的酞酸酯污染日益严重,为了探讨和分析典型酞酸酯邻苯二甲酸二(2-乙基己基)酯(Di(2-ethylhexyl)phthalate,DEHP)对土壤动物的生态毒理效应,以赤子爱胜蚓为指示生物,暴露于DEHP浓度为CK、0.1、1、10、50 mg kg~(-1)人工土壤中,并于染毒后的7、14、21、28d取样测定。通过蚯蚓体内的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物(POD)、谷胱甘肽转移酶(GST)等指标反映DEHP对蚯蚓的氧化激活程度,通过活性氧自由基(ROS)的含量反映DEHP对蚯蚓的细胞毒性,通过丙二醛(MDA)含量和Olive尾矩反映DEHP对蚯蚓的遗传毒性,从多个角度评价DEHP对蚯蚓的生态毒理效应。结果表明:(1)在DEHP的刺激下,各浓度组蚯蚓的SOD、CAT、POD、GST活性均呈激活状态,表明DEHP对蚯蚓的抗氧化酶有诱导作用;(2)DEHP影响蚯蚓的ROS含量,各染毒处理组与对照组相比均升高且差异性明显,表现出明显的剂量—效应关系;(3)对比各染毒处理组之间的数据,DEHP对蚯蚓MDA含量的影响无明显规律;(4)DEHP可使蚯蚓GST呈激活状态,表明中高浓度的DEHP对蚯蚓GST具有诱导作用;(5)DEHP能够引起蚯蚓体腔细胞DNA的损伤,且随着浓度的增加,Olive尾矩值随之增加,说明DNA损伤程度与DEHP浓度之间具有剂量-效应关系。从实验结果可以看出,DEHP可以对蚯蚓机体和DNA造成一定程度的损伤,表现出较强的生态毒理效应。
【Objective】Phthalate acid esters(PAEs),also known as phthalate esters or "plasticizer",are the reaction product of phthalic anhydride and alcohol.PAEs are widely used in the production of a huge number of products such as agricultural plastic film,toys,medical bags and hoses for transfusion,food packaging,cleansers,vinyl floor and wallpaper,and personal care products.Among them,Di(2-ethylhexyl)phthalate(DEHP)is one PAE,the highest in output and consumption and also a toxic organic compound highly hazardous to human health,disturbing endocrine systems and causing cancers,mutation,oxidative damage and genetic toxicity.As DEHP is weakly bonded to the plastic matrix by hydrogen bond and van der Waals force,it is easily detached and diffused into the air,water and soil environments.Therefore,its pollution and hazard to human health has aroused more and more attention from the people.So far studies have proven that DEHP can enter organisms via a variety of ways,causing poisonous damage to tissues and organs of the organisms.As DEHP is extensively used in agriculture and hence exists in large volumes in the soil system,it is necessary to investigate its ecological and toxicological effects,such as oxidative stress and DNA damage,on typical soil animals,in the hope that the study may serve as scientific basis for reasonable assessment of soil ecological risk of DEHP.【Method】To explore eco-toxicological effects of DEHP on soil animals,Earthworms(Eisenia foetida),used as bio-indicator,were exposed to DEHP varying in concentration(0,0.1,1,10 and 50 mg kg~(-1))in artificial soil.Earthworms were sampled after 7,14,21 and 28 days of exposure for analysis of superoxide dismutase(SOD),catalase(CAT),peroxidase(POD)and glutathione(GST),which are the indices reflecting the degree of DEHP activating oxidation of earthworms,reactive oxygen species(ROS),which is the indicator of cytotoxicity of DEHP to earthworms,and malondiadehyde(MDA)and olive tail moment,which are the indices reflecting genotoxicity of the substance to earthworms.【Result】(1)DEHP,above 0.1 mg kg~(-1) in concentration,significantly enhanced the activities of SOD,CAT,POD and GST in Eisenia foetida by a degree varying with the concentration of DEHP,which demonstrates that DEHP induces antioxidant enzymes in earthworms;(2)DEHP increased ROS concentration in earthworms as compared with control and the effect varied between treatments,displaying an apparent dose-effect relationship;(3)Comparison between the treatments shows that DEHP did have some effect on content of MDA,but the effect did not follow any specific rule and was the most obvious in earthworms exposed to DEHP,10 mg kg~(-1) and 50 mg kg~(-1) in concentration;(4)In the presence of DEHP.GST in the earthworms was apparently activated,except for the treatment of 0.1 mg kg~(-1),where the earthworms did not have much difference from those in control.The observation demonstrates that DEHP,higher than 0.1 mg kg~(-1) in concentration induces GST in the earthworms;(5)Single cell gel electrophoresis shows that coelomocyte DNA of the earthworms was damaged in the presence of DEHP,regardless of concentration and Olive tail moment increased with the rising DEHP concentration,which suggests that DNA damage caused by DEHP is dose-dependent.【Conclusion】 Once DEHP enters earthworms,it activates the defense system of antioxidant enzymes(SOD,CAT and POD)and GST detoxification enzyme in the earthworms to protect the organisms from injury.Nevertheless,DEHP still raises ROS content in the earthworms,and excessive ROS triggers peroxidation of lipids,producing MDA.ROS and MDA act jointly damaging cell DNA of the earthworms.To sum up,DEHP may cause a certain degree of damage to organisms and DNA of the earthworms,manifesting fairly strong eco-toxicological effects.Therefore,adequate attention should be paid to DEHP disposal.
【Objective】Phthalate acid esters(PAEs),also known as phthalate esters or "plasticizer",are the reaction product of phthalic anhydride and alcohol.PAEs are widely used in the production of a huge number of products such as agricultural plastic film,toys,medical bags and hoses for transfusion,food packaging,cleansers,vinyl floor and wallpaper,and personal care products.Among them,Di(2-ethylhexyl)phthalate(DEHP)is one PAE,the highest in output and consumption and also a toxic organic compound highly hazardous to human health,disturbing endocrine systems and causing cancers,mutation,oxidative damage and genetic toxicity.As DEHP is weakly bonded to the plastic matrix by hydrogen bond and van der Waals force,it is easily detached and diffused into the air,water and soil environments.Therefore,its pollution and hazard to human health has aroused more and more attention from the people.So far studies have proven that DEHP can enter organisms via a variety of ways,causing poisonous damage to tissues and organs of the organisms.As DEHP is extensively used in agriculture and hence exists in large volumes in the soil system,it is necessary to investigate its ecological and toxicological effects,such as oxidative stress and DNA damage,on typical soil animals,in the hope that the study may serve as scientific basis for reasonable assessment of soil ecological risk of DEHP.【Method】To explore eco-toxicological effects of DEHP on soil animals,Earthworms(Eisenia foetida),used as bio-indicator,were exposed to DEHP varying in concentration(0,0.1,1,10 and 50 mg kg~(-1))in artificial soil.Earthworms were sampled after 7,14,21 and 28 days of exposure for analysis of superoxide dismutase(SOD),catalase(CAT),peroxidase(POD)and glutathione(GST),which are the indices reflecting the degree of DEHP activating oxidation of earthworms,reactive oxygen species(ROS),which is the indicator of cytotoxicity of DEHP to earthworms,and malondiadehyde(MDA)and olive tail moment,which are the indices reflecting genotoxicity of the substance to earthworms.【Result】(1)DEHP,above 0.1 mg kg~(-1) in concentration,significantly enhanced the activities of SOD,CAT,POD and GST in Eisenia foetida by a degree varying with the concentration of DEHP,which demonstrates that DEHP induces antioxidant enzymes in earthworms;(2)DEHP increased ROS concentration in earthworms as compared with control and the effect varied between treatments,displaying an apparent dose-effect relationship;(3)Comparison between the treatments shows that DEHP did have some effect on content of MDA,but the effect did not follow any specific rule and was the most obvious in earthworms exposed to DEHP,10 mg kg~(-1) and 50 mg kg~(-1) in concentration;(4)In the presence of DEHP.GST in the earthworms was apparently activated,except for the treatment of 0.1 mg kg~(-1),where the earthworms did not have much difference from those in control.The observation demonstrates that DEHP,higher than 0.1 mg kg~(-1) in concentration induces GST in the earthworms;(5)Single cell gel electrophoresis shows that coelomocyte DNA of the earthworms was damaged in the presence of DEHP,regardless of concentration and Olive tail moment increased with the rising DEHP concentration,which suggests that DNA damage caused by DEHP is dose-dependent.【Conclusion】 Once DEHP enters earthworms,it activates the defense system of antioxidant enzymes(SOD,CAT and POD)and GST detoxification enzyme in the earthworms to protect the organisms from injury.Nevertheless,DEHP still raises ROS content in the earthworms,and excessive ROS triggers peroxidation of lipids,producing MDA.ROS and MDA act jointly damaging cell DNA of the earthworms.To sum up,DEHP may cause a certain degree of damage to organisms and DNA of the earthworms,manifesting fairly strong eco-toxicological effects.Therefore,adequate attention should be paid to DEHP disposal.
引文
[1]He L,Gielen G,Bolan N S,et al.Contamination and remediation of phthalic acid esters in agricultural soils in China:Areview.Agronomy for Sustainable Development,2015,35:519—534
[2]Daiem M MA,Rivera-Utrilla J,Ocampo-Pérez R,et al.Environmental impact of phthalic acid esters and their removal from water and sediments by different technologies-A review.Journal of Environmental Management,2012,109:164—178
[3]黄慧娟,蔡全英,吕辉雄,等.土壤-蔬菜系统中邻苯二甲酸酯的研究进展.广东农业科学,2011(9):50—53Huang H J,Cai Q Y,LüH X,et al.Progresses on phthalic acid esters in the soil-vegetable system(In Chinese).Guangdong Agricultural Sciences,2011(9):50—53
[4]杨杉,吕圣红,汪军,等.酞酸酯在土壤中的环境行为与健康风险研究进展.中国生态农业学报,2016,24(6):695—703Yang S,LüS H,Wang J,et al.Environmental fate and health risks of phthalate acid esters in soils:A review(In Chinese).Chinese Journal of EcoAgriculture,2016,24(6):695—703
[5]夏庆兵.邻苯二甲酸二(2-乙基己)酯对土壤微生物群落结构多样性的影响.山东泰安:山东农业大学,2016Xia Q B.Effects of di-(2-ethylhcxyl)phthalate on the diversities of microbial community in soil(In Chinese).Tai'an,Shandong:Shandong Agricultural University,2016
[6]蔡全英,莫测辉,李云辉,等.广州、深圳地区蔬菜生产基地土壤中邻苯二甲酸酯(PAEs)研究.生态学报,2005,25(2):283—288Cai Q Y,Mo C H,Li Y H,et al.The study of PAEs in soils from typical vegetable fields in areas of Guangzhouand Shenzhen,South China(In Chinese).Acta Ecologica Sinica,2005,25(2):283—288
[7]陈永山,骆永明,章海波,等.设施菜地土壤酞酸酯污染的初步研究.土壤学报,2011,48(3):516—523Chen Y S,Luo Y M,Zhang H B,et al.Preliminary study on PAEs pollution of greenhouse soils(In Chinese).Acta Pedologica Sinica,2011,48(3):516—523
[8]Wang J,Luo Y M,Teng Y,et al.Soil contamination by phthalate esters in Chinese intensive vegetable production systems with different modes of use of plastic film.Environmental Pollution,2013,180:265—273
[9]秦华,林先贵,尹睿,等.接种降解菌对土壤中邻苯二甲酸二异辛酯降解的影响.应用与环境生物学报,2006,12(6):842—845Qin H,Lin X G,Yin R,et al.Degradation of di-(2-Ethylhexyl)phthalatein soil by inoculating microorganisms(In Chinese).Journal of Applied and Environmental Biology,2006,12(6):842—845
[10]Wang X H,Yuan X,Hou Z G,et al.Effect of di-(2-ethylhexyl)phthalate(DEHP)on microbial biomass C and enzymatic activities in soil.European Journal of Soil Biology,2009,45(4):370—376
[11]蔡文贵,秦洁芳,贾晓平,等.邻苯二甲酸二乙基己酯对翡翠贻贝(Perna viridis)生化指标的影响.生态学杂志,2012,31(1):100—105Cai W G,Qin J F,Jia X P,et al.Effects of di-(2-ethylhexyl)phthalate on the biochemical indices of green mussel(Perna viridis)(In Chinese).Chinese Journal of Ecology,2012,31(1):100—105
[12]王莉,郭靖,韩冰,等.邻苯二甲酸二乙基己酯(DEHP)对小鼠的氧化损伤及黄豆异黄酮对其保护作用的研究.生态毒理学报,2012,7(3):268—276Wang L,Guo J,Han B,et al.Oxidative damage induced by di-(2-ethylhexyl)phthalate and protective effects of soybean isoflavones in mice(In Chinese).Asian Journal of Ecotoxicology,2012,7(3):268—276
[13]颜增光,何巧力,李发生.蚯蚓生态毒理试验在土壤污染风险评价中的应用.环境科学研究,2007,20(1):134—142Yan Z G,He Q L,Li F S.The use of earthworm ecotoxicological test in risk assessment of soil contamination(In Chinese).Rresearch of Environmental Sciences,2007,20(1):134—142
[14]王广驰.烯啶虫胺和啶虫脒对蚯蚓的氧化胁迫及基因毒性.山东泰安:山东农业大学,2015Wang G C.Oxidative stress and genetoxicity induced by nitenpyram and acetamiprid exposure in earthworm(In Chinese).Tai'an,Shandong:Shandong Agricultural University,2015
[15]卢明珠,吕宪国,管强,等.蚯蚓对土壤温室气体排放的影响及机制研究进展.土壤学报,2015,52(6):1209—1225Lu M Z,LüX G,Guan Q,et al.Advancement in study on effect of earthworm on greenhouse gas emission in soil and its mechanism(In Chinese).Acta Pedologica Sinica,2015,52(6):1209—1225
[16]周垂帆.重金属和草甘膦复合污染生态毒理研究.南京:南京林业大学,2013Zhou C F.Study of the ecology toxic effects of single and combined pollution between heavy metal and glyphosate(In Chinese).Nanjing:Nanjing Forestry University,2013
[17]Liu W,Zhu L S,Wang J,et al.Assessment of the genotoxicity of endosulfan in earthworm and white clover plants using the comet assay.Archives of Environmental Contamination and Toxicology,2009,56:742—746
[18]Rastogi R P,Singh S P,H?der D P,et al.Detection of reactive oxygen species(ROS)by the oxidantsensing probe 2',7'-dichlorodihydrofluorescein diacetate in the cyanobacterium Anabaena variabilis PCC 7937.Biochemical and Biophysical Research Communications,2010,397:603—607
[19]Dikalov S I,Harrison D G.Methods for detection of mitochondrial and cellular reactive oxygen species.Antioxidants&Redox Signaling,2014,20(2):372—382
[20]Song Y,Zhu L S,Wang J,et al.DNA damage and effects on antioxidative enzymes in earthworm(Eisenia foetida)induced by atrazine.Soil Biology&Biochemistry,2009,41:905—909
[21]Song Y,Zhu L S,Xie H,et al.Effects of atrazine on DNA damage and antioxidative enzymes in Vicia faba.Environmental Toxicology and Chemistry,2009,28(5):1059—1062
[22]殷健.重金属对斑马鱼的毒性效应及作用机制研究.北京:北京协和医学院,2014Yin J.Studies on toxic effect and mechanism of heavymetals on zebrafish(In Chinese).Beijing:Peking Union Medical College,2014
[23]郑丽萍,冯艳红,赵欣,等.氯丹和灭蚁灵污染场地土壤对蚯蚓的毒性效应研究.农业环境科学学报,2010,29(10):1924—1929Deng L P,Feng Y H,Zhao X,et al.Toxicity effects of chlordane and mirex contaminated soil on earthworm(Eisenia foetida)(In Chinese).Journal of AgroEnvironment Science,2010,29(10):1924—1929
[24]Mittler R.Oxidative stress,antioxidants and stress tolerance.Trends in Plant Science,2002,7(9):405—410
[25]Wang X F,Xing M L,Shen Y,et al.Oral administration of Cr(VI)induced oxidative stress,DNA damage and apoptotic cell death in mice.Toxicology,2006,228(1):16—23
[26]陈汉,王慧君,李学锋,等.甲基苯丙胺对大鼠脑组织中NO,SOD和MDA的影响.中国药物依赖性杂志,2007,16(2):102—104Chen H,Wang H J,Li X F,et al.Change of NO,SOD and MDA in methamphetamine-treated rat brain and theircorrelation(In Chinese).Chinese Journal of Drug Dependence,2007,16(2):102—104
[27]Wang X,Jiang L J,Ge L,et al.Oxidative DNA damage induced by di-(2-ethylhexyl)phthalate in HEK-293 cell line.Environmental Toxicology and Pharmacology,2015,39(3):1099—1106
[28]赵春风,刁晓平,曹佳,等.邻苯二甲酸二(2-乙基己)酯(DEHP)对马氏珠母贝(Pinctada maetensii)胚胎发育的影响及DNA损伤.海洋环境科学,2015,34(4):546—552Zhao C F,Diao X P,Cao J,et al.Effects of DEHP on embryonic development and DNA damage of Pinctada maetensii(In Chinese).Marine Environmental Science,2015,34(4):546—552
[2]Daiem M MA,Rivera-Utrilla J,Ocampo-Pérez R,et al.Environmental impact of phthalic acid esters and their removal from water and sediments by different technologies-A review.Journal of Environmental Management,2012,109:164—178
[3]黄慧娟,蔡全英,吕辉雄,等.土壤-蔬菜系统中邻苯二甲酸酯的研究进展.广东农业科学,2011(9):50—53Huang H J,Cai Q Y,LüH X,et al.Progresses on phthalic acid esters in the soil-vegetable system(In Chinese).Guangdong Agricultural Sciences,2011(9):50—53
[4]杨杉,吕圣红,汪军,等.酞酸酯在土壤中的环境行为与健康风险研究进展.中国生态农业学报,2016,24(6):695—703Yang S,LüS H,Wang J,et al.Environmental fate and health risks of phthalate acid esters in soils:A review(In Chinese).Chinese Journal of EcoAgriculture,2016,24(6):695—703
[5]夏庆兵.邻苯二甲酸二(2-乙基己)酯对土壤微生物群落结构多样性的影响.山东泰安:山东农业大学,2016Xia Q B.Effects of di-(2-ethylhcxyl)phthalate on the diversities of microbial community in soil(In Chinese).Tai'an,Shandong:Shandong Agricultural University,2016
[6]蔡全英,莫测辉,李云辉,等.广州、深圳地区蔬菜生产基地土壤中邻苯二甲酸酯(PAEs)研究.生态学报,2005,25(2):283—288Cai Q Y,Mo C H,Li Y H,et al.The study of PAEs in soils from typical vegetable fields in areas of Guangzhouand Shenzhen,South China(In Chinese).Acta Ecologica Sinica,2005,25(2):283—288
[7]陈永山,骆永明,章海波,等.设施菜地土壤酞酸酯污染的初步研究.土壤学报,2011,48(3):516—523Chen Y S,Luo Y M,Zhang H B,et al.Preliminary study on PAEs pollution of greenhouse soils(In Chinese).Acta Pedologica Sinica,2011,48(3):516—523
[8]Wang J,Luo Y M,Teng Y,et al.Soil contamination by phthalate esters in Chinese intensive vegetable production systems with different modes of use of plastic film.Environmental Pollution,2013,180:265—273
[9]秦华,林先贵,尹睿,等.接种降解菌对土壤中邻苯二甲酸二异辛酯降解的影响.应用与环境生物学报,2006,12(6):842—845Qin H,Lin X G,Yin R,et al.Degradation of di-(2-Ethylhexyl)phthalatein soil by inoculating microorganisms(In Chinese).Journal of Applied and Environmental Biology,2006,12(6):842—845
[10]Wang X H,Yuan X,Hou Z G,et al.Effect of di-(2-ethylhexyl)phthalate(DEHP)on microbial biomass C and enzymatic activities in soil.European Journal of Soil Biology,2009,45(4):370—376
[11]蔡文贵,秦洁芳,贾晓平,等.邻苯二甲酸二乙基己酯对翡翠贻贝(Perna viridis)生化指标的影响.生态学杂志,2012,31(1):100—105Cai W G,Qin J F,Jia X P,et al.Effects of di-(2-ethylhexyl)phthalate on the biochemical indices of green mussel(Perna viridis)(In Chinese).Chinese Journal of Ecology,2012,31(1):100—105
[12]王莉,郭靖,韩冰,等.邻苯二甲酸二乙基己酯(DEHP)对小鼠的氧化损伤及黄豆异黄酮对其保护作用的研究.生态毒理学报,2012,7(3):268—276Wang L,Guo J,Han B,et al.Oxidative damage induced by di-(2-ethylhexyl)phthalate and protective effects of soybean isoflavones in mice(In Chinese).Asian Journal of Ecotoxicology,2012,7(3):268—276
[13]颜增光,何巧力,李发生.蚯蚓生态毒理试验在土壤污染风险评价中的应用.环境科学研究,2007,20(1):134—142Yan Z G,He Q L,Li F S.The use of earthworm ecotoxicological test in risk assessment of soil contamination(In Chinese).Rresearch of Environmental Sciences,2007,20(1):134—142
[14]王广驰.烯啶虫胺和啶虫脒对蚯蚓的氧化胁迫及基因毒性.山东泰安:山东农业大学,2015Wang G C.Oxidative stress and genetoxicity induced by nitenpyram and acetamiprid exposure in earthworm(In Chinese).Tai'an,Shandong:Shandong Agricultural University,2015
[15]卢明珠,吕宪国,管强,等.蚯蚓对土壤温室气体排放的影响及机制研究进展.土壤学报,2015,52(6):1209—1225Lu M Z,LüX G,Guan Q,et al.Advancement in study on effect of earthworm on greenhouse gas emission in soil and its mechanism(In Chinese).Acta Pedologica Sinica,2015,52(6):1209—1225
[16]周垂帆.重金属和草甘膦复合污染生态毒理研究.南京:南京林业大学,2013Zhou C F.Study of the ecology toxic effects of single and combined pollution between heavy metal and glyphosate(In Chinese).Nanjing:Nanjing Forestry University,2013
[17]Liu W,Zhu L S,Wang J,et al.Assessment of the genotoxicity of endosulfan in earthworm and white clover plants using the comet assay.Archives of Environmental Contamination and Toxicology,2009,56:742—746
[18]Rastogi R P,Singh S P,H?der D P,et al.Detection of reactive oxygen species(ROS)by the oxidantsensing probe 2',7'-dichlorodihydrofluorescein diacetate in the cyanobacterium Anabaena variabilis PCC 7937.Biochemical and Biophysical Research Communications,2010,397:603—607
[19]Dikalov S I,Harrison D G.Methods for detection of mitochondrial and cellular reactive oxygen species.Antioxidants&Redox Signaling,2014,20(2):372—382
[20]Song Y,Zhu L S,Wang J,et al.DNA damage and effects on antioxidative enzymes in earthworm(Eisenia foetida)induced by atrazine.Soil Biology&Biochemistry,2009,41:905—909
[21]Song Y,Zhu L S,Xie H,et al.Effects of atrazine on DNA damage and antioxidative enzymes in Vicia faba.Environmental Toxicology and Chemistry,2009,28(5):1059—1062
[22]殷健.重金属对斑马鱼的毒性效应及作用机制研究.北京:北京协和医学院,2014Yin J.Studies on toxic effect and mechanism of heavymetals on zebrafish(In Chinese).Beijing:Peking Union Medical College,2014
[23]郑丽萍,冯艳红,赵欣,等.氯丹和灭蚁灵污染场地土壤对蚯蚓的毒性效应研究.农业环境科学学报,2010,29(10):1924—1929Deng L P,Feng Y H,Zhao X,et al.Toxicity effects of chlordane and mirex contaminated soil on earthworm(Eisenia foetida)(In Chinese).Journal of AgroEnvironment Science,2010,29(10):1924—1929
[24]Mittler R.Oxidative stress,antioxidants and stress tolerance.Trends in Plant Science,2002,7(9):405—410
[25]Wang X F,Xing M L,Shen Y,et al.Oral administration of Cr(VI)induced oxidative stress,DNA damage and apoptotic cell death in mice.Toxicology,2006,228(1):16—23
[26]陈汉,王慧君,李学锋,等.甲基苯丙胺对大鼠脑组织中NO,SOD和MDA的影响.中国药物依赖性杂志,2007,16(2):102—104Chen H,Wang H J,Li X F,et al.Change of NO,SOD and MDA in methamphetamine-treated rat brain and theircorrelation(In Chinese).Chinese Journal of Drug Dependence,2007,16(2):102—104
[27]Wang X,Jiang L J,Ge L,et al.Oxidative DNA damage induced by di-(2-ethylhexyl)phthalate in HEK-293 cell line.Environmental Toxicology and Pharmacology,2015,39(3):1099—1106
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