The Bushenhuoxue formula improves prothrombotic state in recurrent spontaneous abortion: network pharmacology and experimental validation.

Introduction: Bushenhuoxue formula is a traditional Chinese medicine formula with relatively safe clinical effects, but its mechanism in recurrent spontaneous abortion (RSA) is still unclear. Our present study used Network pharmacology an experimental validation to discuss how Bushenhuoxue formula improves prethrombotic state in RSA.

Materials and methods: The active ingredients of Bushenhuoxue formula (Drug) were acquired from our previous study. The putative targets of ZYP relevant to AS were obtained from TCMSP, Swiss Target Prediction, STITCH, DisGeNET, and Gene Cards databases. Protein-protein interactions (PPI) network, Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were conducted using the Cytoscape software. Furthermore, in vivo experiments were carried out for target validation in BALB/c mice, collecting placental tissue from different groups, the cell apoptosis by TUNEL assay; the pathology by HE staining; relative mRNA expression by qRT-PCR assay; relative protein expression by IHC and WB assay.

Results: Animal experiments, compared with the NC group, the AT-III, PROG, HCG, APC and t-PA concentrations were significantly depressed (P˂0.05, respectively), Apoptosis cell numbers were significantly up-regulated with PI3K/AKT/HIF-1α VEGF significantly depressing (P˂0.001, respectively). With Bushenhuoxue formula supplement, AT-III, PROG, HCG, APC and t-PA concentrations were significantly improved in RSA model mice; and improved pathological changes and apoptosis cell number in placenta tissues (P˂0.05, respectively). However, with LY294002 supplement, the drug treatment effects were disappeared.

Conclusion: Bushenhuoxue formula improves prethrombotic state in RSA via stimulating PI3K/AKT/HIF-1α/VEGF pathway in vivo.