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纸质出版日期:2018
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罗隽, 梅雪, 夏青, 等. 醋甘遂不同提取物对斑马鱼幼鱼的毒性[J]. 中国实验方剂学杂志, 2018,24(4):160-166.
LUO Jun, MEI Xue, XIA Qing, et al. Toxicity of Different Vinegar-preparing Kansui Radix Extracts on Zebrafish Larvae[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(4): 160-166.
罗隽, 梅雪, 夏青, 等. 醋甘遂不同提取物对斑马鱼幼鱼的毒性[J]. 中国实验方剂学杂志, 2018,24(4):160-166. DOI: 10.13422/j.cnki.syfjx.2018040160.
LUO Jun, MEI Xue, XIA Qing, et al. Toxicity of Different Vinegar-preparing Kansui Radix Extracts on Zebrafish Larvae[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(4): 160-166. DOI: 10.13422/j.cnki.syfjx.2018040160.
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3 d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,肝脏颜色变暗(P<0.01);肝脏经苏木精-伊红染色后,在石油醚提取部位0.054 mg·L-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。其他系统毒性特征不明显,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大,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 investigate the toxicity and toxicity reversibility of non-lethal dose of vinegar-preparing Kansui Radix on zebrafish larvae
and validate the feasibility of zebrafish model for rapid assessment of drug toxicity. Method: Samples were continuously extracted with soxhlet extractor in different polar solvents to prepare different extracts. Healthy adult zebrafish were paired for spawning. The 72 hour-post-fertilization(hpf) zebrafish larvae were chosen as models to investigate their mortality rate after 72 h treatment with different extracts of processed Kansui. The toxicity and reversibility of the most toxic extraction part was evaluated on zebrafish cardiovascular system
liver
nervous system
gastrointestinal system
kidneys and other organs and systems. Result: The order of toxicities for different extraction parts of processed Kansui Radix on zebrafish larvae was as follows: petroleum ether extract > ethyl acetate extract > dichloromethane extract > ethanol extract. In the petroleum ether extract treated group
the liver intensity of the zebrafish larvae was significantly changed with the drug concentration
and the color the liver became dark as compared with blank group (P<0.01). Minor liver injury was found in the 0.054 mg·L-1 petroleum ether extract group by hematoxylin-eosin staining
but it was significantly improved in the recovery group. The residual was decreased significantly in 0.012
0.036
0.054 mg·L-1 petroleum ether extract groups as compared to blank control group and 0.004 mg·L-1 petroleum ether extract group (P<0.01)
and their gastrointestinal tract was promoted to empty. Other systems had no obvious toxicity characteristics
with no statistically significant differences between the groups. Conclusion: In the petroleum ether
dichloromethane
ethyl acetate and ethanol extracts of vinegar-preparing Kansui
petroleum ether extract had the greatest toxicity
mainly in hepatotoxicity and gastrointestinal irritation on zebrafish larvae. Hepatotoxicity caused liver degeneration and showed high reversibility. What's more
zebrafish as a reliable mammal model for screening acute toxic agents has good application prospects.
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