摘要
目的:采用斑马鱼模型高效筛选壮骨关节丸中有毒性的药味。方法:将受精后1天(1 day post fertilization,1dpf)的斑马鱼胚胎暴露于壮骨关节丸及方中12种药味不同浓度水煎液和醇提液中,观察记录给药后2-6 dpf或2-8 dpf的死亡数量,于3 dpf显微检视斑马鱼幼鱼状态,SPSS软件计算6 dpf的斑马鱼半数致死浓度LC50。结果:斑马鱼暴露于壮骨关节丸及补骨脂、续断、乳香没药、独活和木香的水、醇提取液,以及鸡血藤和狗脊的醇提液毒性较大,致鱼卵黄囊肿大、变形或变黑,提示与肝毒相关,6 dpf鱼的LC50为33.2-392.7μg·m L-1(已折算为生药量,下同);淫羊藿水、醇提取液均在1 500μg·m L-1致鱼卵黄囊肿大、变黑,LC50为1 100-1 350μg·m L-1,有一定毒性;鸡血藤和狗脊的水煎液,以及骨碎补、桑寄生和熟地黄的水、醇提取液未致鱼脏器明显改变,LC50大于1 500μg·m L-1,故安全性较好。结论:斑马鱼模型高效辨识了壮骨关节丸不同药味毒性差异,可实现复杂中药中潜在毒性物质的在体、实时、动态、规模化的高效筛选。
This study aimed at efficiently screening out the toxic herbal medicines in Zhuang Gu Guan Jie pill（ZGGJ） based on the zebrafish model. Zebrafish embryos at 1 day post fertilization（dpf） were administered by ZGGJ and the water decoction（WD） and ethanol extract（EE） of 12 herbal drugs in it with various concentrations. The number of embryo deaths after the administration was recorded at 2-6 dpf and 2-8 dpf, respectively. Embryonic micro-morphology of zebrafish（3 dpf） was observed, LC50 value at 6 dpf was calculated by SPSS16.0. As a result, both the WD and EE of ZGGJ, fructus psoraleae, radix dipsaci, olibanum, myrrh, radix angelicae pubescentis and aucklandiae radix, and the EE of Caulis Spatholobo and Rhizoma Cibotii presented high organ toxicity to the juvenile zebrafish, featuring yolk sac swelling, deforming, blackening, indicating hepatotoxicity in the zebrafish. Their LC50 value at 6 dpf was from 33.2 μg·m L-1 to 392.7 μg·m L-1（LD50 value was converted to crude drug. The same bellow.）. The WD and EE of Epimedii Folium at 1,500 μg·m L-1 led to yolk sac swelling and blackening. The LC50 at 6 dpf varied at 1,100-1,350 μg·m L-1, showing a certain toxicity of Epimedii Folium. The WD of Caulis Spatholobo and Cibotii Rhizoma and both WD and EE of Rhizoma drynariae, taxilli herba and prepare-d rehmanniae radix didn＇t change the toxicity to the organs in zebrafish. LC50 value were over 1,500 μg·m L1, presenting favorable safety. In conclusion, it was demonstrated that the zebrafish model efficiently identified the toxicity of different herbal medicines from ZGGJ and efficiently screened out the potential toxicants in vivo with the advantages of real-time, dynamic and large scale for traditional Chinese medical compounds.
出处
《世界科学技术-中医药现代化》
CSCD
2017年第1期133-141,共9页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
国家自然科学基金委面上项目(81573833):基于三维M-Act/Tox协同评价的抗骨质疏松中药筛选新方法的建立与应用
负责人:韦英杰
江苏省科技厅自然科学基金面上项目(BK20141507):基于斑马鱼M-Act/Tox一体化评价的中药抗骨质疏松筛选新方法与应用
负责人:韦英杰
江苏省人才工作领导小组办公室2014年度江苏省第四期"333工程"科研资助项目(RA2014348):基于斑马鱼毒/效同步评价的抗骨质疏松筛选中药新技术与应用
负责人:韦英杰
江苏省人力资源和社会保障厅江苏省第十批"六大人才高峰"(2013-YY006):以二维斑马鱼模型联合色谱联用技术高效筛选中药抗骨质疏松体内外活性成分
负责人:韦英杰
关键词
壮骨关节丸
斑马鱼
毒性药味
肝毒性
Zhuang Gu Guan Jie pill
zebrafish
toxicity of herbal medicine
hepatotoxicity
