Integrated multi-omics analysis and experimental verification reveal the involvement of the PI3K/Akt signaling pathway in myometrial fibrosis of adenomyosis

Adenomyosis (AM) is characterized as a chronic and progressive disorder with limited therapeutic strategies available. Myometrial fibrosis is a prominent pathological feature of AM, yet the underlying molecular mechanisms remain elusive. The present study conducted a comparative analysis using proteomics and metabolomics to investigate myometrial fibrosis and its underlying mechanisms. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was utilized to analyze adenomyotic and normal myometrial tissues from ten AM patients who underwent hysterectomy with myometrial fibrosis confirmed by Masson staining. This analysis established comprehensive proteomic and metabolomic profiles of AM patients and revealed widespread alterations in the proteome and metabolome within normal and fibrotic myometrium. Key proteins and signaling pathways linked to myometrial fibrogenesis were identified based on proteomic data. The integrated analysis showed significant associations between proteomic and metabolomic data and highlighted the critical role of the PI3K/AKT signaling pathway. Immunohistochemistry and Spearman's correlation analysis suggested a relationship between myometrial fibrosis and the metaplasia of myometrial stromal cells into myofibroblasts. Subsequent experiments identified crucial proteins and signaling pathways involved in myometrial fibrosis, indicating an association with the activation of the PI3K/AKT signaling pathway in myofibroblasts. Notably, PI3K/AKT inhibitors may contribute to the effective alleviation of myometrial fibrosis. This study is the first to demonstrate that myometrial fibrosis represents a critical pathological mechanism in AM through multi-omics methods and to elucidate the crucial role of the PI3K/AKT signaling pathway in this process. These findings provide valuable insights into the pathophysiology of AM and suggest antifibrotic treatment as a promising therapeutic strategy.