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.