关键词: 华支睾吸虫, 体外培养, 吡喹酮, 三苯双脒, 左旋咪唑, 青蒿琥酯, 蒿甲醚, 阿苯达唑, 甲苯达唑

Abstract:  Objective   To observe the in vitro effect of praziquantel, tribendimidine, levamisole, artemether, artesunate, albendazole and mebendazole against adult Clonorchis sinensis.  Methods   Seventy rats infected with 50-100 C. sinensis metacercariae for 5-7 weeks were euthanized, and adult C. sinensis were collected from the common bile duct. Three to four worms were placed in each well of a 24-well falcon plate, and treated by Hanks’ balanced salt solution-20% calf serum containing aforementioned drugs at various concentrations. The motor activity and morphology change of the worms were observed under an inverted microscope at 4，24，48 and 72 h post treatment.  Results   Praziquantel could reduce the motor activity of the worms rapidly which resulted in detachment of oral sucker from the well wall, curl of the worm body and emergence of vacuoles from the tegument. The minimal concentration of praziquanel to kill adult C. sinensis was 0.1 g/ml. After adult C. sinensis exposed to tribendimidine at concentrations of 0.5，1 and 10 g/ml, they revealed in paralysis, looseness and stretch of the worm body rapidly or immediately. The minimal concentration of tribendimidine to kill adult worms was 0.05 g/ml. When worms exposed to levamisole at 10 and 20 g/ml, there was a gradual decrease in the worm’s motor activity accompanied by looseness of the worm body. But 48 h post exposure, most worms showed apparently recovery of motor activity. In a higher levamisole concentration of 50 g/ml, all worms revealed in stretch and paralysis which was similar to that induced by tribendimidine. When adult C. sinensis were exposed to artemether or artesunate 10 and 50 g/ml, the motor activity of worm body and oral sucker reduced which accompanied by worm contraction, then followed by loosness of the worm body and emergence of vacuoles along the tegument. At 72 h post exposure, the worm mortalities induced by the two concentrations of the two drugs were about half, respectively. In adult C. sinensis exposed to albendazole and mebendazole at concentrations of 10 and 50 g/ml, only stimulation of motor activity of oral sucker was seen which revealed in vigorous contraction within 24 h post exposure. During 72 h observation period, no any other changes in worm activity and morphology were seen.  Conclusion   Praziquantel and tribendimidine exhibit strong in vitro killing effect on adult C. sinensis. The minimal concentration of levamisole used to kill adult worm is 50 times higher than that of tribendimidine. The higher concentrations of artemether and artesunate show slower action to reduce the worm activity and kill part of the worms. Higher concentrations of albendazole and mebendazole exhibit no killing effect on C. sinensis, besides stimulating the motor activity of worm oral sucker.

Key words: Clonorchis sinensi, In vitro culture, Praziquantel, Tribendimidine, Levamisole, Artemether, Artesunate, Albendazole, Mebendazole