斑马鱼模型评价5种中药肝脏毒性

Zebrafish as an Animal Model for Assessing Hepatotoxicity of Five Chinese Medicines

目的研究壮骨关节丸、益肾乌发口服液、牛黄解毒片、雷公藤多苷和柴胡皂苷D对斑马鱼的肝脏毒性,并验证斑马鱼模型用于中药肝毒性快速评价的实用性,为临床用药提供依据。方法受精后3 d(3dpf)斑马鱼分别用不同浓度的中药处理2 d后,在显微镜下拍照分析,观察斑马鱼肝变性、肝脏形态学变化和卵黄囊吸收情况。结果与对照组相比,壮骨关节丸处理后的斑马鱼肝脏肿大,肝脏变性明显,卵黄囊吸收延迟;益肾乌发口服液致肝脏萎缩,肝脏变性明显,卵黄囊吸收显著延迟;牛黄解毒片致肝脏萎缩,卵黄囊吸收显著延迟;雷公藤多苷和柴胡皂苷D致肝脏肿大,斑马鱼卵黄囊吸收均显著延迟。结论壮骨关节丸、益肾乌发口服液、牛黄解毒片、雷公藤多苷和柴胡皂苷D对斑马鱼幼鱼具有不同程度的肝脏毒性。斑马鱼模型评价中药肝脏毒性具有较好的预测性。

Objective To determine the hepatotoxicity of 5 known mammalian hepatotoxic drugs（ Zhuangguguanjie[ZGGJ]pill,Yi Shen Wu Fa oral liquid,Niuhuang Jiedu Pian,Tripterygium Glycosides [TWP],saikosaponin d） in larval zebrafish and to validate the practicability of zebrafish model for rapid assessment of drug hepatotoxicity. Method Zebrafish at 3 dpf（ days past fertilization） were treated with Chinese medicines at series of concentration for 2 days. At the end of treatment,zebrafish from each group were randomly selected and images were acquired under a fluorescent stereomicroscope and quantitatively assessed in larval zebrafish using three specific phenotypic endpoints of hepatotoxicity： liver degeneration,changes in liver size and yolk sac retention.Zebrafish liver degeneration was originally screened visually, quantified using an image-based morphometric analysis. Result ZCGJ pill induced liver degeneration and reduced liver size in larval zebrafish,Yi Shen Wu Fa oral liquid induced liver degeneration and increased liver size in larval zebrafish,Niuhuang Jiedu Pian reduced liver size in larval zebrafish,TWP and saikosaponin d increased liver size in larval zebrafish. All the tested mammalian hepatotoxic drugs delayed yolk sac absorption in larval zebrafish. Conclusion Chinese medicines could induce the liver damage in larval zebrafish. Zebrafish phenotypic assay is a highly predictive animal model for rapidly in vivo assessment of compound hepatotoxicity.