RESUMO
Objective To investigate the underlying mechanism of Huayu Xiaozheng granule(HYXZ)in the treatment of ectopic pregnancy(EP).Methods The potential ingredients and targets of HYXZ were identified and screened.PPI network,GO enrichment and KEGG pathway analysis were established to understand the potential and intricate mechanism of HYXZ in treating EP.The effects of HYXZ on HTR-8/SVneo cells migration and invasion were detected by wounding healing assay and transwell migration assay respectively,as well as the influence of HYXZ granule on JAK/STAT pathway and associated proteins were assessed by western blot under the control of JAK/STAT pathway inhibitor and agonist.Results A total of 375 active ingredients,45 known targets and 207 predicted targets of HYXZ were obtained.A PPI network has constructed to know the relationship of 252 common targets of drug targets and disease targets,from which we screened out 15 key targets:STAT3,MMP9,TLR4,HIF1A and others.Scratch test showed that HYXZ inhibited the migration of the HTR-8/SVneo cells cells in a time and dose-dependent manner;Transwell test showed that the number of transmembrane cells decreased significantly(P<0.05)after 48 h with various concentrations of HYXZ;Western blot experiment showed that the expression of JAK/STAT pathway protein decreased significantly(P<0.05);and the expression of its related proteins uPA,HIF1and N-cadherin were inhibited(P<0.05),the tendency of which was consistent with the inhibitor AG490,but opposite to the agonist IL-6.Conclusion HYXZ inhibited JAK/STAT pathway,and associated proteins uPA,HIF1A and N-cadherin,thereby suppressing the mobility and invasiveness of the HTR-8/SVneo cells,which is conductive to clarify the mechanism of HYXZ in the treatment of early EP.