维甲酸对早期发育斑马鱼的心脏与骨骼毒性研究
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篇名: 维甲酸对早期发育斑马鱼的心脏与骨骼毒性研究
TITLE:
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、100 μmol/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、50 μmol/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。1~100 μmol/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。100 μmol/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 、23.69 μmol/L。0.1~50 μmol/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。
ABSTRACT: 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, 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 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 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.
期刊: 2016年第27卷第19期
作者: 詹扬,王长梅,韦英杰,贾晓斌
AUTHORS: ZHAN Yang,WANG Changmei,WEI Yingjie,JIA Xiaobin
关键字: 维甲酸;心脏毒性;骨骼致畸;斑马鱼;胚胎;幼鱼
KEYWORDS: Retinoic acid; Cardiac toxicity; Skeletal deformities; Danio rerio; Embryo; Larvae
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