Paeonol alleviates intrauterine adhesion by suppressing endometrial stromal cell activation via AKT1, based on network pharmacology

CONTEXT: Intrauterine adhesion (IUA) causes secondary infertility and recurrent miscarriage, yet therapeutic options remain limited. Paeonol is widely used for gynecological diseases, but its precise role in IUA therapy is unclear. AIMS: This study aims to explore the molecular mechanism of paeonol in IUA. METHODS: Network pharmacology identified paeonol's potential targets and core proteins via a protein-protein interaction network. Bioinformatic pathway enrichment analyses and molecular docking elucidated mechanisms and validated binding. The optimal concentration and treatment time of paeonol were screened using Cell Counting Kit-8 (CCK-8) assays. In a transforming growth factor (TGF)-β-induced activated primary endometrial stromal cell (ESC) model, paeonol's effects on proliferation and migration were evaluated using CCK-8, wound healing, and Transwell assays. Expression levels of AKT1 and collagen synthesis-related markers were assessed by Western blotting. Protein levels of type I collagen and vimentin were assessed by immunofluorescence staining. In a IUA rat model, the in vivo efficacy of paeonol in alleviating IUA via AKT1 regulation was validated based on pathological, inflammatory, and reproductive parameters. KEY RESULTS: Paeonol targets were significantly enriched in the phosphoinositide 3-kinaseK-AKT pathway. In vitro, paeonol blocked TGF-β-induced activation of primary ESC, reducing cell viability and migration. Mechanistically, paeonol antagonised TGF-β-induced ESC activation by suppressing AKT1, thereby attenuating IUA. In vivo, paeonol alleviated pathological changes of endometrial tissues, reduced fibrosis, and suppressed inflammation in IUA rats. CONCLUSION: Paeonol blocks AKT1 activation to suppress inflammation and fibrosis, ultimately slowing IUA progression. IMPLICATIONS: This study provides a new theoretical basis for paeonol use in IUA therapy.