建鲤精密肝切片损伤模型的构建及甘草提取物抗TCDD诱导肝损伤的分子机制研究

二嗯英不仅给水产和畜牧业带来巨大的经济损失，对生态平衡和人类健康也构成严重威胁。本研究以建鲤作为研究对象，对TCDD的毒性机制及防治进行基础科学研究。体外实验用TCDD建立精密肝组织切片损伤模型，不同浓度甘草提取物处理肝切片，收集上清液测定SOD、MDA、IgM等指标；ELISA方法检测IL-1β、TNF-α等指标的水平；RT-PCR法检测精密肝组织切片中AHR、ARNT、CYP1A mRNA的表达水平；Western Blot法检测细胞色素酶CYP1A的蛋白表达水平。体内实验，将不同浓度甘草提取物添加到饲料当中，分离血清、采集肝脏并测定上述指标，同时也制备病理组织切片观察TCDD对肝脏的损伤程度，阐明甘草提取物抗TCDD诱导肝损伤及保肝作用分子机制。本项目旨在更好地研究二嗯英对鱼主要靶器官和靶基因的毒害，理解二嗯英对鱼毒性效应的机制，为控制二嗯英以及研究人、动物与环境污染的关系提供参考

TCDDD not only brought huge economic losses to the aquatic and animal husbandry, but also posed a serious threat to the ecological equilibrium and human health . In this study, Jian carp was slected for the basic scientific research of TCDD toxicity mechanism and prevention.In vitro experiments,the injury model of precision-cut liver slices were established by TCDD and treated with different concentrations of licorice extract, the culture supernatants were collected for the determination of SOD, MDA,IgM and other indicators;the mRNA expression of AHR,ARNT,CYP1A in precision-cut liver slices were determined by RT-PCR;the protein expression of cytochrome P4501A was determined by Western Blot.In vivo experiments, different concentrations of licorice extract were added to the feed, then the serum and liver were collected for the determination of above indicators. At the same time preparing pathological tissue sections, to observe the extent of liver injury by TCDD,to clarify the hepatoprotective effect molecular mechanism of licorice extract anti-TCDD-induced liver damage.This project aims to better study the poison of TCDD to the main target organs and target genes in fish, understanding the toxicity mechanism, providing a reference for controlling TCDD and studying the relationship of human, animal and environmental pollution.