张楷承+曹雨诞+姚芳+张丽+丁安伟

[摘要] 采用模式生物斑马鱼的胚胎评价京大戟醋制前后各提取物急性毒性，同时以大戟二烯醇为对照品测定了各提取物的总萜含量。选取24 h发育正常的斑马鱼胚胎，每个提取物设8个浓度和一个空白对照组，并观察给药后96 h斑马鱼胚胎的发育和死亡情况，计算不同样品对斑马鱼胚胎的半数致死浓度（LC₅₀）。结果显示对于斑马鱼胚胎，京大戟醋制前后各提取物均有急性毒性，与生品相比，醋制后毒性显著降低。不同提取方式中，醇提物毒性大于水提物；不同极性部位中毒性大小依次为石油醚>二氯甲烷>乙酸乙酯>正丁醇、剩余部位。结合萜类成分含量测定结果可推测萜类成分为京大戟主要毒性成分，且其毒性大小与萜类成分的含量呈正相关。表明斑马鱼胚胎模型适用于京大戟毒性评价，为进一步认识与评价京大戟的毒性成分及醋制减毒的作用机制提供合适的研究方法以及理论依据。

[关键词] 京大戟； 醋制； 斑马鱼胚胎； 急性毒性； 总萜含量

[Abstract] The embryos of model organism zebrafish were used to evaluate the acute toxicity of the extracts of Euphorbiae Pekinensis Radix and vinegar-processing Euphorbiae Pekinensis Radix， and the total terpene content of each extract was determined by using euphol as the reference standards. Twenty-four h normally developed zebrafish embryos were chosen， and 8 concentrations were adopted for each extract. Then the growth and death of zebrafish embryos were observed at 96 h after administration， and median lethal concentrations （LC₅₀） of the different samples on zebrafish embryos were calculated. The results showed that all of the extracts （before and after vinegar processing） had acute toxicity on zebrafish embryos. The toxicity of vinegar-processing Euphorbiae Pekinensis Radix was significantly lower than that of crude Euphorbiae Pekinensis Radix. Among different extraction methods， ethanol extract was more poisonous than water extract； in different polarity fractions， the toxicity was in the following order： petroleum ether>dichloromethane>ethyl acetate>n-butyl alcohol and remaining part. Combined with the results of the determination of terpene components， it can be concluded that the terpenoids are the main toxic components of Euphorbiae Pekinensis Radix， positively correlated with toxicity degree. It indicates that the zebrafish embryo model is appropriate for the toxicity evaluation of Euphorbiae Pekinensis Radix and provides appropriate research methods and theoretical basis for the further study of the toxic components and the mechanism of reducing toxicity.

[Key words] Euphorbiae Pekinensis Radix； processing with vinegar； zebrafish； acute toxicity； content of total terpene

斑馬鱼Barchydanio rerio，为鲤科Danio属，最早产于孟加拉、印度等地。因其与人类基因具有高度同源性，在蛋白质水平上其关键部位的同源性几乎是100%，且各个器官如肝、肠、肾等都与人类极为相似，故可用斑马鱼模拟人类很多疾病[1-2]，作为一种新兴的模式生物，斑马鱼有着低成本、易饲养、易于给药与观察、实验动物数量多、重复性好等优点，近年来已经发展成为全球公认的医学和毒理学动物模型 [3-5]，且斑马鱼胚胎毒性研究已经延伸到了中药领域，并取得了喜人成果[6-9]。