炔诺酮暴露下斑马鱼候选生物标志物筛选研究  

Study on the screening of biomarkers in zebrafish under Norethindrone exposure

作　　者：谭芷晴 桂林[1] 李新昌[2] 王小兰[1] TAN Zhiqing;GUI Lin;LI Xinchang;WANG Xiaolan(School of Life Science,Guangzhou University,Guangzhou 510006,China;School of Life Science,Zhaoqing University,Zhaoqing 526061,China)

机构地区：[1]广州大学生命科学学院,广州510006 [2]肇庆学院生命科学学院,肇庆526061

出　　处：《生态科学》2023年第2期35-42,共8页Ecological Science

基　　金：国家自然科学基金(41977207);广东省自然科学基金项目(2017A030313141);广州大学2019年示范性实验建设项目(SJ201904)。

摘　　要：NET(炔诺酮,Norethindrone)是一种人工合成的孕激素,被广泛应用于医学和畜牧养殖。近年来国内外不断报道的从环境中检测到的NET对鱼类的肝脏和内分泌均产生干扰效应。为了寻找水环境中的NET对鱼类影响的生物标志物(Biomarkers),以成年雌性斑马鱼为动物模型,暴露于500 ng·L^(-1)NET后60 d,采集肝脏,运用非靶向HPLC-TOF-MS的方法,分析了斑马鱼肝脏代谢物在负离子模式条件下的变化。结果表明:(1)经500 ng·L^(-1)NET暴露的处理组与0 ng·L^(-1)的对照组在PCA分析中分别聚集并明显分开,说明NET对斑马鱼的肝脏代谢存在明显干扰作用。(2)代谢通路分析说明NET暴露显著影响了斑马鱼肝脏的硫胺素代谢、甘油磷脂代谢和花生四烯酸代谢过程;另外,差异代谢物网络关系图也显示磷脂和脂肪酸成分影响较大,这与代谢物磷脂成分(PA、PC和PI等)和溶血磷脂(LysoPC)在NET处理后发生显著性变化的结果相一致,推测NET可能影响了生物膜系统的结构和功能。(3)对鉴定的差异代谢物以VIP>1作为潜在标志物的筛选依据,发现有代谢物3-甲基硫代丙酸和LysoPC(14:1(9Z))可能是NET对斑马鱼肝脏影响的潜在生物标志物。本研究为评价NET的生态风险提供了数据支持。NET(Norethindrone),a synthetic progesterone,is widely used in medicine and livestock farming.In recent years,it has been reported that NET has interfered with the liver and endocrine of fish.In order to explore the potential biomarkers of NET on fish in the environment,adult female zebrafish was taken as the animal model,and liver was collected from 60 days 500 ng·L^(-1) NET exposure.The changes of liver metabolites in zebrafish under negative ion mode were analyzed by non-targeted HPLC-TOF-MS method.The results showed that(1)the treatment group exposed to 500 ng·L^(-1) NET and the control group exposed to 0 ng·L^(-1) were aggregated and significantly separated in PCA analysis,indicating significant differences in liver metabolites between the two groups.(2)Metabolic pathway analysis showed that NET exposure significantly affected the metabolism of thiamine,glycerophospholipid and arachidonic acid in the zebrafish liver.The differential metabolite network diagram showed that the composition of phospholipids and fatty acids had great influence on the structure and function of the biofilm system.(3)Screening and identification of significant changes in phospholipid components(PA,PC,PI,etc.)and LysoPC in differential metabolites;VIP>1 as the basis for screening potential biomarkers,3-methylthiopropionic acid and LysoPC(14:1(9Z))were found to be potential biomarkers for the effect of NET on zebrafish liver.This research provides data support for evaluating the ecological risk of NET.

关 键 词：炔诺酮 斑马鱼 代谢组学 生物标志物 

分 类 号：X174[环境科学与工程—环境科学]