论文部分内容阅读
目的:研究非致死剂量醋甘遂对斑马鱼幼鱼的毒性及毒性可逆性,并验证斑马鱼模型用于中药毒性快速评价的可行性。方法:用索氏提取器以不同极性溶剂依次提取样品,制备不同提取物。将健康斑马鱼成鱼配对产卵,以正常发育受精后3d的斑马鱼幼鱼为模型,用醋甘遂不同极性提取物处理斑马鱼幼鱼,统计给药72 h死亡率,对毒性最强的提取部位观察评价其对斑马鱼心血管系统、肝脏、神经系统、胃肠道系统、肾脏及其他脏器系统的毒性特征及可逆性。结果:醋甘遂不同溶剂提取物毒性顺序依次为石油醚提取物>二氯甲烷提取物>乙酸乙酯提取物>乙醇提取物,与空白组比较,石油醚提取部位处理后的幼鱼给药组肝区相对光强度随给药浓度的升高而降低,肝脏颜色变晴(P<0.01);肝脏经苏木精一伊红染色后。在石油醚提取部位0.054 mg~(-1)给药组中有轻微的肝损伤,在恢复组中肝损伤明显好转。与空白组和石油醚提取部位0.004 mg·L~(-1)组比较,肠道尼罗红排泄实验显示残留率在石油醚提取部位0.012,0.036,0.054 mg·L~(-1)组中显著下降(P<0.01),胃肠道被促进排空。其他系统毒件特征不明显,差异均无统计学意义。结论:醋甘遂的石油醚、二氯甲烷、乙酸乙酯和乙醇提取物中,石油醚提取物毒性最人,且主要毒性为肝毒性和胃肠道刺激性,肝毒性造成肝变性,具有可逆性。斑马鱼模型快速评价中药毒性具有较好的应用前景。
Objective: To study the toxicity and toxicity reversibility of non-lethal dose of vinegar to juvenile zebrafish and verify the feasibility of zebrafish model for rapid evaluation of toxicity of traditional Chinese medicine. Methods: The samples were extracted by Soxhlet extractor with different polar solvents to prepare different extracts. The healthy zebrafish adult fish mating spawning, zebra fish juveniles three days after normal fertilization as a model, with different polar extracts of vinegar treatment of zebrafish juvenile fish, the statistical mortality of 72 h, the toxicity of Strong extraction site observation to evaluate its zebrafish cardiovascular system, liver, nervous system, gastrointestinal system, kidney and other organ system toxicity characteristics and reversibility. Results: The toxic order of different solvent extracts of vinegar was different from petroleum ether extract> dichloromethane extract> ethyl acetate extract> ethanol extract. Compared with the blank group, the juvenile fish treated with petroleum ether extract The relative light intensity of the liver area decreased with the increase of the drug concentration, the color of the liver became clear (P <0.01), and the liver was stained with hematoxylin and eosin. In the petroleum ether extract 0.054 mg ~ (-1) administration group there is a slight liver injury in the recovery group, liver injury improved significantly. Compared with the blank group and petroleum ether extract of 0.004 mg · L ~ (-1) group, the intestinal Nile red excretion experiments showed that the residual rate in petroleum ether extraction site 0.012,0.036,0.054 mg · L -1 group Decreased significantly (P <0.01), gastrointestinal tract was promoted emptying. The characteristics of other system poisons are not obvious, the difference was not statistically significant. CONCLUSION: Among the petroleum ether, methylene chloride, ethyl acetate and ethanol extracts of vinegar, the petroleum ether extract has the highest toxicity and the main toxicity is hepatotoxicity and gastrointestinal irritation. Reversible. The rapid evaluation of zebrafish model toxicity of traditional Chinese medicine has a good prospect.