Mechanism of Dehydrocamptothecin in the Treatment of Echinococcosis Granulosa
Objective To predict and analyze the mechanism of dehydrocamptothecin(HM)in the treatment of echinococcosis granulosa(EG)based on the network pharmacology method.Methods HM-related genes were obtained from the Traditional Chinese Medicine System Pharmacology Database and Analysis Platform(TCMSP),SwissTargetPrediction,and Drugbank databases.The target genes of EG were obtained from GeneCards,DISGENET database,and Drugbank databases.The predicted targets of HM and genes related to EG were mapped to obtain the target genes of HM acting on EG.The network topology attribute analysis was performed by importing them into the String database to screening important target proteins to construct protein-protein interaction(PPI)network and component-disease-target network.DAVID database was used for gene ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis to screen gene targets related to DNA damage.Autodock Vina and Pymol softwares were used to validate and display molecular docking of relevant targets.Echinococcus granulosus with activity not less than 98%was randomly divided into the blank control group,1%DMSO group,25 µmol/L HM group,50 µmol/L HM group,and 100 µmol/L HM group through in vitro experiments.Quantitative reverse transcription-polymerase chain reaction(qRT-PCR)method and Western blot method were selected to verify the intervention effect of HM on the expression levels of TP53,TopoⅠ,H2AX,and ATM mRNA and proteins.Results A total of 105 targets of HM and 88 genes related to EG were obtained,with two intersecting targets.The main targets of HM acting on EG were MAPK3,MAPK1,TP53,HSPAA1,MAPK14,Jun,and the mechanism might involve MAPK signaling pathway,ERBB signaling pathway,DNA damage pathway and DNA repair pathway,cell apoptosis,and other processes.In vitro validation method showed that compared with those in the blank control group,the expression levels of TP53 and TopoⅠ mRNA and proteins showed a downward trend,while the expression levels of H2AX and ATM mRNA and proteins showed an upward trend in the 25 µmol/L HM group,50 µmol/L HM group,and 100 µmol/L HM group(P<0.05).Conclusion HM can treat EG through various biological processes and related signaling pathways.HM is involved in the regulation of DNA damage pathway and DNA repair signaling pathway in the treatment of EG,and TP53,TopoⅠ,H2AX,and ATM may be the targets.This study lays a foundation for further elucidating the molecular mechanism of HM in the treatment of EG.
dehydrocamptothecinechinococcosis granulosaDNA damage and DNA repairqRT-PCRnetwork pharmacologymechanism
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