Exosomal miR-199a-3p derived from follicular fluid modulates the immune microenvironment in polycystic ovary syndrome by regulating macrophage polarization
Abstract
BACKGROUND: Polycystic ovary syndrome (PCOS) is characterized by chronic low-grade inflammation of the ovaries. This disorder of the immune microenvironment is closely related to abnormal follicular development and ovulation disorders. This study aims to investigate whether the exosomes derived from the follicular fluid of PCOS patients regulate macrophage polarization by delivering miR-199a-3p, and further participate in the molecular mechanism of ovarian immune microenvironment imbalance. METHODS: Exosomes were extracted from the follicular fluid of both PCOS patients and control subjects. The expression levels of miR-199a-3p in exosomes were quantified through Quantitative reverse transcription PCR, while the concentrations of inflammatory cytokines were measured through ELISA. Bioinformatics analysis predicted mTOR as a potential target of miR-199a-3p. To investigate functional interactions, PCOS-derived exosomes were co-cultured with THP-1-derived macrophages, either in the presence or absence of miR-199a-3p mimics. After 48 h of incubation, the expression profiles of inflammatory factors(TNF-α, IL-6, IL-10 and TGF-β1), miR-199a-3p, M1/M2 macrophage polarization markers (CD86 and CD206), and mTOR were systematically analyzed. RESULTS: PCOS patients exhibited significantly upregulated miR-199a-3p in follicular fluid exosomes, along with elevated pro-inflammatory factors (TNF-α, IL-6) and decreased levels of anti-inflammatory cytokines (IL-10, TGF-β1). In an in vitro…
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