Reproductive biomedicine online · 2025
Differential expression of extracellular vesicle microRNA and granulosa cell mRNA in polycystic ovary syndrome with insulin resistance.
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Ju Wenhan, Pan Keying, Zhang Qianwen, Wang Yue, Zhao Shuai, Zhang Jinfu, Lian Fang
Аннотация
RESEARCH QUESTION: How does the microRNA (miRNA)-mRNA regulatory network in polycystic ovary syndrome (PCOS) with insulin resistance (PCOS-IR) manifest, based on miRNA sequencing of follicular fluid-derived extracellular vesicles and mRNA sequencing of granulosa cells from clinical patients?
DESIGN: Follicular fluid and granulosa cells were collected from patients with PCOS-IR and PCOS and healthy controls. Differentially expressed miRNA (DEmiR) and differentially expressed mRNA (DEmRNA) among the three groups were identified and analysed using high-throughput sequencing. DEmiR expression was validated by quantitative real-time (qRT)-PCR. Finally, miRNA-mRNA interaction networks were constructed to identify follicular fluid-derived extracellular miRNA and target mRNA in granulosa cells potentially associated with PCOS-IR pathogenesis.
RESULTS: Comparison between the three groups identified 43 DEmiR in follicular fluid-derived extracellular vesicles. DEmiR were primarily involved in the regulation of metabolic pathways and signalling pathways. qRT-PCR confirmed the differential expression of four DEmiR (miR-324-5p, miR-345-5p, miR-500a-3p and let-7i-3p) in follicular fluid-derived extracellular vesicles, consistent with the sequencing results. A total of 797 DEmRNA were identified in granulosa cells, primarily involved in biological processes such as transcriptional regulation, lipid metabolism, apoptosis and immunity. Construction of the follicular fluid-derived miRNA-granulosa cell mRNA network identified 174 potential target genes mainly involved in the regulation of the PI3K-Akt signalling pathway, endocytosis, calcium signalling pathway, focal adhesion and autophagy.
CONCLUSIONS: This study presents the first construction of a follicular fluid-derived extracellular vesicle miRNA and granulosa cell mRNA dysregulation network in women with PCOS-IR, highlighting its potential role in regulating the pathogenesis of PCOS-IR.
DESIGN: Follicular fluid and granulosa cells were collected from patients with PCOS-IR and PCOS and healthy controls. Differentially expressed miRNA (DEmiR) and differentially expressed mRNA (DEmRNA) among the three groups were identified and analysed using high-throughput sequencing. DEmiR expression was validated by quantitative real-time (qRT)-PCR. Finally, miRNA-mRNA interaction networks were constructed to identify follicular fluid-derived extracellular miRNA and target mRNA in granulosa cells potentially associated with PCOS-IR pathogenesis.
RESULTS: Comparison between the three groups identified 43 DEmiR in follicular fluid-derived extracellular vesicles. DEmiR were primarily involved in the regulation of metabolic pathways and signalling pathways. qRT-PCR confirmed the differential expression of four DEmiR (miR-324-5p, miR-345-5p, miR-500a-3p and let-7i-3p) in follicular fluid-derived extracellular vesicles, consistent with the sequencing results. A total of 797 DEmRNA were identified in granulosa cells, primarily involved in biological processes such as transcriptional regulation, lipid metabolism, apoptosis and immunity. Construction of the follicular fluid-derived miRNA-granulosa cell mRNA network identified 174 potential target genes mainly involved in the regulation of the PI3K-Akt signalling pathway, endocytosis, calcium signalling pathway, focal adhesion and autophagy.
CONCLUSIONS: This study presents the first construction of a follicular fluid-derived extracellular vesicle miRNA and granulosa cell mRNA dysregulation network in women with PCOS-IR, highlighting its potential role in regulating the pathogenesis of PCOS-IR.