基于斑马鱼幼鱼模型的甘遂毒性评价

Toxicity evaluation of Radix Kansui on zebrafish larvae

[目的]以模式生物斑马鱼幼鱼为模型,评价甘遂不同溶剂连续提取物毒性差异及毒性特征。[方法]采用索氏提取法获得甘遂不同溶剂连续提取物。通过预实验,确定各提取物72 h最大不致死浓度和最小全致死浓度。在此区间设定6~8个药物浓度,建立致死曲线,计算半数致死浓度(LC50)。以毒性最大提取物的10%致死浓度(LC10)、最小致死浓度(LC1)、1/3 LC1和1/10 LC1为给药浓度,同时设立空白组和溶剂对照组,连续给药72 h,观察甘遂毒性特征。停药后将幼鱼置于养殖水中恢复48 h,观察毒性可逆性。[结果]甘遂不同提取部位毒性顺序为石油醚>二氯甲烷>乙酸乙酯>乙醇提取物。甘遂的主要毒性为肝毒性和胃肠道刺激性。肝毒性既可造成肝肿大,又可导致肝变性,肝肿大不可逆,而肝变性具有可逆性。[结论]本实验首次利用单个模型以相同浓度单次给药,同时评价了甘遂对心脏、肝脏、肾脏、神经系统和胃肠道等脏器系统的毒性作用及可逆性。实验结果证实了甘遂的毒性集中于石油醚提取部位,并可对肝脏和胃肠道造成实质性损伤。本实验为斑马鱼模型在中药毒性评价中的应用提供了参考。

[Objective] To evaluate the toxicity of Radix Kansui extracted with different solvent on zebrafish larvae.[Methods] The Radix Kansui was extracted with different solvent continuously by Soxhlet extraction method.The maximal non-lethal concentration and minimum total lethal concentration of each extract for 72 h were determined by pre-experiment.The median lethal concentration（LC50）was calculated by lethal curve which was established with 6~8 drug concentrations that are set in accordance with pre-experiment.10%lethal concentration（LC10）,the minimum lethal concentration（LC1）,1/3 LC1 and 1/10 LC1 of the most toxic extract were set up as concentrations of toxicity characteristic observation experiment.The control group and the solvent control group were set up at the same time.The toxicity characteristics of zebrafish larvae were observed after 72 h exposure with different concentrations.The toxicity reversibility was observed after the larvae living in fish water 48 h.[Results] The toxicity order of Radix Kansui extracts was petroleum ether extract dichloromethane extract ethyl acetate extract ethanol extract.The toxicity of Radix Kansui is mainly characterized as liver toxicity and gastrointestinal irritation.Liver toxicity was showed as hepatomegaly and liver degeneration.Hepatomegaly is irreversible,but liver degeneration is reversible.[Conclusion] We simultaneously evaluated the toxic effects and reversibility of Radix Kansui on heart,liver,kidney,nervous system and gastrointestinal tract using one model with one administration for the first time.The results confirmed that the toxicity of Radix Kansui was mainly derived from petroleum ether extracts.Radix Kansui can cause substantial damage to the liver and gastrointestinal tract.This study provides a reference for the application of zebrafish in the evaluation of traditional Chinese medicine toxicity.