基于网络药理学、分子对接技术和动物实验验证探讨阳和消囊汤治疗多囊卵巢综合征的作用机制OA

Objective To investigate the mechanisms of Yanghe Xiaonang Decoction in treating polycystic ovary syndrome(PCOS)using network pharmacology,molecular docking techniques,and animal experiments.Methods The chemical constituents of Yanghe Xiaonang Decoction and their potential targets for treating PCOS were screened using The Encyclopedia of Traditional Chinese Medicine(ETCM)v2.0,Traditional Chinese Medicine Database and Analysis Platform(TCMIP)v2.0,GeneCard,DisGeNET,Malacards,and the National Center for Biotechnology Information(NCBI)Gene Database,followed by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses.The binding interactions between core targets and active components were validated by molecular docking techniques.In animal experiments,72 female rats were divided into six groups:a control group,a model group,low-,medium-,and high-dose Yanghe Xiaonang Decoction groups,and a clomiphene citrate group by the random number table method,with 12 animals in each group.Except for the control group,all the other groups were administered letrozole via gavage for 21 consecutive days to establish a PCOS rat model.The following parameters were determined in each group:body weight and ovarian histopathological changes,follicle-stimulating hormone(FSH),luteinizing hormone(LH),testosterone(T),estradiol(E2),and adiponectin(ADPN)levels,along with phosphoinositide 3-kinase(PI3K),protein kinase B1(AKT1),forkhead box O1(FOXO1),sirtuin 1(SIRT1),and small mother against decapentaplegic 3(SMAD3)mRNA and protein expression levels.Results Network pharmacology sieved out 304 targets through which Yanghe Xiaonang Decoction acted on PCOS.Topological analysis obtained 74 key network targets,among which 30 core targets were selected using the Maximal Clique Centrality(MCC)algorithm.GO functional enrichment analysis revealed 452 cellular biological processes,61 cellular components,and 108 molecular functions.KEGG pathway enrichment analysis showed that the primary pathways by which Yanghe Xiaonang Decoction treats PCOS included the thyroid signaling pathway,growth hormone synthesis/secretion/interaction pathway,FoxO signaling pathway,and PI3K/Akt signaling pathway.Molecular docking analysis extracted 124"active ingredient-target"pairs whose binding free energy demonstrated stable binding affinities.According to animal experiments,compared to the model group,the low-,medium-,and high-dose Yanghe Xiaonang Decoction groups,alongside the clomiphene citrate group exhibited significantly reduced serum LH and T levels,as well as significantly elevated FSH,E2,and ADPN levels(P＜0.05).The low-dose Yanghe Xiaonang Decoction group showed increased SIRT1 mRNA expression along with decreased FOXO1 and SMAD3 mRNA expression(P＜0.05).The medium-and high-dose Yanghe Xiaonang Decoction groups,alongside the clomiphene citrate group demonstrated increased PI3K,AKT,and SIRT1 mRNA expression along with decreased FOXO1 and SMAD3 mRNA expression(P＜0.05).Conclusion Yanghe Xiaonang Decoction inhibits apoptosis in granulosa cells to promote follicular development and ovulation function in PCOS model rats by regulating downstream genes and their expressions in the FOXO1 pathway via activating the PI3K/AKT signaling pathway in ovarian tissues.