Arsenic Trioxide Induced Apoptosis Of Echinococcus Granulosus Protoscoleces Through Endoplasmic Reticulum Stress

Objective:Our preliminary results showed that arsenic trioxide?As₂O₃?can inhibit the growth ofprotoscoleces of Echinococcus granulosus,destroyed the original structure of protoscoleces,activated of the apoptosis pathway mediated by Caspase-3,However,other possible pathways of apoptosis have not been studied.It has been studied that As₂O₃ induces tumor cell apoptosis through endoplasmic reticulum stress and oxidative stress,.This study to explore the possible mechanism of he induced apoptosis of protoscoleces of Echinococcus granulosus.Methods:In this study,we use different concentration of As₂O₃ for different time in protoscoleces of Echinococcus granulosus,with fresh sheep liver disease from infection of Echinococcus granulosus protoscoleces as research object.Protoscoleces activity after treatment of As₂O₃ was detected by 0.1%eosin,ROS in protoscoleces was detected by Fluorescence microscopy,at the same time,we detected the related antioxidant enzymes GSH,SOD activity,the expression of Caspase-3 enzyme viability in protoscoleces by kit,flow cytometry was used to detect the level of Ca²⁺in protoscoleces,the expression level of caspase-12?GRP78?Bax and Bcl-2 was detected by Western-blot analysis.Results:The staining results:the activity of protoscoleces decreased gradually with increasing time and concentration of As₂O₃;Kit results:As₂O₃ can increase the activity of caspase-3 in protoscoleces?P<0.05?,increased ROS activity,and decreased SOD,GSH activity?P<0.05?;Flow cytometry results:compared with the control group,the concentration of Ca²⁺in drug groups were significantly higher,and has a time and concentration dependence?P<0.05?.The results of western-blot analysis:the expression level of GRP78?caspase-12 and Bax increased significantly,expression level of Bcl-2 protein was decreased,and showed a concentration dependence.Conclusion:As₂O₃ induced apoptosis in protoscoleces of Echinococcus granulosus by endoplasmic reticulum stress pathway and oxidative stress pathway,provides a new drug targets for hepatic hydatid disease.