基于网络药理学及实验验证探讨养血化瘀汤对卵巢过度刺激综合征的作用机制OA

Objective To analyze the potential mechanism of Yangxue Huayu Decoction(YXHYD)in the treatment of Ovarian Hyperstimulation Syndrome(OHSS)based on a combination of the network pharmacology and experimental validation.Methods The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)was employed as the primary source of data to screen active compounds of YXHYD.To ensure the comprehensiveness and reliability of target information,the predicted targets were integrated with data from authoritative databases such as DrugBank.Potential targets were retrieved by MOL.ID and were subsequently annotated with standardized information using the UniProt database.An"active compound-target"network was constructed with software Cytoscape 3.8.2 using the relevant files and analyzed by network topology.Potential therapeutic targets of YXHYD in the treatment for OHSS were identified by integrating drug-disease targets from the GeneCards and OMIM databases.A protein-protein interaction network for these potential therapeutic targets was constructed using the STRING platform.Gene Ontology(GO)functional annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed using R software.Molecular docking experiments were conducted to predict the result.Sixty rats were divided into six groups(10 for each group)by a random number table:control group,model group,positive group,low-,mid-and high-dose YXHYD groups.With the exception of the control group,an OHSS rat model was established in all other groups.After successful modeling,the positive group was administered with 25 g/(kg·d)aspirin;low-,mid-and high-dose YXHYD groups were gavaged with YXHYD 2.27,4.55 and 9 g/(kg·d)respectively;control group and model group received an equivalent volume of physiological saline.The intervention lasted for one week.The ovarian weight was compared.The expression level of key targets were detected by quantitative real-time polymerase chain reaction(RT-qPCR)and Western blot.Results According to network pharmacological analysis,YXHYD may alleviate syndrome of OHSS by regulating the estrogen signaling pathway,VEGF signaling pathway and PI3K-Akt signaling pathway.In animal experiments,rats in all YXHYD groups significantly reduced ovarian weights compared with those in the model group(P<0.05,P<0.01).Furthermore,molecular level detection results demonstrated that VEGF,Akt,and ESR mRNA expression in ovarian tissue significantly decreased in the mid-and low-dose YXHYD groups(P<0.01,P<0.001).Meanwhile,Western blot revealed a significant decrease in the protein expression levels of Akt,ESR,and VEGFA in the YXHYD-treated groups(P<0.001).Conclusion YXHYD might exert its preventive and therapeutic effects on OHSS by inhibiting VEGF-mediated PI3K/Akt signaling activation,down-regulating ESR expression,and blocking the VEGF-ESR positive feedback loop,thereby alleviating the hyperpermeability of vascular permeability in OHSS.