孙晨　韩利文　何秋霞

[摘要] 目的 以斑马鱼作为实验动物，研究阿司咪唑的心脏毒性及其分子机制。 方法 选择受精后24 h（24 hours post fertilization）的AB系斑马鱼作为实验动物，分别暴露于不同浓度的阿司咪唑培养液中，处理24 h后，统计其死亡率并在显微镜下观察心脏形态的改变，记录心率变化，利用RT-qPCR技术检测心脏功能相关基因表达的改变。结果 随着阿司咪唑浓度的升高，斑马鱼胚胎逐渐出现死亡、心囊水肿、心脏畸形、心率下降等情况；7种心脏功能相关基因（CAV1.3a、CAV1.3b、HCN2、HCN4、KCNH2a、KCNQ1和KCNE4）的表达均有所下调。 结论 阿司咪唑对斑马鱼胚胎的心脏毒性具有一定的剂量依赖性，且该心脏毒性的作用机制很可能是首先干扰心脏中各离子通道相关基因的表达，进而干扰离子通道的正常功能，最终引发心脏毒性。

[关键词] 阿司咪唑；斑马鱼；心脏毒性；分子机制

[中图分类号] R965 [文献标识码] A [文章编号] 1673-9701（2017）24-0041-05

[Abstract] Objective To study the cardiotoxicity of astemizole and its molecular mechanism by using zebrafish as the experimental animals. Methods Twenty-four post fertilization AB-zebrafish were selected as experimental animals and exposed to different concentrations of astemizole solution. After 24 h treatment， the mortality was calculated and the morphological changes of the heart were observed under the microscope. The heart rate changes were recorded. RT-qPCR technique was used to detect changes in cardiac function-related gene expression. Results As the concentration of astemizole was increased， zebrafish embryos gradually showed death， heart sac edema， heart deformity， heart rate decline and so on； the expression of seven cardiac function-related genes（CAV1.3a， CAV1.3b， HCN2， HCN4， KCNH2a， KCNQ1 and KCNE4） were down-regulated. Conclusion Astemizole has a dose-dependent effect on the cardiac toxicity of zebrafish embryos. The mechanism of cardiotoxicity is likely to first interfere with the expression of each ion channel-related genes in the heart， and then interfere with the normal function of the ion channel， ultimately leading to cardiotoxicity.

[Key words] Astemizole； Zebrafish； Cardiotoxicity； Molecular mechanism

近些年来，新药研发的投入虽有大幅提高，但成功上市的新药并没有相应的增多。主要原因即是在新药研发的不同阶段，特别是临床前试验阶段，许多化合物因安全性和有效性等因素而被迫中止研发；其中心脏毒性是最为常见的因素之一[1]。建立快速测定心脏毒性的筛选模型对药物进行早期预测评价，将有助于降低新药研发的后期成本、提高新药研发的成功率，因此对于降低新药研发的风险及上市药物的再评价等均具有非常重要的意义。

斑马鱼（zebrafish）作为一种新兴的脊椎动物模式生物，其个体较小、易于饲养、体外受精、体外发育、胚胎及幼鱼通体透明[2]，便于直接活体观测心血管系统的表型改变；……