几种农药多元组合对HepG2人肝癌细胞的联合毒性效应
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摘要
多种农药残留经膳食暴露途径进入人体后,其联合毒性的潜在风险通常超过单一农药。为探明农药多残留的联合暴露毒性,采用经典MTT (噻唑蓝)比色法,测定了阿维菌素、啶虫脒、多菌灵、百菌清、毒死蜱、高效氯氟氰菊酯、咪酰胺和三唑磷8种农药多种组合方式联合暴露下对HepG2人肝癌细胞增殖的抑制活性。结果表明:联合暴露24 h后,不同种类农药组合对HepG2细胞存活率的影响存在较大差异,随着组合农药质量浓度升高,部分组合对HepG2细胞增殖的抑制效应分别呈现S型变化和线性变化。例如,二元组合(多菌灵+高效氯氟氰菊酯)在0～10.00 mg/L、三元组合(多菌灵+阿维菌素+高效氯氟氰菊酯)在0～24.12 mg/L、四元组合(三唑磷+高效氯氟氰菊酯+多菌灵+阿维菌素)在0～47.46 mg/L以及五元组合(毒死蜱+阿维菌素+啶虫咪+高效氯氟氰菊酯+三唑磷)在0～62.80 mg/L暴露条件下其抑制效应表现为S型变化,即随着组合农药质量浓度增大,细胞存活率先迅速降低,之后则缓慢降低;二元组合(多菌灵+百菌清)在0～8.46 mg/L、三元组合(毒死蜱+啶虫脒+多菌灵)在0～39.97 mg/L、四元组合(多菌灵+高效氯氟氰菊酯+百菌清+阿维菌素)在0～25.92 mg/L以及六元组合(啶虫脒+毒死蜱+三唑磷+多菌灵+高效氯氟氰菊酯+阿维菌素)在0～57.48 mg/L暴露条件下其抑制效应表现为线性变化,即随着组合农药质量浓度升高,细胞存活率呈线性降低。本研究对农产品中农药多残留的体外细胞毒性进行了评估,明确了不同农药多残留组合对HepG2人肝癌细胞增殖的抑制作用剂量-效应关系,可为进一步探索农药多残留的细胞毒性机制和开展食品安全风险评估提供依据。
After dietary exposure, residues of multiple pesticides will enter human body and cause the potential health risk, which is always worse than that caused by single pesticide.The proliferation inhibition toxicity of abamectin, acetamiprid, carbendazim, chlorothalonil, chlorpyrifos, cyhalothrin,prochloraz and triazophos on HepG2 hepatocytes was determined by the classical MTT colorimetry.The result showed that the effects of different pesticide combinations on the survival rate of HepG2 cells were different after 24 h exposure. The combined toxicity of pesticide mixtures(comprised of two components, three components, four components, five components and six components) demonstrated two types of concentration-response curves towards proliferation inhibition HepG2 cells with the increase of pesticide concentrations. One type was a S type trend, with the increase of the concentration of pesticides, the cell survival rate first rapidly decreased, then slowly decreased. The followings are examples of this type: binary combination(carbendazim + cyhalothrin) at concentration of 0-10.00 mg/L,ternary combination(carbendazim + abamectin + cyhalothrin) at concentration of 0-24.12 mg/L,quaternary combination(triazophos + cyhalothrin + carbendazim + abamectin) at concentration of 0-47.46 mg/L and five-component(chlorpyrifos + abamectin + acetamiprid + cyhalothrin + triazophos) at concentration of 0-62.80 mg/L. The other type was a linear trend, cell survival linearly decreased with the increase of the concentration of pesticides. The followings are examples of this type: binary combination(carbendazim + chlorothalonil) at concentration of 0-8.46 mg/L, ternary combination(chlorpyrifos + acetamiprid + carbendazim) at concentration of 0-39.97 mg/L, quaternary combined(carbendazim + cyhalothrin + chlorothalonil + abamectin) at concentration of 0-25.92 mg/L, and sixcomponent(acetamiprid + chlorpyrifos + triazophos + carbendazim + cyhalothrin + abamectin) at concentration of 0-57.48 mg/L. In this study, different combinations of pesticides showed different types of inhibition to the proliferation of human liver cancer cells, which provided a basis for the mechanism research and cumulative risk assessment of pesticide residues in the future.
After dietary exposure, residues of multiple pesticides will enter human body and cause the potential health risk, which is always worse than that caused by single pesticide.The proliferation inhibition toxicity of abamectin, acetamiprid, carbendazim, chlorothalonil, chlorpyrifos, cyhalothrin,prochloraz and triazophos on HepG2 hepatocytes was determined by the classical MTT colorimetry.The result showed that the effects of different pesticide combinations on the survival rate of HepG2 cells were different after 24 h exposure. The combined toxicity of pesticide mixtures(comprised of two components, three components, four components, five components and six components) demonstrated two types of concentration-response curves towards proliferation inhibition HepG2 cells with the increase of pesticide concentrations. One type was a S type trend, with the increase of the concentration of pesticides, the cell survival rate first rapidly decreased, then slowly decreased. The followings are examples of this type: binary combination(carbendazim + cyhalothrin) at concentration of 0-10.00 mg/L,ternary combination(carbendazim + abamectin + cyhalothrin) at concentration of 0-24.12 mg/L,quaternary combination(triazophos + cyhalothrin + carbendazim + abamectin) at concentration of 0-47.46 mg/L and five-component(chlorpyrifos + abamectin + acetamiprid + cyhalothrin + triazophos) at concentration of 0-62.80 mg/L. The other type was a linear trend, cell survival linearly decreased with the increase of the concentration of pesticides. The followings are examples of this type: binary combination(carbendazim + chlorothalonil) at concentration of 0-8.46 mg/L, ternary combination(chlorpyrifos + acetamiprid + carbendazim) at concentration of 0-39.97 mg/L, quaternary combined(carbendazim + cyhalothrin + chlorothalonil + abamectin) at concentration of 0-25.92 mg/L, and sixcomponent(acetamiprid + chlorpyrifos + triazophos + carbendazim + cyhalothrin + abamectin) at concentration of 0-57.48 mg/L. In this study, different combinations of pesticides showed different types of inhibition to the proliferation of human liver cancer cells, which provided a basis for the mechanism research and cumulative risk assessment of pesticide residues in the future.
引文
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[20]安丽,鲍清,施万英,等.乙羧氟草醚农药对大鼠肝脏毒性的研究[J].中国工业医学杂志,2004,17(3):181-182.AN L,BAO Q,SHI W Y,et al.Hepatoxicity of fluoroglycoften-ethyl pesticide in rats[J].Chin J Ind Med,2004,17(3):181-182.
[2]European Food Safety Authority(EFSA).The 2014 European Union report on pesticide residues in food[J].EFSA J,2016,14(10):e04611.
[3]PRUTNER W,NICKEN P,HAUNHORST E,et al.Effects of single pesticides and binary pesticide mixtures on estrone production in H295R cells[J].Arch Toxicol,2013,87(12):2201-2214.
[4]吴正军.浅谈现代农业中农药的混用[J].现代农业科技,2007(6):68-69.WU Z J.Discussion on the mix use of pesticides in modern agriculture[J].Mod Agric Sci Technol,2007(6):68-69.
[5]宋宁慧,卜元卿,单正军.农药对地表水污染状况研究概述[J].生态与农村环境学报,2010,26(增刊1):49-57.SONG N H,BU Y Q,SHAN Z J.A review of studies on pesticide pollution in surface water[J].J Ecol Rural Environ,2010,26(Suppl.l):49-57.
[6]CREMONESE C,PICCOLI C,PASQUALOTTO F,et al.Occupational exposure to pesticides,reproductive hormone levels and sperm quality in Young Brazilian men[J],Reprod Toxicol,20 17,67:174-185.
[7]孙金秀,陈波,姚佩佩.农药混剂联合毒性评价[J].卫生研究,2000,29(2):65-68.SUN J X,CHEN B,YAO P P.Assessment on acute toxicity of combined pesticides[J].J Hyg Res,2000,29(2):65-68.
[8]TAKAKURA N,SANDERS P,FES SARD V,et al.In vitro combinedcytotoxic effects of pesticide cocktails simultaneously found in the French diet[J].Food Chem Toxicol,2013,52:153-162.
[9]ILBOUDO S,FOUCHE E,RIZZATI V,et al.In vitro impact of five pesticides alone or in combination on human intestinal cell line Caco-2[J].Toxicol Rep,2014,1:474-489.
[10]SALEEM U,EJAZ S,ASHRAF M,et al.Mutagenic and cytotoxic potential of endosulfan and lambda-cyhalothrin:in vitro study describing individual and combined effects of pesticides[J].J Environ Sci,2014,26(7):1471-1479.
[11]MA M M,CHEN C,YANG G L,et al.Combined cytotoxic effects of pesticide mixtures present in the Chinese diet on human hepatocarcinoma cell line[J].Chemosphere,2016,159:256-266.
[12]FAUST M,ALTENBURGER R,BACKHAUS T,et al.Predicting the joint algal toxicity of multi-component s-triazine mixtures at loweffect concentrations of individual toxicants[J].Aquat Toxicol,2001,56(1):13-32.
[13]佟鹤芳.常见农药污染物在人体血液样本中的检测及人体负荷水平研究[D].北京:北京协和医学院,2013.TONG H F.Detection of common pesticide pollutants in human blood samples and study on human load level[D].Beijing,China:Peking Union Medical College,2013.
[14]DE P,CHATTERJEE R.Nucleolar localization of succinic dehydrogenase in human malignant cells with MTT[J].Experientia,1962,18(12):562.
[15]MOSMANN T.Rapid colorimetric assay for cellular growth and survival:application to proliferation and cytotoxicity assays[J].J Immunol Methods,1983,65(1-2):55-63.
[16]杨桂玲.农产品中农药多残留联合暴露风险评估方法研究[D].北京:中国农业科学院,2017.YANG G L.Risk assessment of combined exposure to multiple pesticide residues in agricultural products[D].Beijing,China:Chinese Academy of Agricultural Sciences,2017.
[17]RIZZATI V,BRIAND O,GUILLOU H,et al.Effects of pesticide mixtures in human and animal models:an update of the recent literature[J].Chem Interactions,2016,254:23 1-246.
[18]GORE A C, CHAPPELL V A,FENTON S E,et al.Executive summary to EDC-2:the endocrine society's second scientific statement on endocrine-disrupting chemicals[J].Endocr Rev,2015,36(6):593-602.
[19]VANDENBERG L N,COLBORN T,HAYES T B,et al.Hormones and endocrine-disrupting chemicals:low-dose effects and nonmonotonic dose responses[J].Endocr Rev,2012,33(3):378-455.
[20]安丽,鲍清,施万英,等.乙羧氟草醚农药对大鼠肝脏毒性的研究[J].中国工业医学杂志,2004,17(3):181-182.AN L,BAO Q,SHI W Y,et al.Hepatoxicity of fluoroglycoften-ethyl pesticide in rats[J].Chin J Ind Med,2004,17(3):181-182.
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