中国寄生虫学与寄生虫病杂志

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大叶桉桉叶提取物对细粒棘球绦虫原头节的体外杀虫作用

赵宇宁,张皓冰*   

  1. 中国疾病预防控制中心寄生虫病预防控制所,世界卫生组织热带病合作中心,科技部国家级热带病国际联合研究中心,卫生部寄生虫病原与媒介生物学重点实验室,上海 200025
  • 出版日期:2016-06-30 发布日期:2016-10-28

In vitro Killing Effect of Eucalyptus robusta Leaves Extract on Echinococcus granulosus Protoscolices

ZHAO Yu-ning,ZHANG Hao-bing*   

  1. National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention;WHO Collaborating Centre for Tropical Diseases;National Center for International Research on Tropical Diseases, Ministry of Science and Technology;Key Laboratory of Parasite and Vector Biology, Ministry of Health, Shanghai 200025, China
  • Online:2016-06-30 Published:2016-10-28

摘要: 目的 探索大叶桉(Eucalyptus robusta)桉叶提取物体外抗细粒棘球绦虫原头节的活性。 方法 2012年1-12月每月24日收集大叶桉成熟叶片,室内自然风干。用石油醚、二氯甲烷、乙酸乙酯和无水乙醇等4种不同极性的溶剂进行超声提取。将不同浓度的提取物与原头节孵育72 h,计算虫体死亡率,计算半数致死浓度(LC50)。 结果 不同溶剂提取物的性状和产率均不同,石油醚提取物为棕黑色油膏状,二氯甲烷、乙酸乙酯、无水乙醇提取物分别为墨绿色、乳粉色和乳白色粉末状。石油醚、二氯甲烷、乙酸乙酯、无水乙醇提取物的平均产率分别为4.4%、2.1%、2.3%和2.3%,石油醚提取物的产率最高,其5月份的产率为5.4%。作用浓度为100 μg/ml时,各月份桉叶的石油醚与二氯甲烷提取物对原头节的杀灭作用均达到100%;50 μg/ml时,石油醚提取物的杀灭作用也基本达到100%,二氯甲烷提取物次之,强于乙酸乙酯和无水乙醇提取物。对石油醚和二氯甲烷提取物作进一步研究,结果显示,各月份石油醚提取物对原头节的杀灭作用由强到弱依次为6>3>11>4>2>5>10>8>12>7>1>9,其中6月份提取物的作用最强,LC50为2.577 μg/ml,95%置信区间为0.85~6.22 μg/ml;各月份二氯甲烷提取物的杀灭作用由强到弱依次为:11>5>10>4>7>12>6>9>8>2>3>1,其中11月份的作用最强,LC50为21.85 μg/ml,95%置信区间为12.38~36.28 μg/ml。 结论 大叶桉桉叶中含有抗细粒棘球蚴的活性成分,值得做进一步的分离、分析和鉴定。

关键词: 大叶桉, 细粒棘球蚴, 原头节, 半数致死浓度

Abstract: Objective To investigate the effect of Eucalyptus robusta leaves extract against Echinococcus granulosus protoscolices in vitro. Methods Mature leaves of Eucalyptus robusta were collected on 24th day in each month from January to December 2012, and air-dried in the room. Ultrasonic extraction of the leaves was done with 4 solvents with different polarity, petroleum ether, dichloromethane, ethyl acetate and anhydrous ethanol. Protoscolices were incubated with the extract at various concentrations for 72 h, and mortality and median lethal dose(LC50) was calculated. Results The extracts were different in characteristics and yield. The petroleum ether extract was in the form of black oil, while dichloromethane, ethyl acetate and anhydrous ethanol extracts were in the form of dark green, pink and white powder respectively. The average yields by petroleum ether, dichloromethane, ethyl acetate, and anhydrous ethanol were 4.4%, 2.1%, 2.3% and 2.3%, respectively. The extract yield was highest for petroleum ether, with a yield of 5.4% in May. The mortality of protoscoleces in all monthly groups of petroleum ether and dichloromethane extracts reached 100% with the concentration of 100 μg/ml and the same mortality reached in most groups of petroleum ether extracts with the concentration of 50 μg/ml. The effects of dichloromethane extracts were less than petroleum ether extracts, but significantly stronger than those of ethyl acetate and ehanol extracts. Further studies conducted on petroleum ether and dichloromethane extracts showed, the lethal effect of petroleum ether extract ranked in month of preparation from strong to weak as June>March>November>April>February>May>October>August>December>July>January>September. In June, the LC50 was 2.577 μg/ml and 95% confidence interval was 0.85-6.22 μg/ml. The lethal effect of dichlorom ethane extract ranked in month of preparation from strong to weak as November>May>October>April>July>December>June>September>August>February>March>January. In November, the LC50 was 21.85 μg/ml, and 95% confidence interval was 12.38-36.28 μg/ml. Conclusion The Eucalyptus robusta leaves contain potential compounds against Echinococcus granulosus. Further experiments of isolation, analysis and identification are needed.

Key words: Eucalyptus robusta, Echinococcus granulosus, Protoscolices, Median lethal dose