From Constitution to Disease: MicroRNA Signatures for the Early Prediction and Targeted Prevention of Polycystic Ovary Syndrome

Na Shi; Shuhua Ji; Yan Zhang; Huiling Qu; Jinwei Hou; Longhuan Jiang; Hai-Ping Liu

doi:10.2147/IJWH.S565574

From Constitution to Disease: MicroRNA Signatures for the Early Prediction and Targeted Prevention of Polycystic Ovary Syndrome

Int J Womens Health. 2025 Dec 12:17:5389-5398. doi: 10.2147/IJWH.S565574. eCollection 2025.

Authors

Na Shi¹, Shuhua Ji¹, Yan Zhang¹, Huiling Qu¹, Jinwei Hou¹, Longhuan Jiang¹, Hai-Ping Liu¹

Affiliation

¹ Department of Reproductive Medicine, the 960th Hospital of the PLA Joint Logistics Support Force, Jinan, People's Republic of China.

Abstract

Background: The association between PCOS and the TCM concept of phlegm-damp constitution is well-documented, but their connection via miRNA remains unclear. This study sought to identify key miRNAs implicated in the transition from phlegm-damp constitution to PCOS and to elucidate their associated regulatory networks.

Methods: We conducted miRNA sequencing on granulosa cells from three groups: healthy controls with a neutral constitution (NP), healthy individuals with a phlegm-damp constitution (NB), and PCOS patients (PC). Principal component analysis (PCA) and differential expression analysis were utilized to pinpoint candidate miRNAs, which were subsequently validated using quantitative real-time PCR (qRT-PCR) in larger cohorts. Bioinformatics, including target prediction, protein-protein interaction (PPI) network construction, and pathway analysis, were employed to delineate the regulatory networks involved.

Results: PCA revealed distinct clustering patterns, with NB and PC groups exhibiting closer proximity compared to the NP group. Differential expression analysis identified 24 dysregulated miRNAs in the PC group (20 upregulated, 4 downregulated) and 14 in the NB group (11 upregulated, 3 downregulated), with both groups showing shared enrichment in metabolic, PI3K-Akt, and Ras pathways. Validation confirmed a progressive downregulation of hsa-miR-24-2-5p and hsa-miR-33a-5p across the NP, the NB, and PC groups (P<0.05), whereas hsa-miR-374b-5p exhibited differential expression exclusively between NB and PC groups. Network analysis identified five hub genes (EGFR, MYC, KRAS, CREB1, SMAD4) and nine risk pathways, including MAPK and Wnt signaling pathways. The constructed miRNA-hub gene-pathway network comprised five miRNA-hub gene pairs and thirteen functional modules.

Conclusion: These findings elucidate shared miRNA regulatory mechanisms between phlegm-damp constitution and PCOS, highlighting hsa-miR-24-2-5p and hsa-miR-33a-5p as pivotal mediators in this pathological transition. This provides molecular evidence for the TCM "constitution-disease correlation" theory and identifies potential targets for the prevention and treatment of PCOS.

Keywords: constitution-disease correlation; miRNA profiling; phlegm-damp constitution; polycystic ovary syndrome; traditional Chinese medicine.