Discussion on the mechanism of action o f dandelion in the treatment of premature ovarian failure based on network pharmacology and in vitro experiments
Objective To explore the mechanism of dandelion in the treatment of premature ovarian failure by network pharmacology technology and in vitro experiments.Methods The active ingredients of dandelion and potential targets for premature ovarian failure were obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analy-sis Platform,STITCH,BATMANIC-TCM,GEO,Traditional Chinese Medicine Information Database,Bindingdb,and DAVID databases.Target protein-protein interaction network and component-target-disease network were constructed,and KEGG enrichment analysis was performed.Ovarian granulosa cells were cultured in different concentrations of H2O2 for dif-ferent times,and the optimal model conditions were selected to construct a premature ovarian cell senescence model(model group).Then the cells were pretreated with different concen-trations of dandelion sterol,and the optimal concentration of subsequent intervention was selected.The cells were divided into blank group,model group,dandelion sterol-high dose,and low dose groups.The cells in the dandelion sterol-high dose,and low dose groups were pretreated with dandelion sterol for 24 h and H2O2 was added toghter with model group.After the intervention,the cell viability was measured by CCK-8 method,the level of reactive oxygen species(ROS)in the cells was detected by DCFH-DA fluorescent probe,and the hypox-ia-inducible factor-1α(HIF-1α)in the cytoplasm and nucleus of the cells was detected by Western blotting.Results A total of 23 active ingredients of dandelion were screened out from the database,345 corresponding target genes,497 target genes related to premature ovarian failure,and 61 common target genes.The pathways involved HIF-1 signaling pathway,proteoglycan interaction,and so on.When the cells were treated with 60 μmol/L H2O2 for 4 h,the cell viability was closest to the IC50.When the concentration of dandelion sterol was 7.5 and 15.0 μmol/L,the cell viability was higher than that of the model group(P<0.05).The relative fluorescence intensity of ROS in model group was higher than that in blank group,and those in dandelion sterol high-dose and low-dose groups were lower than that in model group(P<0.05).There was no sig-nificant difference in the expression of HIF-1α protein in cytoplasm among the four groups(P>0.05).The expression of nuclear HIF-1α protein in the model group was higher than that in the blank group,and those in dandelion sterol high-dose and low-dose groups were lower than that in model group(P<0.05).Conclusion Dandelion can treat premature ovarian failure through multi-component,multi-target,and multi-pathway,and its representative component dandelion sterol may play a role in the treatment of premature ovarian failure through antioxidant effects.