| 五种农药及其二元、三元组合对人肝癌HepG2细胞的联合毒性 |
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| 引用本文： | 王天彩,陈晨,马朦朦,杨茜,李耘,钱永忠. 五种农药及其二元、三元组合对人肝癌HepG2细胞的联合毒性[J]. 农药学学报, 2021, 23(3): 499-508. DOI: 10.16801/j.issn.1008-7303.2021.0021 |
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| 作者姓名： | 王天彩 陈晨 马朦朦 杨茜 李耘 钱永忠 |
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| 作者单位： | 中国农业科学院农业质量标准与检测技术研究所,北京100081 |
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| 基金项目： | 国家重点研发计划 (2018YFC1603000)；国家自然科学基金项目 (31771894) |
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| 摘 要： | 为研究农产品中农药混合污染的联合毒性,选取生菜中应用广泛、广谱性强的5种农药(苯醚甲环唑、氯氰菊酯、烯酰吗啉、氯氟氰菊酯和啶虫脒)及其高频检出的二元、三元农药组合为研究对象,采用CCK-8(Cell Counting Assay Kit-8)法探究农药单剂及混合物对人肝癌HepG2细胞增殖的抑制毒性,基于单个农药剂量效...
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| 关 键 词： | 农药联合毒性 人肝癌细胞 细胞增殖 联合指数 细胞凋亡 |
| 收稿时间： | 2020-07-16 |
Combined toxicity of five pesticides and their binary and ternary mixtures on HepG2 cells |
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| Affiliation: | Institute of Quality Standards and Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing 100081, China |
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| Abstract: | To study the combined toxicity of pesticides mixtures in agricultural products, five pesticides with wide application and broad spectrum, including difenoconazole, cypermethrin, dimethomorph, cyhalothrin, acetamiprid, and relevant binary and ternary combinations with high exposure levels in lettuce were selected. Cell Counting Assay Kit-8 method was used to determine the proliferation inhibition toxicity of single pesticide and mixtures on HepG2 hepatocytes. The combined effects were assessed by three prediction models, concentration addition, independent action and combination index, based on the concentration-response curve of individual pesticides. Then the induction effects of two ternary combinations on cell apoptosis were analyzed by High Content Screening. The results showed that the effects of different pesticide combinations on the cell growth inhibitory rate of HepG2 cells were different, all of which had obvious concentration-response relationship. Among them, difenoconazole had a strong inhibition of proliferation toxicity, and the median effective concentration was 24.72 μmol/L. The order of toxicity for the other individual pesticides was ranked as dimethomorph > cyhalothrin > acetamiprid > cypermethrin. The combined toxicity of the pesticide mixtures was determined by the combination index. The results showed that with the increase of pesticide concentrations, the combined effects of all binary combinations changed from antagonism to synergy. While in ternary mixtures, including dimethomorph + difenoconazole + cypermethrin and dimethomorph + cypermethrin + acetamiprid, the combined effects changed from synergy to antagonism when the growth inhibition effect were 60% and 18%, respectively. In addition, the dose reduction index was used to quantify the synergistic effect between pesticide mixtures, indicating that cypermethrin played a greater role in the synergistic effect of the above two ternary combinations. In the measurement of apoptosis toxicity, the apoptotic rate of HepG2 cells significantly increased in the groups treated by individual pesticides in a concentration-dependent manner. The apoptotic rate induced by the ternary combination of dimethomorph + cypermethrin + acetamiprid was significantly increased in concentration between of 20.02-80.10 μmol/L. This work confirmed that the toxicities of proliferation inhibition and apoptosis induction of different pesticides mixtures on HepG2 cells both showed obvious concentration-response relationship, which could provide a scientific basis for toxicity evaluation and risk monitoring of pesticides mixtures. The combination index model as an effective ecotoxicological risk assessment tool can quantitatively describe the interactions of pesticide mixtures under different components and concentrations, which can predict the combined toxicity of pesticide mixtures more accurately compared with the concentration addition model and the independent action model. |
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