Effect Of AZD2014 On Protoscoleces Of Echinococcus Granulosus In Vitro

Objective: Since AZD2014 is an inhibitor of TOR protein kinase,we have observed the activity of Echinococcus granulosus which cultured by different dose of AZD2014 in vitro,and then obtained the relative optimal concentration of AZD2014 acting on Echinococcus granulosus and that the TOR protein kinase exist in the Echinococcus granulosus.AZD2014 could block the PI3K-AKT-TOR of cell signaling pathway by blocking the TOR protein kinase activity in the granulosus,and thereby kill the granulosus in the vesicle.As a targeting drug,AZD2014 provides a theoretical and research basis in the treatment of echinococcosis.Methods: 1.Echinococcus granulosus were cultured by different concentrations of AZD2014 in vitro.The survival rate was determined by using 0.1% eosin staining and an optical microscope for 15 consecutive days.As well as the survival rate was counted by us.2.Echinococcus granulosus were cultured by different concentrations of AZD2014 in vitro for 4 days.And then we took the protein supernatant from those granulosus,and used Western-blot to verify the expression of protein factors which belong to TOR downstream,and used ELISI kit to detect the expression of Caspase-3 enzyme activity;3.We used 100μM AZD2014 to culture Echinococcus granulosus for 1 day,4 days,and 7 days.And then we took the protein supernatant from those granulosus,and used Western-blot to verify the expression of protein factors which belong to TOR downstream,and used ELISI kit to detect the expression of Caspase-3 enzyme activity.Results: 1.With the increase of culture concentration of AZD2014,the survival rate of Echinococcus granulosus gradually decreased.With the extension of culture time of AZD2014,the survival rate of Echinococcus granulosus decreased gradually.2.Western-blot validated the downstream protein factor of TOR protein kinase.The expression of AKT,p-S6K1 and p-4EBP1 decreased gradually with the increase of AZD2014 concentration,and decreased gradually with the prolongation of AZD2014 culture time(P<0.05).3.The activity of Caspase-3 which verified by ELISI kit increased gradually with the increase of AZD2014 concentration,and the prolongation of AZD2014 culture time(P<0.05).Conclusion: 1.AZD2014 has the effect of killing Echinococcus granulosus by inhibiting the TOR protein kinase in granulosus;2.AZD2014 has the time and dose dependence in killing Echinococcus granulosus which cultured in vitro.