The Expression Of Antioxidant Enzymes Was Regulated By The MAPK-Nrf2 Signaling Pathway In E.granulosus Protoscoleces

Objective:Current research shows that mitogen-activated protein kinases?MAPKs?could increase the phosphorylation levels of nuclear factor erythroid-2-related factor 2?Nrf2?and induce the transcription of downstream target genes.However,very little is known about the relationship between MAPKs and Nrf2 in Echinococcus granulosus protoscoleces.The present work sought to explore the relationship between MAPKs and Nrf2 signaling pathways in E.granulosus protoscoleces.Methods:Primarily.Echinococcus granulosus protoscoleces were collected aseptically from cysts of infected sheep and In vitro cultured with RPMI 1640.The experiment was divided into the following groups:The first group,protoscoleces of E.granulosus were pretreated with various concentrations?0-1mM?of propofol for 24h.The second group,protoscoleces of E.granulosus were pretreated with 0.5 mM of H₂O₂ for 6h.The third group,protoscoleces of E.granulosus were pretreated with various concentrations?0-1mM?of propofol for 8h followed by exposure to 0.5 mM of H₂O_2.The fourth group,PD98059?an inhibitor of the ERK?,SB202190?an inhibitor of p38 MAPkinase?,and SP600125?an inhibitor of JNK?were used in pretreatment of protoscoleces for 2h,and then protoscoleces were co-treated with1mM propofol for another 8h,protoscoleces were incubated for 6h in the presence of H₂O₂Cultures were followed microscopically to determine the viability assays of protoscoleces.Drawing dynamic curve after the results are representative of 3 independent experiments.The activity of HO-1 and ROS between different group were detected using an assay kit.The expression levels of Nrf2 and HO-1 between different group elucidated through western blotting.Results:The relative survival rate of protoscoleces treated with 0.25 mM,0.5 mM,0.75mM,1 mM propofol was over 95%.The survival rates of protoscoleces treated with 0.5mM H₂O₂ for 6h were 38%.Protoscoleces were treated with propofol?0.5-1 mM?for 8 h,after which,the protoscoleces were exposed to 0.5 mM H₂O₂ in fresh medium for 6 h.The survival rates of protoscoleces treated with 0.5 mM,0.75 mM,and 1 mM propofol were 48%,58%,and 65.5%,respectively.According to our previously described methods,protoscoleces were pretreated with PD98059,SB202190,SP600125 for 2 h,co-treated with 1 mM propofol for an additional 8 h,and then exposed to 0.5 mM H2O2 in fresh medium for 6 h.The survival rates of protoscoleces were 61%,36%,and 34%.Compared with the control group,protoscoleces mortality increased significantly?P<0.05?.The protoscoleces in each treatment group were treated with DCFH-DA probe for 1h.Results shows that treatment with propofol significantly reduced H₂O₂-induced ROS generation,but co-incubation of the protoscoleces with H₂O₂,and MAPK inhibitors increased protoscolece ROS productionP<0.05).Using a colorimetric HO-1 assay kit,it was found that propofol and H2O2 increased the expression HO-1 compared with that of the control group.JNK or p38MAPK signaling pathways were Significantly inhibited propofol-induced HO-1 expression.Propofol increased the expression of Nrf2 and HO-1 compared with that of the control group.Co-incubation of the protoscoleces with propofol,H2O2,and SP600125 or SB202190 reduced the expression of Nrf2 and HO-1,PD98059 reduced the expression of Nrf2 and HO-1 slightly.Result detected by western blotting.Conclusions:1.H₂O₂ and inhibitor of MAPK induces apoptosis of protoscoleces and induces ROS generation in E.granulosus protoscoleces.2.Inhibitor of MAPK reduces in the expression of Nrf2 and HO-1 induced by H2O2 and propofol.