目的 研究牛黃鎮(zhèn)驚丸對斑馬魚胚胎發(fā)育的影響。方法 以斑馬魚胚胎為實驗對象，體視光學顯微鏡下觀察牛黃鎮(zhèn)驚丸不同給藥濃度（40、200、400、800、1 200 μg/mL）時斑馬魚胚胎的發(fā)育情況，包括胚胎致畸、致死檢測。結(jié)果 給藥濃度不高于200 μg/mL時，未見胚胎發(fā)育異常。濃度高于200 μg/mL（約相當于成人用量30倍，遠超出正常有效劑量）時對胚胎發(fā)育具有一定的毒性，胚胎出現(xiàn)頭和眼發(fā)育較小，瞳孔無色；心臟畸形、心率較弱、血池充血-回心血流不暢；腹部透明度較差、尾干扭曲，卵黃囊延伸結(jié)構(gòu)變粗，脊索變細，無正常胚動，體長變短；撤藥后畸形表型未見恢復(fù)，且毒性呈濃度相關(guān)性；半數(shù)致畸濃度（TD₅₀，受精后3 d）約373 μg/mL，半數(shù)致死濃度（LD₅₀）約為485 μg/mL。濃度為1 200 μg/mL時，胚胎在受精后1 d之內(nèi)全部死亡。結(jié)論 牛黃鎮(zhèn)驚丸給藥濃度大于200 μg/mL時影響斑馬魚胚胎發(fā)育。其毒性作用發(fā)生在胚胎器官形成期，主要對胚胎的神經(jīng)系統(tǒng)、心血管系統(tǒng)和骨骼等發(fā)育有影響。

Objective To study the toxic effect of Niuhuang Zhenjing Pill on zebrafish embryo development.Methods Zebrafish embryo was used to investigate the development situation under stereo microscope at different drug concentration (40, 200, 400, 800, 1 200 μg/mL) including embryo teratogenic and lethal tests.Results No abnormal embryo development was found when the concentration was not greater than 200 μg/mL (Approximately equal to 30 times the normal adult dosage, exceeding the normal effective dose). While, there was toxicity to embryo development at the concentration greater than 200 μg/mL. The head and eyes of the embryo were smaller, and the pupils were colorless. Zebrafish embryonic heart malformation was observed, heart rate was weak, blood pool congestion happened and back blood flow was not smooth. The abdominal surface of the embryo was poor in transparency, the tail was twisted, the yolk sac elongation structure became thicker, the notochord became thin, there was no normal embryo movement, and the body length became shorter. After the withdrawal, the abnormal phenotype was not recovered, and severity of the abnormal phenotype was concentration-dependent. It was also observed that all embryos died within 1 d after fertilization at the concentration of 1 200 μg/mL.Conclusion The drug could influence the development of zebrafish embryos when the concentration was higher than 200 μg/mL. Its toxic effect occurs during embryonic organogenesis. And it mainly affects the development of embryonic nervous system, cardiovascular system and bone.

國家十二五“重大新藥創(chuàng)制”專項課題（2014ZX09304307-002）