典型手性三唑类杀菌剂与人细胞色素CYP3A4酶相互作用的分子机制研究

项目名称： 典型手性三唑类杀菌剂与人细胞色素CYP3A4酶相互作用的分子机制研究

项目编号： No.21277122

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 环境科学、安全科学

项目作者： 庄树林

作者单位： 浙江大学

项目金额： 82万元

中文摘要： 目前我国三唑类杀菌剂使用量大且残留超标严重，已引起严重的环境健康问题。国外已发现三唑类杀菌剂的致肝癌毒性，并加强了对三唑类杀菌剂的风险管理，而国内相关研究工作极少，尤其缺乏针对毒性作用分子机制的研究，这预示了开展相关研究工作的必要性和紧迫性。本项目将采用QM/MM计算方法并结合荧光寿命、细胞增殖、蛋白质印迹等实验研究8种典型手性三唑类杀菌剂与人CYP3A4酶的动态相互作用。在对映体水平上考察对映体的立体选择性差异;在分子水平上考察对人CYP3A4酶构象变化、酶活性的影响；在细胞水平上考察对HepG2细胞内CYP3A4酶表达的影响和促人肝癌HepG2细胞体外增殖效应；以期诠释手性三唑类杀菌剂毒性作用的分子机制，为环境安全和人体健康风险评估提供重要的科学数据支持。

中文关键词： 三唑杀菌剂；CYP3A4；相互作用；代谢；手性

英文摘要： The triazole fungicides were used quantitatively in China and their residues much exceeded the maximum residue limit,causing serious environmental health issues.The research abroad found the hepatocarcinogens properties of triazole fungicides and their risk management was further strengthened.However,the domestic research on toxic identification and evaluation of triazole fungicides is rare,especially there lacks the research on molecular mechanisms of action,showing the necessity and urgency to carry out relevant research.In this project,the QM/MM method together with fluorescence lifetime,cell proliferation assay and Western Blot analysis will be performed to investigate the interactions of eight typical triazole fungicides with human CYP3A4 enzyme.The focus of this project is to probe the stereoselective difference of enantiomers,to monitor conformational changes and activity change of human CYP3A4 enzyme at an atomic level of detail and to investigate the effect on CYP3A4 expression within HepG2 cell and on HepG2 cell proliferation.Our aim is to elucidate mechanism of toxic action of triazole fungicides,thus providing scientific data for risk assessment of environmental and human health hazards.

英文关键词： Triazole fungicide；CYP3A4；Interaction；Metabolism；Chirality