维甲酸对早期发育斑马鱼的心脏与骨骼毒性研究

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LC50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 μmol/L). The morphology of embryos and larvae hearts were observed 24,48 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(0.1,1,10,25,50μmol/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 ob-vious 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,yolksa-cedema,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 LC50 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 con-centration of RA. CONCLUSIONS:RA can cause cardiac and skeletal toxicity in Danio rerio embryo and larvae,which is positive-ly associated with the concentration of RA.