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.　©　The　Author(s)　2025.