毒死蜱和克百威体外联合染毒的细胞毒性机制

发布时间：2018-09-02 09:04

【摘要】：目的研究毒死蜱(chlorpyrifos,CPF)与克百威(carbofuran,CF)单独及联合染毒对体外培养细胞株的毒性,并阐明联合作用模式。方法应用不同浓度的CPF(0、50、100、200和400μmol/L)和CF(0、25、50、100和200μmol/L)分别单独染毒大鼠嗜铬细胞瘤株PC12细胞12 h,联合效应研究分别在低剂量(CPF 50μmol/L,CF 25μmol/L)和高剂量(CPF 200μmol/L,CF 100μmol/L)水平下混合染毒。染毒结束后,采用丁酰硫代胆碱法检测乙酰胆碱酯酶(ACh E)活性,二氯荧光素二乙酸酯(DCFH-DA)荧光探针法测定细胞内活性氧(ROS)的生成,硫代巴比妥酸(TBA)法测定细胞中脂质过氧化产物丙二醛(MDA)含量,黄嘌呤氧化酶法测定细胞中超氧化物歧化酶(SOD)活性,5,5'-二硫代双硝基苯甲酸(DTNB)显色法测定细胞中谷胱甘肽过氧化物酶(GPx)活性。结果毒死蜱、克百威单独染毒时,对ACh E活性的抑制作用、PC12细胞内ROS的生成量、SOD及GPx的活性均显著高于对照组(P0.01),MDA含量无明显变化。2×2析因设计分析联合效应结果表明,较低剂量水平下,CPF与CF之间无交互作用,较高剂量水平,CPF、CF之间存在交互作用(P0.01),作用模式主要为协同作用。结论毒死蜱、克百威单独及联合染毒都具有细胞毒性,联合作用模式主要为协同作用,氧化应激可能是其产生细胞毒性的机制之一。
[Abstract]:Objective to study the toxicity of chlorpyrifos (chlorpyrifos,CPF) and carbofuran (carbofuran,CF) alone or in combination to cultured cell lines in vitro and to elucidate the combined effects. Methods Rat pheochromocytoma cell line PC12 cells were exposed to different concentrations of CPF (050 100 200 渭 mol/L and 400 渭 mol/L) and CF (0 25 渭 mol/L,CF 50 100 and 200 渭 mol/L) for 12 h, respectively. The combined effects were studied at low (CPF 50 渭 mol/L,CF 25 渭 mol/L) and high dose (CPF 200 渭 mol/L,CF 100 渭 mol/L) levels. The activity of acetylcholinesterase (ACh E) and the production of reactive oxygen species (ROS) in cells were detected by butyrylthiocholine method and dichlorofluorescein diacetate (DCFH-DA) fluorescence probe method. The content of malondialdehyde (MDA), a product of lipid peroxidation, was determined by thiobarbituric acid (TBA) assay. The activity of superoxide dismutase (SOD) in cells was determined by xanthine oxidase method. The activity of glutathione peroxidase (GPx) was determined by (DTNB) colorimetric assay. Results the inhibitory effects of chlorpyrifos and carbofuran on the activity of ACh E in PC12 cells were significantly higher than those in the control group (P0.01). The combined effect of 2 脳 2 factorial design analysis showed that the activity of ROS and GPx in PC12 cells were significantly higher than those in the control group (P0.01). There was no interaction between CF and CF at lower dose level (P0.01). Conclusion Chlorpyrifos and carbofuran alone or in combination have cytotoxicity. The mode of combined action is mainly synergistic. Oxidative stress may be one of the mechanisms of cytotoxicity of chlorpyrifos.
【作者单位】：四川大学华西公共卫生学院营养食品卫生与毒理学系;上海市食品药品检验所药理毒理室;军事医学科学院疾病预防控制所毒理学评价研究中心;
【基金】：国家“十一五”科技支撑计划(No.2006BAK02A00)
【分类号】：R99

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