中国药物警戒 ›› 2015, Vol. 12 ›› Issue (12): 711-716.

• 益母草提取方式与毒性研究专栏 • 上一篇    下一篇

水沉对益母草醇提物的小鼠急性毒性及毒靶器官的影响

孙玲1, 李晓宇2, 孙晓倩3, 窦立雯3, 石亮3, 赵红4, 孙蓉2*   

  1. 1山东省药学科学院,山东 济南 250101;
    2山东省中医药研究院,山东 济南 250014;
    3山东中医药大学,山东 济南 250355;
    4中国人民解放军海军总医院,北京 10004
  • 收稿日期:2016-02-02 修回日期:2016-02-02 出版日期:2015-12-08 发布日期:2016-02-02
  • 通讯作者: 孙玲,女,硕士,主任药师,药物分析与新药研发。E-mail:sunl_70@126.com
  • 作者简介:孙玲,女,药学硕士,主任药师,药物分析与新药研发。
  • 基金资助:
    国家自然科学基金项目(81450021); 山东省自主创新和成果转化项目(2014ZZCX02104)

Influence of Water Precipitating on Acute Toxicity and Toxicity Target Organs of Alcohol Extracts from Herba Leonuri in Mice

SUN Ling1, LI Xiao-yu2, SUN Xiao-qian3, DOU Li-wen3, SHI Liang3, ZHAO Hong4, SUN Rong2, *   

  1. 1Shandong Academy of Pharmaceutical Sciences, Shandong Ji'nan 250101, China;
    2Shandong Academy of Chinese Medicine, Shandong Ji'nan 250014, China;
    3Shandong University of Traditional Chinese Medicine, Shandong Ji'nan 250014, China;
    4PLA Navy General Hospital, Beijing 100048, China
  • Received:2016-02-02 Revised:2016-02-02 Online:2015-12-08 Published:2016-02-02

摘要: 目的比较水沉工艺对益母草醇提物小鼠急性毒性的影响及其毒性靶器官损伤程度的差异性。方法运用紫外分光光度法测定不同样品中益母草总生物碱的含量,急性毒性实验法对其急性毒性进行比较,记录其毒性症状、累积死亡数和小鼠体重变化,并于药后7天,取血,检测血液生化指标,取肝脏、肾脏称重,并计算脏器系数和进行病理组织学检查。结果益母草醇提物、醇提水沉物中总生物碱含量分别为0.82%和0.65%,益母草醇提物LD50按含生药量计算为118.68 g·kg-1,相当于临床70 kg人日用量的276.9倍;95%可信限为112.84~124.89 g·kg-1,相当于临床70 kg人日用量的263.3~291.4倍。益母草醇提水沉物的MLD为433.20 g·kg-1,相当于临床70 kg人日用量的1 010.8倍。益母草醇提和醇提水沉物均可导致小鼠BUN、Cr水平明显升高(P<0.001),肾脏系数升高,经统计学检验与正常对照组比较呈现不同程度的显著性差异;醇提物与醇提水沉物比较,醇提1~4组小鼠BUN、Cr水平升高,呈现不同程度的显著性差异,肾脏系数明显升高,经统计学检验,呈现不同程度的显著性差异;益母草醇提1~2组和醇提水沉物均可导致ALT、AST水平升高,经统计学检验与正常对照组比较呈现不同程度的差异性,醇提1组可导致肝脏系数升高,有非常显著性差异(P<0.01);醇提物与醇提水沉物比较,醇提1~2组的ALT、AST水平和肝脏系数升高,呈现不同程度的显著性差异。肝、肾病理表明,小鼠肝、肾组织未见明显病理改变。结论急性毒性试验显示益母草提取物的毒性靶器官主要为肾脏、肝脏,表现为BUN、Cr水平的显著增高和ALT、AST水平的增高和肝、肾脏系数的增加,其中肾脏毒性重于肝脏,益母草醇提物毒性大于醇提水沉物,结合益母草醇提物和醇提水沉的总碱含量数据,提示益母草提取物的毒性与总生物碱的含量呈现一定相关性,水沉工艺后总生物碱的含量和毒性显著降低,提示水沉除去的脂溶性杂质也可能是益母草的毒性成分,其确切毒性物质基础还有待进一步深入研究。

关键词: 益母草, 水沉, 醇提物, 急性毒性, 靶器官

Abstract: ObjectiveTo study the influence of water precipitating on alcohol extracts from Herba Leonuri for acute toxicity and damage degree of toxicity target organs in mice. MethodsUV spectrophotometry was used to measure the content of total alkaloids and the method of acute toxicity was used to compare the difference of acute toxicity between different alcohol extraction processes, and its toxic symptoms, cumulative mortality and weight change in mice were recorded. Seven days after administration, blood was taken, the contents of serum biochemical indicators were detected, livers and kidneys were taken for organ coefficient. ResultsThe contents of total alkaloids in the alcohol extracts and water precipitated alcohol extracts of Herba Leonuri were separately 0.82% and 0.65%. The LD50 of alcohol extracts from Herba Leonuri is 118.68 g·kg-1, which is equal to 276.9 times of human daily dried medicinal herb dose. The 95% confidence interval is 112.84~124.89 g·kg-1, which is equal to 263.3~291.4 times of human daily dried medicinal herb dose. The MLD of water precipitated alcohol extracts from Herba Leonuri is 433.20 g·kg-1, which is equal to 1 010.8 times of human daily dried medicinal herb dose. The alcohol extracts and water precipitated alcohol extracts of Herba Leonuri all could lead to significant increase (P <0.001) of mice's BUN and Cr levels. Compared with normal control group, kidney coefficient increased significantly, and the statistical tests showed varying degree difference. Compared with water precipitated alcohol extracts of Herba Leonuri, the BUN and Cr levels increased in group 1~4 of alcohol extracts, and the statistical tests showed varying degree difference. Kidney coefficient increased significantly, and the statistical tests showed varying degree difference. The group 1~2 of alcohol extracts and water precipitated alcohol extracts from Herba Leonuri could result in increased ALT and AST. Compared with normal control group, the statistical tests showed the varying degree difference. Group 1 of alcohol extracts could lead to liver coefficient increase which showed very significant difference (P <0.01). Compared with water precipitated alcohol extracts of Herba Leonuri, the ALT and AST levels and liver coefficient in group 1~2 of alcohol extracts increased. The statistical tests showed the varying degree difference. Liver and kidney pathology showed that there were no obvious pathological changes in liver and kidney tissues in mice. ConclusionAcute toxicity test showed that the toxicity target organs of Herba Leonuri were mainly kidney and liver. It proved that BUN and Cr levels significantly increased, ALT and AST levels increased, liver and kidney coefficients increased. The toxicity of kidney was higher than that of liver, and the toxicity of alcohol extract is higher than that of water precipitated alcohol extract. Combined with assay data of alcohol extract and water precipitated alcohol extract from Herba Leonuri, it is suggested that the toxicity of Herba Leonuri's extracts and total alkaloids content showed a certain correlation. Alcohol extracts with water precipitating can reduce toxicity significantly and the content of total alkaloids. Maybe water precipitating can remove the lipid soluble impurities which is also the toxic components of Herba Leonuri. But the specific toxic substances need to be further studied.

Key words: Herba Leonuri, water precipitating, alcohol extract, acute toxicity, toxicity target organ

中图分类号: