lncRNA-miRNA-mRNA Network Analysis and Downstream ProteinsReveal the Mechanism of Bushen Huatan Prescription in the Treatment ofPolycystic Ovary Syndrome

Introduction: Polycystic Ovary Syndrome (PCOS) is a prevalent reproductive endocrine disorder. The underlying mechanisms, particularly the role of the long non-coding RNA (lncRNA)- miRNA-mRNA network, remain incompletely understood. The Bushen Huatan (BSHT) prescription shows clinical efficacy in treating PCOS, but its active components and their regulatory effects on this network are largely unknown. Method: Active components of the BSHT extract were analyzed using UHPLC-HR-MS. RNA and lncRNA sequencing were performed to identify differentially expressed lncRNAs, miRNAs, and mRNAs in PCOS rat models. A potential competing endogenous RNA (ceRNA) network was constructed and subsequently validated by Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, and qRT-PCR. Finally, Western blot analysis was used to evaluate the effect of BSHT on the expression of downstream insulin receptor proteins and signaling pathway proteins. Results: UHPLC-HR-MS identified 503 active ingredients in the BSHT extract. Compared to controls, PCOS rats exhibited 1181 differentially expressed lncRNAs and 2696 mRNAs. BSHT treatment significantly altered the expression of 336 lncRNAs and 678 mRNAs, and qRT-PCR showed that lncRNA XLOC-018280 expression was significantly downregulated by approximately 4-fold (P<0.01) in the MC group compared to the NC group, and this was significantly reversed by BSHT treatment. Western blot showed that INSR, IRS-1, IRS-2, PI3K, and p-AKT were differentially expressed. Conclusion: The Bushen Huatan prescription alleviates the endocrine and metabolic disorders of PCOS by regulating the lncRNA-miRNA-mRNA network and affecting the expression of downstream proteins INSR, IRS-1, IRS-2, PI3K, and p-AKT.