Introduction: This study aims to assess the therapeutic mechanisms of Zishen Yutai Pills (ZYP) in treating recurrent spontaneous abortion (RSA) using network pharmacology analysis and in vitro validation of the molecular pathways relating to angiogenesis. Methods: An online database screening-based strategy was applied for chemical constituent and target screening-based strategy was applied for chemical constituent and target collection, as well as target prediction. Network pharmacological analysis was conducted using protein-protein interaction data. Molecular docking was further employed to validate the active components of ZYP and the central target proteins. The tube formation assay was conducted in human umbilical vein endothelial cells (HUVECs) with the intervention of active constituents in ZYP. The expression of vascular endothelial growth factor A (VEGFA) and protein kinase B (AKT1) in HUVECs was also quantified by Western Blot (WB). Results: Network pharmacological analysis revealed 283 target genes by ZYP in the treatment of RSA. Through protein-protein interaction (PPI) and topological analyses, a total of 57 key targets were identified as exerting central roles. Molecular docking proved that quercetin, β-carotene, resveratrol, and atractylenolide I had strong binding to AKT1 and VEGFA. Elevated levels of AKT1 and VEGFA protein expression were observed in WB analysis following intervention with the active ingredient. Discussion: The findings suggest that ZYP may ameliorate RSA by promoting angiogenesis at the maternal-fetal interface. Key active compounds were predicted to act via the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling pathway, as demonstrated by molecular docking and cell-based assays. Conclusion: This study illuminates the comprehensive therapeutic effects of ZYP in RSA treatment, providing a foundation for its further exploration.
zhou et al. (Mon,) studied this question.