使君子提取物体外抗蓝氏贾第鞭毛虫效果的研究

曹晓琴; 童杨; 方振峰; 施璐; 房辉; 刘骥才

doi:10.13881/j.cnki.hljxmsy.2022.07.0101

使君子提取物体外抗蓝氏贾第鞭毛虫效果的研究

江汉大学医学院

详细信息

作者简介:
曹晓琴(1982—),女,副教授,博士,研究方向为药物分析及食品安全检测,caoxiaoqin012@jhun.edu.cn

通讯作者:
刘骥才(1989—),男,讲师,硕士,研究方向为中药活性成分,315379839@qq.com

中图分类号: S853.74
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出版历程
- 收稿日期: 2022-07-11
- 刊出日期: 2023-05-09

Study on Combretum indicum extracts against Giardia intestinalis in vitro

College of Medicine, Jianghan University

摘要

摘要: 为了观察中药使君子提取物体外抗蓝氏贾第鞭毛虫（简称贾第虫）的作用效果，试验分别采用乙醇和去离子水对使君子进行提取，得到使君子醇提物和使君子水提物，设置使君子醇提物组、使君子水提物组、甲硝唑对照组（3组各分别设置0.312 5,0.625,1.25,2.5,5 mg/mL5个药物浓度）及空白对照组（TYI-S-33培养基）,分别在37℃厌氧培养滋养体浓度为8.0×10～5～4.2×10～6个/mL的贾第虫24 h,收集贴壁虫体，计算半数抑制浓度(IC₅₀);用IC₅₀的使君子醇提物、使君子水提物和甲硝唑分别作用于贾第虫，依次培养0,2,4,6,8,12,24 h,同时设空白对照，计算出药物达IC₅₀的最短作用时间；对作用时间分别为6 h和12 h的使君子醇提物组和空白对照组进行倒置光学显微镜、扫描电镜和透射电镜下观察，分析使君子醇提物对贾第虫滋养体超微形态结构的影响。结果表明：三种药液对贾第虫均有一定的抑制作用。使君子醇提物、使君子水提物和甲硝唑的IC₅₀分别为1.060,8.145,0.325 mg/mL,达IC₅₀的最短时间分别为5.28 h、5.95 h和3.85 h。IC₅₀浓度下，培养6 h及以上时，使君子醇提物组与甲硝唑对照组虫体浓度差异不显著（P>0.05）,而使君子水提物组虫体浓度均显著高于甲硝唑对照组（P<0.05）。使君子醇提物作用6 h后滋养体虫体变形，鞭毛受损；作用12 h后引起细胞膜破裂，细胞内容物外溢，吸盘微管部分结构崩解，滋养体开始死亡。说明使君子具有抑制贾第虫生长的作用，且醇提物比水提物抑制效果更强，呈浓度依懒性。IC₅₀浓度的使君子醇提物与甲硝唑对虫体抑制效果相当。
- 蓝氏贾第鞭毛虫 /
- 使君子醇提物 /
- 使君子水提物 /
- 生长抑制 /
- 形态结构
Abstract: In order to observe the effects of traditional Chinese medicine Combretum indicum extracts against Giardia intestinalis（referred to as Giardia） in vitro, in this experiment, ethanol and deionized water were used to extract Combretum indicum to obtain Combretum indicum alcohol extracts and Combretum indicum water extracts.Combretum indicum alcohol extracts group, Combretum indicum water extracts group, and metronidazole control group（the concentrations were 0.312 5, 0.625, 1.25, 2.5, 5 mg/mL）, and blank control group（TYI-S-33 medium）, were set respectively.Giardia was cultured anaerobically at 37 ℃ at a concentration of 8.0×10~5-4.2×10~6 trophozoites/mL for 24 h, respectively. Adherent Giardia were collected and half of the inhibitory concentration(IC₅₀) was calculated. The Giardia were exposed to Combretum indicum water extracts, Combretum indicum alcohol extracts, and metronidazole using IC₅₀, respectively; and at the same time a blank control was set to culture for 0, 2, 4, 6, 8, 12, 24 h. The shortest action time of the drugs up to IC₅₀ was calculated. The IC₅₀ group of Combretum indicum alcohol extracts and blank control group with action time of 6 h and 12 h respectively were observed under inverted light microscopy, scanning electron microscopy and transmission electron microscopy to analyze the effects of Combretum indicum alcohol extracts on the ultramorphological structure of Giardia trophozoites. The results showed that the three medical solutions had a certain inhibitory effect onGiardia. The IC₅₀ of Combretum indicum alcohol extracts was 1.06 mg/mL; the IC₅₀ of Combretum indicum water extracts was 8.145 mg/mL, and the IC₅₀ of metronidazole was 0.325 mg/mL; and the shortest time to IC₅₀ was 5.28 h, 5.95 h and 3.85 h, respectively. At IC₅₀ concentration, when incubated for 6 h or more, there was no significant difference in the Giardia concentration between the Combretum indicum alcohol extracts group and the metronidazole control group（P＞0.05）. During the entire culture period, the Giardia concentration in Combretum indicum water extracts group was significantly higher than that in metronidazole control group（P＜0.05）. After 6 h of action of Combretum indicum alcohol extracts, the trophozoite body was deformed, and the flagella were damaged. After 12 h of action, the cell membrane ruptured; the cell contents overflowed; part of the structure of the suction cup microtubules disintegrated, and trophozoites began to die. The results suggested that Combretum indicum had the effect of inhibiting the growth of Giardia, and the alcohol extracts had a stronger inhibitory effect than water extracts, which was concentration dependence. The IC₅₀ concentration of Combretum indicum alcohol extracts was comparable to metronidazole on inhibition effect of Giardia.
- Giardia intestinalis /
- Combretum indicum alcohol extract /
- Combretum indicum water extract /
- growth inhibition /
- morphological structure

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