Validation of zebrafish embryo toxicity evaluation method / 中国药理学与毒理学杂志

ABSTRACT

OBJECTIVE In this study,nine different levels of known teratogenic embryos toxic com-pounds (aceta minophen,methi mazole,indo metacin,methotrexate,ascorbic acid,isoniazid,penicillin G,saccharin ,5-fluorouracil )were used to validate the established evaluation method of zebrafish e mbryo develop mental toxicity in our laboratory via zebrafish e mbryonic develop ment toxicity tests. METHODS By water bath method,zebrafish embryos of post-fertilization 2 h (2 hpf)were exposed to different concentrations of compounds.Observe and record the nu mber of malformations and dead e mbryo in 24,48,72,144 hpf.After three parallel experiments,get EC50 ,LC50 through software and calculate the teratogenic indices (TI =LC50 /EC50 )of the positive drug and recorded it as teratogens.If there is no teratogenic index recorded it as non-teratogens.Co mpare the results with the existing animal experiments and clinical trials to obtain the sensitivity and specificity.RESULTS Based on the results, exposure to aceta minophen (TI =2.07),methi mazole (TI =2.91 ),indo methacin (TI =1 .67),5-flu-orouracil (TI =8.31 ),methotrexate (TI =1 .31 ),showed various levels of teratogenic effects,and the corresponding TI′s were calculated.Methi mazole appeared more obvious skeletal deformities;indo m-ethacin toxicity is mainly reflected in abnormal heart,blood circulation and other endpoints;5-FU at lower doses that result in higher mortality rates,but also took place over the lacking of pig mentation;aceta min-ophen for the heart,liver and other target organ toxicity,pericardial cyst;methotrexate appears signifi-cant so mites,skeletal deformity.Since the IT′s for ascorbic acid,isoniazid,penicillin G,saccharin can-not be calculated,these compounds are considered to show negative results,indicating that the data fro m the nine compounds in zebrafish test are consistent with traditional animal experiments and clinical information,the sensitivity of 100% and specificity of 100%.CONCLUSION Though validation test,it has been de monstrated that the established zebrafish embryos develop mental toxicity evaluation method can be applied to early screening of drug toxicity.