摘要
This study was to investigate the protective effect of paeoniflorin(PF)on hydrogen peroxide-induced injury.Firstly,“SMILES”of PF was searched in Pubchem and further was used for reverse molecular docking in Swiss Target Prediction database to obtain potential targets.Injury-related molecules were obtained from GeenCards database,and the predicted targets of PF for injury treatment were selected by Wayne diagram.For mechanism analysis,the protein-protein interactions were constructed by String,and the KEGG analysis was conducted in Webgestalt.Then,cell viability and cytotoxicity assay were established by CCK8 assay.Also,the experimental cells were allocated to control,model(200μmol·L^(−1) H_(2)O_(2)),SB20358010μmol·L^(−1)(200μmol·L^(−1) H_(2)O_(2)+SB20358010μmol·L^(−1)),PF 50μmol·L^(−1)(200μmol·L^(−1) H_(2)O_(2)+PF 50μmol·L^(−1)),and PF 100μmol·L^(−1)(200μmol·L^(−1) H_(2)O_(2)+PF 100μmol·L^(−1))groups.We measured the intracellular ROS,Hoechst 33258 staining,cell apoptosis,the levels of Bcl-xl,Bcl-2,Caspase-3,Cleaved-caspase3,Cleaved-caspase7,TRPA1,TRPV1,and the phosphorylation expression of p38MAPK.There are 96 potential targets that may be associated with PF for injury treatment.Then,we chose the“Inflammatory mediator regulation of TRP channels”pathway for the experimental verification from the first 10 KEGG pathway.In experimental verification,H_(2)O_(2) decreased the cell viability moderately(P<0.05),and 100μmol·L^(−1) PF increased the cell viability significantly(P<0.05).Depending on the difference of intracellular ROS fluorescence intensity,PF inhibited H_(2)O_(2)-induced reactive oxygen species production in Schwann cells.In Hoechst 33258 staining,PF reversed the condensed chromatin and apoptotic nuclei following H_(2)O_(2) treatment.Moreover,Flow cytometry results showed that PF could substantially inhibit H_(2)O_(2) induced apoptosis(P<0.05).Pretreatment with PF obviously reduced the levels of Caspase3,Cleaved-caspase3,Cleaved-caspase7,TRPA1,TRPV1,and the
基金
the National Natural Science Foundation of China(No.81874404).
