基于傅里叶变换红外光谱技术分析壬基酚和双酚A对斑马鱼胚胎的影响  

Effects of Nonylphenol and Bisphenol A on Zebrafish Eleutheroembryos Based on Fourier Transform Infrared Spectroscopy

作　　者：刘芳 田斐 史文俊 Liu Fang;Tian Fei;Shi Wenjun(School of Geography,South China Normal University,Guangzhou 510631,China;South China Sea Fisheries Research Institute,Chinese Academy of Fishery Science,Guangdong Provincial Key Laboratory of Fishery Ecology and Environment,Scientific Observing and Experimental Station of South China Sea Fishery Resource and Environment,Ministry of Agriculture and Rural Affairs,Scientific Observation and Research Field Station of Pearl River Estuary Ecosystem,Guangdong Province,Guangzhou 510300,China;Guangzhou Institute of Geochemistry,Chinese Academy of Sciences,Guangzhou 510640,China;SCNU Environmental Research Institute,Guangdong Provincial Key Laboratory of Chemical Pollution and Environmental Safety&MOE Key Laboratory of Environmental Theoretical Chemistry,South China Normal University,Guangzhou 510006,China)

机构地区：[1]华南师范大学地理科学学院,广州510631 [2]中国水产科学研究院南海水产研究所,广东省渔业生态环境重点实验室,农业农村部南海渔业资源环境科学观测实验站,广东珠江口生态系统野外科学观测研究站,广州510300 [3]中国科学院广州地球化学研究所,广州510640 [4]华南师范大学环境研究院广东省化学污染与环境安全重点实验室,华南师范大学环境理论化学教育部重点实验室,广州510006

出　　处：《生态毒理学报》2022年第1期224-235,共12页Asian Journal of Ecotoxicology

基　　金：广东省自然科学基金资助项目(2017A030313220,2014A030310236);国家自然科学基金资助项目(41807480)。

摘　　要：壬基酚(4-NP)和双酚A(BPA)是2种在环境中普遍检出的雌激素物质,已经在地表水中广泛检出,且浓度很高。壬基酚和双酚A在地表水中检测浓度最高分别达到了644μg·L^(-1)和12μg·L^(-1)。目前有大量研究表明壬基酚和双酚A具有环境内分泌干扰效应,但是较少研究利用傅里叶变换红外光谱技术分析这2种物质对斑马鱼胚胎生理生化的影响。傅里叶变换红外光谱作为一种基于官能团和极性键振动结构分析技术,已广泛用于大分子化合物结构分析以及蛋白质的二级结构解析,是获取分子结构信息的有力工具,经常用于毒理学领域。本研究基于傅里叶变换红外光谱技术,分析了壬基酚和双酚A对斑马鱼胚胎的毒性。结果表明,当壬基酚≥322μg·L^(-1)时,斑马鱼胚胎72 hpf孵化率分别只有33%、5%、0%、0%和0%;同时,暴露96 h时,可显著降低斑马鱼胚胎的存活率和增加致畸指数,且呈浓度依赖特征。特别是849.5μg·L^(-1)和1038.5μg·L^(-1)壬基酚暴露后,斑马鱼胚胎全部死亡。类似地,当双酚A≥5.55 mg·L^(-1)时,斑马鱼胚胎72 hpf孵化率分别只有30%、0%、0%和0%;暴露96 h时,双酚A≥7.5 mg·L^(-1)也导致斑马鱼胚胎的存活率显著下降和增加致畸指数,并随着浓度升高毒性增大。以胚胎死亡率和致畸形指数作为效应终点,壬基酚对斑马鱼胚胎的96 h-LC_(50)和EC_(50)分别为481.7μg·L^(-1)和362.6μg·L^(-1);双酚A对斑马鱼胚胎的96 h-LC_(50)和EC_(50)为8.8 mg·L^(-1)和7.9 mg·L^(-1)。红外光谱分析显示,当壬基酚≥158.5μg·L^(-1)时,对酰胺Ⅱ、脂质、蛋白磷酸化等波数有显著影响;当双酚A≥1.75 mg·L^(-1)时,对酰胺Ⅰ、酰胺Ⅱ、脂质和碳水化合物等波数有显著影响。上述研究结果显示壬基酚和双酚A对斑马鱼存活率、畸形率以及生理生化均有显著的影响。Nonylphenol(4-NP)and bisphenol A(BPA)are two topical environment estrogen substances,and are widely detected in surface water.Furthermore,the measured concentrations of 4-NP(up to 644μg·L^(-1))and BPA(up to 12μg·L^(-1))are often high in the surface water.Early studies mainly focused on the endocrine disrupting effects of 4-NP and BPA.However,few studies investigate the effects of 4-NP and BPA on zebrafish eleutheroembryos based on Fourier Transform Infrared(FTIR)spectroscopy.FTIR spectroscopy is a sensor-based tool correlating the structure of biomolecules in different biological systems.As different chemical bonds of biochemical samples absorb light in the mid-infrared region,the application of FTIR spectroscopy help to determine the chemical structure of molecules in biochemical samples.The FTIR spectroscopy is frequently used in toxicology.In the present study,the zebrafish eleutheroembryos were exposed to 4-NP and BPA for 96 hours post-fertilization(hpf).The results showed that 72 h-hatching rates were 33%,5%,0%,0%and 0%,respectively,when the concentrations of 4-NP≥332μg·L^(-1).Meanwhile,4-NP significantly increased the mortality rates,and elevated the fish teratogenicity index at 96 hpf.Of note,all zebrafish eleutheroembryos died in 849.5μg·L^(-1) and 1038.5μg·L^(-1)4-NP treatments.Similarly,72 h-hatching rates were 30%,0%,0%and 0%,respectively,when the concentrations of BPA≥5.55 mg·L^(-1).BPA significantly increased the mortality rates,and elevated the fish teratogenicity index at 96 hpf,when the concentrations of BPA≥7.5 mg·L^(-1).Based on the mortality rate and fish teratogenicity index,the 96 h-LC_(50) and 96 h-EC_(50) of 4-NP were 481.7μg·L^(-1) and 362.6μg·L^(-1),respectively,and the 96 h-LC_(50) and 96 h-EC_(50) of BPA were 8.8 mg·L^(-1) and 7.9 mg·L^(-1),respectively.FITR analysis revealed that 4-NP were associated with biochemical alterations such as amideⅡ,lipids and protein phosphorylation,whereas BPA were associated with the amide I,amideⅡ,lipids and carbohydrate These

关 键 词：壬基酚 双酚A 斑马鱼 傅里叶红外光谱技术 

分 类 号：X171.5[环境科学与工程—环境科学]