辛硫磷与鲫鱼肝CYP450作用机理研究及作用位点分析

项目名称： 辛硫磷与鲫鱼肝CYP450作用机理研究及作用位点分析

项目编号： No.31471655

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 刘晓宇

作者单位： 华中农业大学

项目金额： 88万元

中文摘要： 淡水鱼类是深受我国人民喜爱的食品，其食用安全问题意义重大。而随着化学农药使用量的增加，其对淡水鱼类的安全影响日益引起关注。针对相应农药标准缺乏从而导致水产食品安全隐患的现状，本项目以有机磷农药辛硫磷为化学农药代表，以分布广泛的鲫鱼为淡水鱼类代表，开展系统的基础研究。包括农药代谢关键酶CYP450酶系的亚型鉴定，辛硫磷对CYP450亚型mRNA及蛋白表达的影响，并运用MALDI-TOF MS技术鉴定作用位点；运用AFM、CD等现代仪器技术研究辛硫磷对CYP450结构变化影响，阐释辛硫磷与CYP450作用机理。采用计算机模拟技术，构建辛硫磷与CYP450作用的3D-QSAR模型，推断辛硫磷作用位点及与CYP450酶之间的构效关系，预测辛硫磷代谢产物特征，揭示其作用途径和机理。为合理制定农药安全性标准，控制农药残留危害从而保障淡水产品食用安全奠定基础。

中文关键词： 辛硫磷；鲫鱼；CYP450；作用机理；分析

英文摘要： Freshwater fish is widely favoured by Chinese people, its edible safety is an issue with highly significance. As the increasing of the amount of chemical pesticides utilized in freshwater fish feeding, their effect on the edible safety of freshwater fish attracted more and more attention nowadays. The lack of pesticides standards have led to intensive safety risks for aquatic products. This project employed the widespread carassius auratus gibelio as the symbol specie and phoxim as the target analyte, a series of fundamental research using carassius auratus gibelio, which was polluted by phoxim, was performed. This project involved the identification of CYP450 isoforms, evaluation of effect of phoxim on mRNA expression of CYP isoforms, the interactional sites were also studied by MALDI-TOF MS technique. The affect of phoxim on structural change of protein molecular was researched by modern analysis technology including AFM, CD, to explain the mechanism of interaction between phoxim and CYP450. The 3D-QSAR model of CYP450 acting on phoxim was established by means of computer simulate technology, which was aiming to study the structure-activity relationship between phoxim and CYP450, to predict the characteristic of the major metabolite,and to indicate its pathway and mechanism. The research results could offer the theoretical basis for establishing pesticide safety standard,controlling the harm of pesticide residues to ensure the edible safety of aquatic products.

英文关键词： Phoxim;carassius-auratus-gibelio;CYP450;interactional mechanism;analysis