Tang Zi-Hao, Liu Ke-Yue, Mei Jun, Gao Xing-Zheng
Department of Pathogenic Biology, Jiujiang University Medical College, Jiujiang 332000, China.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2010 Dec 30;28(6):416-21.
To observe the effect of a mixture of dihydroartemisinin and metronidazole on ultrastructure of Trichomonas vaginalis trophozoites in vitro for exploring trichomonacidal mechanism of the drug mixture.
The trophozoites were cultivated with liver extract solution medium that contained 2.5 x 10(6) parasites/ml. There were dihydroartemisinin 0.5 mg/ml and metronidazole 0.002 mg/ml in the experimental tubes of the drug mixture group. Groups of control (without drug), dihydroartemisinin (1 mg/ml) and metronidazole (5 mg/ml) were established and performed in the same experimental conditions. The parasites were observed by scanning and transmission electron microscopes after having treated with the drugs at 37 degrees for 3.5-5 h.
Under scanning electron microscope, the cell membrane of T. vaginalis treated only with dihydroartemisinin for 35 h was damaged, part of pellicle peeled off. Although the surface of the trophozoites treated only with metronidazole for 5 h showed many small bubbles and hollows, the cell membrane looked integral. However, surface of the parasite exposed to the drug mixture for 3.5-4.2 h showed deep folds and cracks, the cell membrane was damaged and even peeled off. When the cell ruptured, the nucleus, axostyle, pelta and hydrogenosomes were exposed, and the cytoplasm spilled out. Transmission electron microscopy showed that the membrane system of the trophozoites treated only with dihydroartemisinin for 3.5 h was damaged considerably. The cytoplasm of damaged parasite spilled out. The cytoplasm of the parasite treated only with metronidazole for 3.5-5 h was damaged seriously. Vacuoles and crevices were visible in the cytoplasm. The cell membrane and the content of the parasites treated with the drug mixture for 3.5-4.5 h were damaged seriously. There were some vacuoles and crevices, dilated endoplasmic reticulum, injured and deformed hydrogenosomes in the cytoplasm. The cell organelles mostly disappeared. Crevices also existed in the nucleus. The nuclear membrane fractured and even disappeared.
The acting targets of dihydroartemisinin and metronidazole to T. vaginalis trophozoite were different, and a combination of the two drugs shows stronger effect in killing the parasites.
观察双氢青蒿素与甲硝唑合剂对体外阴道毛滴虫滋养体超微结构的影响,以探讨该合剂的杀滴虫机制。
用含2.5×10⁶个寄生虫/ml的肝浸液培养基培养滋养体。药物合剂组实验管中含双氢青蒿素0.5mg/ml和甲硝唑0.002mg/ml。设立对照组(无药物)、双氢青蒿素组(1mg/ml)和甲硝唑组(5mg/ml),在相同实验条件下进行。将药物处理后的寄生虫于37℃作用3.5 - 5小时后,用扫描电子显微镜和透射电子显微镜观察。
扫描电子显微镜下,仅用双氢青蒿素处理35小时的阴道毛滴虫细胞膜受损,部分表膜脱落。仅用甲硝唑处理5小时的滋养体表面有许多小气泡和凹陷,但细胞膜看起来完整。然而,暴露于药物合剂3.5 - 4.2小时的寄生虫表面出现深褶皱和裂缝,细胞膜受损甚至脱落。当细胞破裂时,细胞核、轴柱、盾片和氢化酶体暴露,细胞质溢出。透射电子显微镜显示,仅用双氢青蒿素处理3.5小时的滋养体膜系统受损严重。受损寄生虫的细胞质溢出。仅用甲硝唑处理3.5 - 5小时的寄生虫细胞质受损严重。细胞质中可见空泡和缝隙。用药物合剂处理3.5 - 4.5小时的寄生虫细胞膜和内容物受损严重。细胞质中有一些空泡和缝隙,内质网扩张,氢化酶体受损变形。细胞器大多消失。细胞核中也有缝隙。核膜破裂甚至消失。
双氢青蒿素和甲硝唑对阴道毛滴虫滋养体的作用靶点不同,两药联合对杀灭寄生虫显示出更强的效果。