Study On Killing Effect Of Artesunate On Protocercariae Of Echinococcus Multilocularis In Vitro

Objective:The purpose of this experiment is to observe the killing effect of artesunate on the protoscolex of echinococcus multilocularis in vitro,and to explore the therapeutic value of artesunate on alveolar echinococcosis(AE).Methods:Artesunate was dissolved in NaHCO? solution and mixed with RPMI-1640 medium at different concentrations(250 ?g/mL,200 ?g/mL,150 ?g/mL,100 ?g/mL,50 ?g/mL).This is the experimental group.The control groups are 250 ug/mL albendazole,250 ?g/mL praziquantel and blank control.Firstly,the protoscolex of echinococcus multilocularis was cultured with drugs in vitro.We observe the activity and mortality of protoscolex under light microscopy after 1% eosin staining within 48 hours.Then we observe the changes of protoscolex dynamically by confocal microscopy.The structure of protoscolex was observed by scanning electron microscopy(SEM)after death.Finally,we use statistical methods to analyze the data.Result:1.After 28 hours of drug intervention,the mortality rate of artesunate 250 ?g/mL group(99.96±0.03%)was significantly higher than that of 250 ?g/mL albendazole,250 ?g/mL praziquantel and blank control group(9.21±0.16%,10.00±0.24%,3.68±0.07%)(P<0.05).After 48 hours of drug intervention,the mortality rates of artesunate group(200 ?g/mL,150 ?g/mL,100 ?g/mL,50 ?g/mL)were 99.33±0.54%,91.31±1.09%,77.05±0.70% and 47.77±1.46% respectively,which were higher than those of 250 ug/mL albendazole group,250 ?g/mL praziquantel group and blank control group(20.00±1.77%,20.07±1.46% and 7.21±0.13%).There was statistical significance(P<0.05).The mortality rate of protocephalus after was proportional to the intervening time and drug concentration.2.Then we analyzed the morphology of multilocular protocercariae after artesunate intervention under optical microscope,scanning electron microscope(SEM)and laser confocal microscope.It was found that the surface structure and internal structure of multilocular protocercariae changed greatly after artesunate intervention.The bodysurface shrank,the internal particles disappeared,the top broke through and the activity of multilocular protocercariae decreased until death.Conclusion:Artesunate has obvious killing effect on the protoscolex of echinococcus multilocularis in vitro.