摘要
目的:研究维甲酸对早期发育斑马鱼心脏及骨骼的毒性。方法:将受精后24 h的斑马鱼胚胎作为毒性模型,分别暴露于含维甲酸0.1、1、10、25、100μmol/L的培养基中,分别于培养24、48 h后观察胚胎和幼鱼心脏形态的变化,并计算半数致死浓度(LC50);将受精后4 d的斑马鱼幼鱼作为骨骼致畸模型,分别暴露于含维甲酸0.1、1、10、25、50μmol/L的培养基中,5 d后处死,采用茜素红对各组斑马鱼骨骼染色,并以显微镜采集图像,观察骨骼染色区域差异。结果:维甲酸对斑马鱼胚胎的孵化能力造成了明显不利影响,且胚胎在维甲酸暴露过程中均产生了明显的心脏致畸现象。1~100μmol/L维甲酸均对胚胎和幼鱼产生心脏致畸效应,胚胎和幼鱼表现出的主要畸形特征有心脏形态线性化、心包水肿、卵黄囊肿胀、血细胞在心区堆积。100μmol/L维甲酸可抑制胚胎的孵化能力,且对胚胎和幼鱼造成致死效应,暴露24、48 h后LC_(50)分别为36.44、23.69μmol/L。0.1~50μmol/L维甲酸均使斑马鱼幼鱼脊柱提前骨化,且骨化程度与维甲酸浓度呈正相关。结论:维甲酸对斑马鱼胚胎和幼鱼具有心脏和骨骼毒性,且与维甲酸的浓度呈正相关。
OBJECTIVE:To study the cardiac and skeletal toxicity of retinoic acid(RA) in Danio rerio at early life stage.METHODS:Danio rerio embryos of 24 hours post fertilization(hpf)were used as toxicity model and were exposed under medium with various concentrations of RA(0.1,1,10,25,100 μ mol/L).The morphology of embryos and larvae hearts were observed24,48 h after exposed.LC50 was calculated.Danio rerio larvae of 4 days post fertilization(dpf)were used as skeletal deformity model and were exposed with a series of RA at various concentration(0.1,1,10,25,50 μmol/L).They were sacrificed 5 d later,and then Danio rerio skeleton were fixed for staining with alizarin red.The microscopic was used to observe the difference of stained skeleton area.RESULTS:RA caused significant adverse effects on hatching capabilities of Danio rerio embryos,and the obvious malformation features were produced during the culture process.1-100 μmol/L RA could cause heart malformation in Danio rerio embryos and larvae,and the main heart malformation characteristics included heart linearization,pericardial edema,yolksacedema,hemocytes accumulation incardiac region.100 μmol/L RA could inhibit the hatching capabilities of Danio rerio embryos,and caused lethal effects on embryos and larvae.The LC_(50) were 36.44,23.69 μmol/L after exposed for 24,48 h.0.1-50 μmol/L RA induced vertebral column sclerotization of Danio rerio embryos and larvae in advance,which was positively associated with the concentration of RA.CONCLUSIONS:RA can cause cardiac and skeletal toxicity in Danio rerio embryo and larvae,which is positively associated with the concentration of RA.
出处
《中国药房》
CAS
北大核心
2016年第19期2612-2614,共3页
China Pharmacy
基金
国家自然科学基金资助项目(No.81573833)
江苏省自然科学基金资助项目(No.BK20141507
BK2011866)
江苏省第十批"六大人才高峰"高层次人才选拔培养项目(No.2013-YY006)
江苏省第四期"333工程"科研资助项目(No.BRA2014348)
关键词
维甲酸
心脏毒性
骨骼致畸
斑马鱼
胚胎
幼鱼
Retinoic acid
Cardiac toxicity
Skeletal deformities
Danio rerio
Embryo
Larvae