Abstract: Objective  To investigate the molecular mechanism by which Yangxue Antai granules (YXATG) treat recurrent spontaneous abortion (RSA) through the regulation of ferroptosis. Methods  Network pharmacology was employed to screen the active ingredients and potential targets of YXATG by using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and other databases. The obtained targets were intersected with RSA-related and ferroptosis-related targets retrieved from databases such as GeneCards and FerrDb to identify core targets, followed by enrichment analysis. Molecular docking was performed to validate the binding affinity between core active ingredients and the targets. In vitro cell experiments were conducted for verification: drug-containing serum of YXATG was prepared from SD rats; the human choriocarcinoma cell line JEG-3 was used as a model, in which oxidative stress was induced by H²O² and intervened with the drug-containing serum; finally, Western blotting was used to detect the protein expression levels of the core targets TP53, PTGS2, and ICAM1.Results  A total of 88 active ingredients and 258 drug targets were screened. Twenty-two common targets among YXATG, RSA, and ferroptosis were identified, among which ICAM1, TP53, and PTGS2 emerged as the central targets. Enrichment analysis revealed that the targets were significantly enriched in pathways related to oxidative stress, inflammatory response, and lipid metabolism. Molecular docking confirmed that components such as diosgenin, stigmasterol, and quercetin exhibited good binding affinity to the core targets. In vitro experimental results showed that, compared with the control group, the protein expression levels of TP53, PTGS2, and ICAM1 were significantly increased in the H²O² model group (all P < 0.01); whereas, compared with the model group, intervention with YXATG-containing serum significantly inhibited the expression of all three proteins (all P < 0.01).Conclusion  YXATG may exert its therapeutic effect on RSA through the synergistic action of multiple components such as diosgenin and quercetin, targeting key molecules including TP53, PTGS2, and ICAM1, regulating their expression, thereby alleviating oxidative stress and inflammatory responses, inhibiting ferroptosis, and ultimately treating RSA.

Key words: Yangxue Antai granules, recurrent spontaneous abortion, ferroptosis, network pharmacology, molecular docking, Western blotting