[关键词]
[摘要]
目的 研究牛黄镇惊丸对斑马鱼胚胎发育的影响。方法 以斑马鱼胚胎为实验对象,体视光学显微镜下观察牛黄镇惊丸不同给药浓度(40、200、400、800、1 200 μg/mL)时斑马鱼胚胎的发育情况,包括胚胎致畸、致死检测。结果 给药浓度不高于200 μg/mL时,未见胚胎发育异常。浓度高于200 μg/mL(约相当于成人用量30倍,远超出正常有效剂量)时对胚胎发育具有一定的毒性,胚胎出现头和眼发育较小,瞳孔无色;心脏畸形、心率较弱、血池充血-回心血流不畅;腹部透明度较差、尾干扭曲,卵黄囊延伸结构变粗,脊索变细,无正常胚动,体长变短;撤药后畸形表型未见恢复,且毒性呈浓度相关性;半数致畸浓度(TD50,受精后3 d)约373 μg/mL,半数致死浓度(LD50)约为485 μg/mL。浓度为1 200 μg/mL时,胚胎在受精后1 d之内全部死亡。结论 牛黄镇惊丸给药浓度大于200 μg/mL时影响斑马鱼胚胎发育。其毒性作用发生在胚胎器官形成期,主要对胚胎的神经系统、心血管系统和骨骼等发育有影响。
[Key word]
[Abstract]
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.
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[基金项目]
国家十二五“重大新药创制”专项课题(2014ZX09304307-002)