马兜铃酸和马兜铃内酰胺对斑马鱼胚胎毒性的比较研究

Comparative Study on Toxicity of Aristolochic Acid and Aristololactam to Zebrafish Embryos

为比较马兜铃酸和马兜铃内酰胺对斑马鱼胚胎的急性毒性,考察斑马鱼作为药物毒性成分快速筛选模型的适用性,分别用不同浓度的马兜铃酸I(AA-I)和马兜铃内酰胺I(AL-I)处理斑马鱼胚胎,观察并记录药物处理后的斑马鱼胚胎发育情况,比较二者对斑马鱼胚胎的致畸率和死亡率的影响,并对斑马鱼胚胎的器官毒性进行评价,采用RT-PCR检测斑马鱼肾脏发育相关基因wt1a/b的表达变化。结果为:胚胎的死亡率、畸形率与药物浓度和暴露时间基本呈正相关,马兜铃内酰胺实验组的死亡率和畸形率均高于同等浓度的马兜铃酸实验组。马兜铃酸与马兜铃内酰胺对斑马鱼胚胎的致畸毒性表现为尾部畸形,卵黄囊异常膨大,神经管发育缺陷,躯干弯曲、变形,心肌肿大,心率不稳,神经管缺损,色素沉积减少,后主静脉血循环减慢并逐渐衰竭,最后导致死亡。与马兜铃酸相比,马兜铃内酰胺对斑马鱼胚胎有着更强的致畸和致死毒性,毒性作用具有时间和浓度依赖性。利用斑马鱼模型评价药物毒性具有较好的预测性。

Objective: Compare the acute toxicity of aristolochic acid and aristololactam on zebrafish embryos,and investigate the applicability of zebrafish as a rapid screening model for drug toxicity components. Method: Zebrafish embryos were treated with aristolochic acid I( AA-I) and aristololactam I( AA-I) at different concentrations. Then,the development and the organ toxicity of embryos was observed and recorded. The effects of the drugs on the mortality and deformity of zebrafish embryos were compared. RT-PCR was used to detect the expression of kidney development related gene wt1 a/b in zebrafish. Result: The mortality and deformity of embryos were positively correlated with drug concentration and exposure time. The mortality and deformity rate of aristololactam group were higher than those of aristolochic acid group. The teratogenic toxicity of aristolochic acid and aristololactam on zebrafish embryos is characterized by tail deformity,abnormal enlargement of yolk sac,defect of neural tube development,bending and deforming of trunk,myocardial enlargement,heart rate instability,neural tube defect,depletion of pigmentation,slow down of blood circulation of main vein and gradual exhaustion,eventually leading to death. Conclusion: Compared with aristolochic acid,aristololactam has stronger teratogenic and lethal toxicity to zebrafish embryos,and its toxicity is time and concentration dependent. The use of zebrafish model to evaluate the toxicity of drugs has a good predictability.