Study On Lycorine And Lycorine Combined With Albendazole Against Echinococcus Granulosus Protoscoleces In Vitro

Objective: To explore the anti-Echinococcus Granulosus protoscoleces(EgPSC)of lycorine by comparing the efficacy of lycorine,lycorine combined with albendazole and albendazole in vitro.And the mechanism of resistance of lycorine to Eg PSC was further investigated.Methods: The protocolizes were collected from Liver hydatid cysts of sheep and cultured for three days in vitro,then treated with lycorine(100?mol/L,250?mol/L,500?mol/L,750?mol/L,1000?mol/L),lycorine combined with albendazole(100?mol/L+50?mol/L)and albendazole(50?mol/L)respectively.By eosin staining,the viability of Eg PSC was recorded at 1th day,3th day,5th day,7th day,9th day.And the statistical analyses of the viability of protoscolices were performed using Two-way Repeated Measures ANOVA.Then to measure the indexes of apoptosis(Bcl-2?Bax?caspase-9 and caspase-3)and mitochondrial injury(mitochondrial membrane potential)by western blot(WB)and Immunofluorescent in 7th day.Results: 1.The results showed that after 9 days of exposure to 100?mol/L,250?mol/L,500?mol/L,750?mol/L and 1000?mol/L of lycorine,the viability rate of protoscolices were 72.70%,36.65%,14.09%,1.88% and 0%.Between the lycorine and control groups,there were significantly different(P<0.05);Among different concentration groups,the difference was statistically significant(P<0.05);At different time points,the difference were statistically significant in the lycorine treatment groups(P<0.05).2.The results showed that after 9 days of exposure to lycorine combined with albendazole,albendazole and lycorine,the viability rate of protoscolices were 50.20%?90.22% and 72.70%.Between the Combined group and Lycorine group,Albendazole group there were significantly different(P<0.05).3.After 7 days,between the control groups and DMSO treated group,there were no different in the level of Bcl-2,Bax,caspase-9 and caspase-3(P > 0.05).Among the lycorine-treated subgroups,the results suggesting positive correlation between the levels of Bax,caspease-3,caspease-9 with the lycorine concentrations(P<0.05),and the level of Bcl-2 was negatively correlated with lycopene concentration(P<0.05).4.An intense green fluorescence was observed in lycorinetreated groups but less visible in controls groups,and the intensity of green light increases with the dose of lycorine among lycorine-treated subgroups.Conclusion: 1.results indicated the lycorine was efficacious against Eg PSC in virto,and has dose-dependent effect and time-dependent.2.The lycorine and albendazole have synergistic action against Eg PSC in vitro.3.Lycorine can induce the apoptosis of Echinococcus granuloses protocolizes via mitochondrial pathways.