Exploring the Therapeutic Potential of 3‐Epioleanolic Acid in Steroid‐Induced Osteonecrosis of the Femoral Head Through Network Pharmacology

ABSTRACT This study explores the potential mechanisms by which 3‐epioleanolic acid, a bioactive compound from Hedyotis diffusa , facilitates the repair of steroid‐induced osteonecrosis of the femoral head (SONFH), through the integration of network pharmacology and bioinformatics approaches. Using SwissTargetPrediction and GeneCards databases, key overlapping targets, such as CYP19A1, TERT, and PTPN6, were identified, suggesting their involvement as potential therapeutic mediators. Molecular docking simulations confirmed strong binding affinities between 3‐epioleanolic acid and these targets. Functional enrichment analysis indicated that the compound may exert therapeutic effects by modulating pathways associated with steroid binding and glucocorticoid metabolism. Transcriptomic data analysis further validated the differential expression of CYP19A1, TERT, and PTPN6 during SONFH progression, underscoring their critical roles in disease development. The CCK‐8 assay results revealed that 80 μM 3‐epiursolic acid alleviated the proliferation inhibition induced by SONFH. Western blotting and RT‐qPCR analyses demonstrated that treatment with 3‐epiursolic acid significantly upregulated the expression levels of CYP19A1 and TERT, while markedly downregulating PTPN6 expression. These findings were consistent with the results of bioinformatics analysis, further confirming the regulatory effects of 3‐epiursolic acid on these potential targets. Collectively, these findings provide mechanistic insights into the therapeutic potential of 3‐epioleanolic acid for SONFH and support its application as a promising candidate for disease intervention and progression monitoring.