抗坏血酸联合全反式维甲酸建立斑马鱼新药早期评价模型的研究
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摘要
目的以抗坏血酸和全反式维甲酸为工具药,建立斑马鱼药物早期评价模型,为药物毒性的早期筛查提供实验方法。方法以斑马鱼胚胎培养用水为空白对照,二甲基亚砜为助溶剂,抗坏血酸为阴性药物,全反式维甲酸为阳性药物,对脱膜斑马鱼受精卵连续染毒5d后,检测受精卵和幼鱼的死亡率,及幼鱼自由活动度、全身组织器官形态发育和骨骼发育情况。结果 0.5%二甲基亚砜和10 mg·L~(-1)抗坏血酸对斑马鱼致死作用较弱,且无明显发育毒性作用,可分别用作常规助溶剂和质量控制标志药;全反式维甲酸随给药浓度升高,致畸效应加强,很好地呈现了自主活动缓慢到丧失、形态发育轻微异常到全身各组织器官严重畸形、骨骼矿化正常到抑制的全效应谱,为药物毒性的评价提供了很好的参照,其中7.5μg·L~(-1)可作为药物评价中阳性对照选用浓度。结论用抗坏血酸和全反式维甲酸建立的斑马鱼模型设计合理,可用于药物毒性的早期评价。
OBJECTIVE To establish a zebrafish model of drug early evaluation, using ascorbic acid and all-trans retinoic acid as tools, so as to provide a test method for early screening of drug toxicity.METHODS The zebrafish embryos were incubated for 5 days with water as the blank control, dimethyl sulfoxide as solvent, ascorbic acid(AA) as negative drug, all-trans retinoic acid(ATRA) as positive drug.Detected the mortality, movement, organs and skeleton morphogenesis of fertilized eggs and juveniles.RESULTS 0.5% DMSO and 10 mg·L~(-1) AA had weak lethal effect on zebrafish, and no obvious developmental toxicity, which could be used as cosolvent and quality control drug, respectively.ATRA had a strengthened teratogenic effect with the concentration increased, and showed a full-effect spectrum as follows: normal, slow and losed spontaneous activities, slight to severe tissues and organs malformations, vertebra mineralization inhibition.It provided a good reference for the evaluation of drug toxicity, and ATRA 7.5 μg·L~(-1) could be selected as the positive control drug evaluated concentration.CONCLUSION The zebrafish model designed with AA and ATRA reasonably can be used for early evaluation of drug toxicity.
OBJECTIVE To establish a zebrafish model of drug early evaluation, using ascorbic acid and all-trans retinoic acid as tools, so as to provide a test method for early screening of drug toxicity.METHODS The zebrafish embryos were incubated for 5 days with water as the blank control, dimethyl sulfoxide as solvent, ascorbic acid(AA) as negative drug, all-trans retinoic acid(ATRA) as positive drug.Detected the mortality, movement, organs and skeleton morphogenesis of fertilized eggs and juveniles.RESULTS 0.5% DMSO and 10 mg·L~(-1) AA had weak lethal effect on zebrafish, and no obvious developmental toxicity, which could be used as cosolvent and quality control drug, respectively.ATRA had a strengthened teratogenic effect with the concentration increased, and showed a full-effect spectrum as follows: normal, slow and losed spontaneous activities, slight to severe tissues and organs malformations, vertebra mineralization inhibition.It provided a good reference for the evaluation of drug toxicity, and ATRA 7.5 μg·L~(-1) could be selected as the positive control drug evaluated concentration.CONCLUSION The zebrafish model designed with AA and ATRA reasonably can be used for early evaluation of drug toxicity.
引文
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[2]XIANG Q Q,XU B F,DING Y L,et al.Oxidative stress response induced by butachlor in zebrafish embryo/larvae:the protective effect of vitamin C[J].Bull Environ Contam Toxicol,2018,100(2):208-215.
[3]DING H,ZHOU X H,CAI A L.Basic fibroblast growth factor expression in serum and amniotic fluid of all-trans retinoic acid induced bone deformity[J].BME Clin Med(生物医学工程与临床),2014,18(2):178-180.
[4]BRANNEN K C,PANZICA-KELLY J M,DANBERRY T L,et al.Development of a zebrafish embryo teratogenicity assay and quantitative prediction model[J].Birth Defects Res B Dev Reprod Toxicol,2010,89(1):66-77.
[5]NOHALEZ A,MARTINEZ C A,PARRILLA I,et al.Exogenous ascorbic acid enhances vitrification survival of porcine in vitro-developed blastocysts but fails to improve the in vitro embryo production outcomes[J].Theriogenology,2018,113:113-119.