氯化镉对斑马鱼幼鱼视网膜发育的影响  

Effects of Cadmium Chloride on Retinal Development in Larval Zebrafish

作　　者：唐颖 王丹 彭星宇 曹晗霖 谭海萍 张春慧 谷红梅[1] 郭庶 Tang Ying;Wang Dan;Peng Xingyu;Cao Hanlin;Tan Haiping;Zhang Chunhui;Gu Hongmei;Guo Shu(School of Public Health,Mudanjiang Medical College,Mudanjiang 157011,China;College of Engineering,Guangzhou College of Technology and Business,Foshan 528138,China;Technical Centre for Soil,Agriculture and Rural Ecology and Environment,Ministry of Ecology and Environment,Beijing 100012,China;School of Public Health,China Medical University,Shenyang 110122,China;South China Institute of Environmental Sciences,Ministry of Ecology and Environment,National Key Laboratory of Environmental Pollution Health Risk Assessment for Environmental Protection,Guangzhou 510535,China)

机构地区：[1]牡丹江医学院公共卫生学院,牡丹江157011 [2]广州工商学院工学院,佛山528138 [3]生态环境部土壤与农业农村生态环境监管技术中心,北京100012 [4]中国医科大学公共卫生学院,沈阳110122 [5]生态环境部华南环境科学研究所,国家环境保护环境污染健康风险评价重点实验室,广州510535

出　　处：《生态毒理学报》2023年第3期418-429,共12页Asian Journal of Ecotoxicology

基　　金：国家重点研发计划课题(2019YFC1803401,2020YFC1806303)。

摘　　要：镉(cadmium,Cd)可引起斑马鱼眼部毒性效应和行为异常。目前,Cd导致斑马鱼的视网膜发育缺陷机制仍不明确。研究急性镉暴露对斑马鱼视网膜发育的影响。以斑马鱼作为模式生物,将6 hpf(hours post fertilization,hpf)的斑马鱼胚胎暴露于0(对照)、0.2、0.4、0.8、1.6 mg·L^(-1)氯化镉中,观察120 hpf斑马鱼幼鱼的生长状况,并统计胚胎存活率、孵化率和畸形率。后续将斑马鱼胚胎分别暴露于0(对照)、0.8 mg·L^(-1)氯化镉、0.8 mg·L^(-1)氯化镉与0.4 mg·L^(-1)N-乙酰半胱氨酸(NAC)混合物及0.4 mg·L^(-1)NAC,观察120 hpf斑马鱼幼鱼生长发育情况和检测氧化应激水平,镜下观察视网膜组织石蜡切片,了解视网膜结构变化;通过运动行为学实验,探讨对视神经的影响;采用qRT-PCR方法检测视网膜发育相关基因,揭示Cd对斑马鱼幼鱼视网膜的发育毒性及分子机制。斑马鱼胚胎的存活率、孵化率在0.8 mg·L^(-1)氯化镉的暴露下开始显著下降(P<0.01),畸形率显著提高(P<0.01)。0.8 mg·L^(-1)氯化镉与0.4 mg·L^(-1)NAC联合暴露后,幼鱼存活率和孵化率显著提高(P<0.05),畸形率显著下降(P<0.05)。与对照组相比,0.8 mg·L^(-1)氯化镉暴露组斑马鱼幼鱼的眼长与身长比(7.35±0.58)%显著下降(P<0.05),视网膜分层被破坏,对光敏感性降低,活动能力改变,视网膜发育基因opn1sw1、vsx1、brn3b、pax6表达水平显著下调(P<0.05)。与对照组相比,0.8 mg·L^(-1)氯化镉暴露组120 hpf斑马鱼幼鱼体内活性氧(ROS)含量和丙二醛(MDA)含量显著性升高(P<0.05),超氧化物歧化酶(SOD)活性显著下降(P<0.05);与0.8 mg·L^(-1)氯化镉暴露组相比,0.8 mg·L^(-1)氯化镉与0.4 mg·L^(-1)NAC联合暴露的谷胱甘肽S-转移酶(GST)含量显著升高(P<0.01)。Cd暴露可影响斑马鱼幼鱼及胚胎的氧化应激、神经发育等,并通过降低视网膜发育相关基因的表达水平干扰斑马鱼幼鱼视网膜发育,进一步造成斑马鱼幼鱼的视觉Cadmium(Cd)exposure may induce ocular toxicity and behavioral abnormalities in zebrafish.However,the mechanism of Cd-induced retinal developmental defects in zebrafish remains unclear.To access the impact of acute cadmium exposure on retinal development in zebrafish.Using zebrafish as a model organism,zebrafish embryos of 6 hours post fertilization(hpf)were exposed to cadmium chloride at different concentrations(0,0.2,0.4,0.8,1.6 mg·L^(-1))for 120 h.The growth status of juvenile fish,and the embryo survival rate,hatching rate and deformity rate were evaluated.Subsequently,cadmium chloride(0.8 mg·L^(-1))and N-acetylcysteine(NAC)(0.4 mg·L^(-1))were administered individually or combined to zebrafish embryos from 6 hpf until 120 hpf.Movement behavior experiment was tested.We also detected the level of growth,oxidative stress,the change of retinal structure and retinal development-related genes.The rates of survival and hatching of zebrafish embryos were significant reduced(P<0.01),while the rate of deformity was significant increased(P<0.01)in 0.8 mg·L^(-1)cadmium chloride group.Incontrast,the rates of survival and hatching of zebrafish embryos were increased and the rate of deformity was decreased in combined group.We observed a higher ratio of eye/body length,destroyed retinal delamination,lower sensitivity to light and lower activity ability of zebrafish embryos in cadmium chloride group than control.In addition,the expression levels of retinal development genes opn1sw1,vsx1,brn3b,and pax6 were significantly down-regulated(P<0.05)in zebrafish embryos after exposure to cadmium chloride.Similarly,the level of reactive oxygen species(ROS)and malondialdehyde(MDA)increased while deformity rate decreased significantly in 120 hpf zebrafish larvae after exposure to 0.8 mg·L^(-1)cadmium chloride.Compared to individual exposure to cadmium chloride,the content of glutathione S-transferase(GST)was significantly increased(P<0.01)in 120 hpf zebrafish larvae after exposure combined with NAC.Zebrafish embryos exposure to cad

关 键 词：镉(CD) 斑马鱼 视网膜 视觉行为 组织病理学 分子机制 

分 类 号：X171.5[环境科学与工程—环境科学]