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

Zebrafish Model of New Drug Early Evaluation Established by Ascorbic Acid and All-trans Retinoic Acid

关键词:  抗坏血酸  全反式维甲酸  斑马鱼  药物毒性检测

Key words:  ascorbic acid  all-trans retinoic acid  zebrafish  drug evaluation

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5 d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7.5 μg·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。

Abstract:

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.