“Cocktail”法研究辛硫磷对鲫鱼细胞色素P450酶活性的影响

Approach for assessing the effect of phoxim on CYP450 enzymes activities in crucian carp by “cocktail”

目的建立"Cocktail"探针药物法分析辛硫磷对鲫鱼细胞色素P450不同亚型酶系代谢活性的影响,为探究辛硫磷在鲫鱼体内的残留代谢途径及毒理学评价奠定基础。方法将鲫鱼随机分为4组,即辛硫磷染毒组、空白对照组、抑制剂对照组和诱导剂对照组,连续给药1周后,利用液液萃取-高效液相色谱法检测鲫鱼血浆中各探针药物浓度及药动学参数。结果辛硫磷能够显著增加咖啡因在鲫鱼体内的半衰期t_(1/2)(P<05)、血药浓度与时间曲线下的面积AUC(P<0.01)以及平均驻留时间MRT(P<0.05);能够显著降低甲苯磺丁脲在鲫鱼体内的清除率CL(P<0.05)、AUC(P<0.01)以及MRT(P<0.05),氨苯砜在鲫鱼体内的t1/2(P<0.05)、AUC(P<0.05)以及MRT(P<0.01)。而对氯唑沙宗、奥美拉唑、美托洛尔3种探针药物在鲫鱼体内的药动学参数无显著影响。结论辛硫磷能够抑制鲫鱼体内CYP1A2的活性,诱导鲫鱼体内的CYP2C9、CYP3A4的活性,对鲫鱼体内CYP2E1、CYP2D6和CYP2C19的活性不具有明显的诱导或抑制作用。

Objective To establish ＂cocktail＂ method for elucidating the effects of phoxim on the in vivo metabolism of different isoforms enzyme system of cytochrome P450 in crucian carp, in order to lay the foundation for exploring the metabolic pathways and toxicology evaluation of phoxim residues in crucian carp body. Methods The crucian carp were randomly divided into 4 groups（phoxim group, control group, inhibitors group and revulsant group）. The plasma samples obtained at different intervals were analyzed by liquid-liquid extraction-high performance liquid chromatography（LLE-HPLC） after 7 d continuous dosing. Results Phoxim could significantly increase the half-life（t_（1/2））（P〈0.05）, the area under the blood drug concentration and time（AUC）（P〈0.01）, and the average resident time（MRT）（P〈0.05） of caffeine in crucian carp, and could significantly reduce the clearance（CL）（P〈0.05）, AUC（P〈0.01） and MRT（P〈0.05） of tolbutamide in crucian carp; and significantly reduce the t1/2（P〈0.05）, the AUC（P〈0.05） and the MRT（P〈0.01） of dapsone in crucian carp. The pharmacokinetic parameters of chlorzoxazone, metoprolol, omeprazole had no significant differences between phoxim group and control group. Conclusion Phoxim could strongly inhibit the activity of CYP1A2, induce the activity of CYP2C9 and CYP3A4, and had no significant effect on the activities of CYP2E1, CYP2C19 and CYP2D6 in crucian carp.