Background The aetiology and pathogenesis of adenomyosis remain unclear. This study utilised untargeted metabolomics to explore the aberrant
Background The aetiology and pathogenesis of adenomyosis remain unclear. This study utilised untargeted metabolomics to explore the aberrant amino acid metabolism in adenomyosis. Among the studied metabolites, hydroxyproline is known to promote the proliferation and invasion of tumour cells. Due to detection limitations, hydroxyproline levels were inferred via the expression of prolyl 4-hydroxylase subunit alpha 1 (P4HA1), a crucial enzyme that catalyses the conversion of proline to hydroxyproline.Methods Endometrial metabolomic analysis was performed using gas chromatography-mass spectrometry (GC-MS) in 15 and 20 patients with and without adenomyosis. The expression levels of P4HA1 mRNA and protein were detected using quantitative PCR (qPCR), western blotting, and immunohistochemistry. In vitro, Cell Counting Kit-8, and 5-Ethynyl-2′-deoxyuridine (EdU) assays were used to investigate the effect of hydroxyproline on the proliferation of human endometrial stromal cells (hESCs). Additionally, a wound-healing assay was conducted to examine the effect of hydroxyproline on hESCs migration.Results In total, 22 differentially expressed metabolites were identified in the adenomyosis group compared to the control group. qPCR results demonstrated that P4HA1 mRNA expression levels were significantly higher in the adenomyosis group than in the control group (p
