RuAn Tablets Relieve Mammary Gland Hyperplasia by Regulating PGE2 Synthesis

Introduction: RuAn Tablets (RATs) are a traditional Chinese formulation used for the treatment of mammary gland hyperplasia (MGH), one of the most prevalent gynecological diseases. However, the complexity of RATs composition has hindered the elucidation of the mechanism. Methods: In this study, we employed an integrated approach combining ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS), network pharmacology, and experimental validation to identify the active components of RATs and explore their potential mechanisms in alleviating MGH. Results: Through UHPLC-MS analysis and database screening (SymMap and TCMSP), four active compounds were screened, including Betulinic acid (BA), Dioscin, Icariside I, and Neohesperidin. These compounds were predicted to interact with 321 potential targets, of which 22 were identified as key targets for RATs in MGH intervention. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses revealed that these targets were primarily associated with prostaglandin synthesis and metabolism, particularly the synthesis of prostaglandin E2 (PGE2). Molecular docking studies demonstrated strong binding affinities between the active compounds and core target proteins, including PTGS2, EGFR, and TP53. Experimental validation further confirmed that RATs significantly reduced PGE2 levels in mammary proliferating cells by over 60%. Discussion: Integrating findings from network pharmacology and experimental validation suggests that RATs exert their therapeutic effects on MGH primarily by regulating PGE2 synthesis. Conclusion: In summary, this study provides a comprehensive understanding of the molecular mechanisms underlying RATs' efficacy and highlights their potential as a therapeutic agent for MGH.