基于模式生物斑马鱼的款冬叶肝肾毒性比较研究
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摘要
目的采用斑马鱼模型比较款冬叶、花的肝脏、肾脏毒性,为款冬叶的资源利用提供实验依据。方法取3 dpf(day post fertilization)健康AB系及转基因系斑马鱼,将款冬叶、款冬花以及款冬叶、花分别与紫菀配伍后的4个样品溶于斑马鱼培养用水,分别以0.5、1.0、1.5 mg/mL给药,以培养用水为对照,处理72 h后观察各组样品对斑马鱼肝、肾形态及各指标的影响,以斑马鱼肝脏的荧光面积和肝脏丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)水平评价药物对肝脏的毒性,以斑马鱼表型变化结合培养液蛋白量评价药物的肾脏毒性。结果与对照组比较,各给药组对斑马鱼肝脏生化指标无显著影响,但在1.5 mg/mL剂量下,款冬花及款冬花+紫菀组斑马鱼的肝脏荧光面积减小。各药物处理组斑马鱼表型及培养液蛋白量无显著差异。结论款冬花及款冬花配伍紫菀在高剂量时对肝脏产生一定的毒性,而款冬叶及款冬叶配伍紫菀未表现出明显的肝肾毒性,即款冬叶入药的安全性风险不高于款冬花。研究结果为款冬叶的资源利用奠定了基础。
Objective To compare the toxicity of the flower buds and leaves from Tussilago farfara, and provide scientific basis for the utilization of the leaves of T. farfara. Methods The 3 dpf(day post fertilization) healthy AB and transgene zebrafish were selected. The flowers, the leaves, the flowers coupled with Aster tataricus, and the leaves coupled with A. tataricus were prepared at the concentration of 0.5, 1.0, and 1.5 mg/mL. The fluorescent area of the liver and ALT and AST levels were measured after 72 h of drug treatment. For the renal toxicity assay, the morphology of zebrafish and the nutrient solution protein were also determined. ResultsCompared with the control group, there were no significant differences in liver biochemical indexes in the four drug treatment groups. However, the fluorescence area of liver decreased in the flowers and flowers coupled with A. tataricus group at the concentration of 1.5 mg/mL. No significant difference was observed in the four groups of the nephrotoxicity assay. Conclusion The flower and the flower coupled with A. tataricus showed minor hepatotoxicity at higher doses, and the leaves showed no stronger toxicity than the flower buds in comparison with the flower buds. It can provide refercences for the resource utilization of the leaves of T. farfara.
Objective To compare the toxicity of the flower buds and leaves from Tussilago farfara, and provide scientific basis for the utilization of the leaves of T. farfara. Methods The 3 dpf(day post fertilization) healthy AB and transgene zebrafish were selected. The flowers, the leaves, the flowers coupled with Aster tataricus, and the leaves coupled with A. tataricus were prepared at the concentration of 0.5, 1.0, and 1.5 mg/mL. The fluorescent area of the liver and ALT and AST levels were measured after 72 h of drug treatment. For the renal toxicity assay, the morphology of zebrafish and the nutrient solution protein were also determined. ResultsCompared with the control group, there were no significant differences in liver biochemical indexes in the four drug treatment groups. However, the fluorescence area of liver decreased in the flowers and flowers coupled with A. tataricus group at the concentration of 1.5 mg/mL. No significant difference was observed in the four groups of the nephrotoxicity assay. Conclusion The flower and the flower coupled with A. tataricus showed minor hepatotoxicity at higher doses, and the leaves showed no stronger toxicity than the flower buds in comparison with the flower buds. It can provide refercences for the resource utilization of the leaves of T. farfara.
引文
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[12]Zhi H J,Qin X M,Sun H F,et al.Metabolic fingerprinting of Tussilago farfara L.using 1H NMRspectroscopy and multivariate data analysis[J].Phytochem Anal,2012,23(5):492-501.
[13]李静,李娟,贾金萍,等.基于UHPLC-QExtractive轨道阱高分辨质谱的款冬花、叶的化学比较[J].药学学报,2018,53(3):444-452.
[14]刘可越,张铁军,高文远,等.款冬花的化学成分及药理活性研究进展[J].中国中药杂志,2006,31(22):1837-1841.
[15]吴琪珍,张朝凤,许翔鸿,等.款冬花化学成分和药理活性研究进展[J].中国野生植物资源,2015,34(2):33-36.
[16]彭蕴茹,韦英杰,丁永芳,等.基于斑马鱼模型的药物毒性研究进展与中药毒性研究新策略[J].中草药,2017,48(1):17-30.
[17]常嘉,陆亮,王庆利,等.斑马鱼在药物早期毒性筛选中的应用进展[J].中国新药杂志,2013,22(13):1500-1504.
[2]吕培霖,李成义,郑明霞.甘肃款冬花资源调查报告[J].中国现代中药,2008,10(4):42-43.
[3]支海娟.基于NMR款冬植物代谢组学研究[D].太原:山西大学,2012.
[4]Li Z Y,Zhi H J,Zhang F S,et al.Metabolomic profiling of the antitussive and expectorant plant Tussilago farfara L.by nuclear magnetic resonance spectroscopy and multivariate data analysis[J].J Pharmaceut Biomed,2013,doi:10.1016/j.jpba.2012.11.023.
[5]刘可越.紫菀散药效物质基础的研究及新药研制[D].天津:天津大学,2006.
[6]陈雪园,张如松,杨苏蓓.款冬花化学成分及药理毒理研究进展[J].亚太传统医药,2012,8(1):173-174.
[7]赵崇军,田敬欢,王金凤,等.斑马鱼在中药研究中的应用进展[J].中草药,2015,46(17):2635-2648.
[8]陈磊,刘怡,梁生旺.模式生物斑马鱼在中药研究中的应用[J].药学学报,2012,47(4):434-439.
[9]陈怡君,钟玉绪,董武,等.麝香酮对斑马鱼胚胎的发育毒性[J].中国药理学与毒理学杂志,2014,28(2):267-273.
[10]夏广清,姚慧敏,董丽红,等.灵芝多糖对斑马鱼存活、发育和衰老的影响[J].中国药理学与毒理学杂志,2014,28(4):491-496.
[11]方芳,赵杰,余林中,等.乌头碱对斑马鱼心脏毒性的初步研究[J].中药药理与临床,2012,28(2):31-33.
[12]Zhi H J,Qin X M,Sun H F,et al.Metabolic fingerprinting of Tussilago farfara L.using 1H NMRspectroscopy and multivariate data analysis[J].Phytochem Anal,2012,23(5):492-501.
[13]李静,李娟,贾金萍,等.基于UHPLC-QExtractive轨道阱高分辨质谱的款冬花、叶的化学比较[J].药学学报,2018,53(3):444-452.
[14]刘可越,张铁军,高文远,等.款冬花的化学成分及药理活性研究进展[J].中国中药杂志,2006,31(22):1837-1841.
[15]吴琪珍,张朝凤,许翔鸿,等.款冬花化学成分和药理活性研究进展[J].中国野生植物资源,2015,34(2):33-36.
[16]彭蕴茹,韦英杰,丁永芳,等.基于斑马鱼模型的药物毒性研究进展与中药毒性研究新策略[J].中草药,2017,48(1):17-30.
[17]常嘉,陆亮,王庆利,等.斑马鱼在药物早期毒性筛选中的应用进展[J].中国新药杂志,2013,22(13):1500-1504.