1-萘酚胚胎期暴露对斑马鱼发育的毒性作用  

Developmental toxicity of embryonic 1-naphthol exposure in zebrafish

作　　者：任平平 舒莉萍[2] 王一慧 莫晶 周艳华[2] 何志旭[2,3] REN Pingping;SHU Liping;WANG Yihui;MO Jing;ZHOU Yanhua;HE Zhixu(Department of Pediatrics,Guizhou Medical University,Guiyang,Guizhou Province,550004;Tissue Engineering and Stem Cell Center,Guizhou Medical University,Key Laboratory of Adult Stem CellTransformation Research of Chinese Academy of Medical Sciences,Guiyang,Guizhou Province,550004;Department of Pediatrics,Zunyi Medical University,Zunyi,Guizhou Province,563006,China)

机构地区：[1]贵州医科大学儿科学教研室,贵阳550004 [2]贵州医科大学组织工程与干细胞中心,中国医学科学院成体干细胞转化研究重点实验室,贵阳550004 [3]遵义医科大学儿科学教研室,贵州遵义563006

出　　处：《第三军医大学学报》2019年第14期1321-1327,共7页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81871313);贵州省重大基础研究项目(黔科合J重大字[2015]2003);贵州省重大专项资金(黔科合重大专项字[2014]6008)~~

摘　　要：目的探讨1-萘酚(1-NAP)胚胎期暴露对斑马鱼发育的毒性作用,并对其可能的机制进行研究。方法挑选斑马鱼受精卵,在受精后24 h(hours post fertilization,hpf)进行不同浓度(0、40、80、160、200μmol/L)1-NAP暴露处理,观察并记录其处理至斑马鱼120 hpf时的畸形数和死亡数,计算畸形率、孵化率及半数致死浓度(IC50)。设置空白对照组,N-乙酰半胱氨酸(NAC)处理组,1-NAP处理组,NAC预处理组,检测幼鱼体内活性氧(ROS)、丙二醛(MDA)、还原型谷胱甘肽(GSH)和总巯基(-SH)含量以及超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性;通过Western blot法检测胚胎凋亡蛋白Caspase3、Bax的表达情况。结果计算出1-NAP在斑马鱼胚胎的IC50为111.31μmol/L,斑马鱼胚胎期暴露于高浓度1-萘酚(>80μmol/L)导致死亡率和畸形率明显增加,孵化率明显降低,表现出发育毒性;50μmol/L浓度1-NAP处理组ROS、MDA含量明显升高,GSH、-SH水平明显降低,抗氧化酶CAT、SOD的活性受到抑制(P<0.001);胚胎凋亡蛋白Caspase3、Bax表达量增加;NAC(0.5 mmol/L)预处理组斑马鱼胚胎的畸形率明显降低,ROS、MDA含量明显减少,GSH、-SH含量以及SOD、CAT活性有所恢复,凋亡蛋白Caspase3、Bax表达量明显降低。结论 1-萘酚胚胎期暴露对斑马鱼发育的毒性作用可能与1-萘酚诱导的氧化应激相关。Objective To investigate the toxic effects of embryonic exposure to 1-naphthol(1-NAP) on zebrafish development and explore the possible mechanism. Methods Fertilized zebrafish eggs were exposed to 1-NAP at the concentrations of 40, 80, 160 and 200 μmol/L on 24 h post fertilization(hpf). The numbers of zebrafish malformations and deaths were recorded at 120 hpf, and the deformity rate, hatching rate and the median lethal concentration(IC50) of 1-NAP were calculated. The hatched zebrafish with or without embryonic exposure to 50 μmol/L 1-NAP and pretreatment with N-acetylcysteine(NAC) were examined for levels of reactive oxygen species(ROS), malondialdehyde(MDA), reduced glutathione(GSH), total sulfhydryl, and activities of superoxide dismutase(SOD) and catalase(CAT);Western blotting was performed to detect the expression of Caspase-3 and Bax. Results The IC50 of 1-NAP in zebrafish embryos was 111.31 μmol/L, and the zebrafish embryos exposed to high-concentration(>80 μmol/L) 1-NAP exhibited obviously increased mortality and deformity rates with a significantly reduced hatching rate, demonstrating significant developmental toxicity of 1-NAP. Embryonic exposure to 50 μmol/L 1-NAP significantly increased the levels of ROS and MDA, lowered the levels of GSH and total sulfhydryl, suppressed the activities of antioxidant enzymes CAT and SOD(P<0.001), and decreased the expression levels of Caspase-3 and Bax in the hatched zebrafish. The zebrafish embryos pretreated with 0.5 mmol/L NAC prior to 1-NAP exposure showed a significantly decreased abnormality rate with obviously lowered ROS and MDA levels, restoration of GSH and total sulfhydryl content and SOD and CAT activities, and obviously reduced expression levels of Caspase-3 and Bax. Conclusion The developmental toxicity of 1-NAP in zebrafish embryos is probably associated with enhanced oxidative stress induced by 1-NAP.

关 键 词：斑马鱼 胚胎期 1-萘酚 发育毒性 氧化应激 

分 类 号：R321-33[医药卫生—人体解剖和组织胚胎学] R994.6[医药卫生—基础医学]