Journal of Shandong University (Health Sciences) ›› 2023, Vol. 61 ›› Issue (5): 51-58.doi: 10.6040/j.issn.1671-7554.0.2022.0891

• 临床医学 • Previous Articles    

Effects of human umbilical cord mesenchymal stem cell-derived exosomal miR-100 on inflammation of ovarian granulosa cells in polycystic ovary syndrome

ZHAO Yuanyuan1, LU Juntao2, WU Xiaohua1   

  1. 1. Center for Reproductive Medicine, The Fourth Hospital of Shijiazhuang(Gynecology and Obstetrics Hospital Affiliated to Hebei Medical University), Shijiazhuang 050011, Hebei, China;
    2. Laboratory of Pathology, Hebei Cancer Institute, The Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, Hebei, China
  • Published:2023-05-15

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Abstract: Objective To explore the effects of exosomal miR-100 derived from human umbilical cord mesenchymal stem cells(hUC-MSCs)on the inflammation of granulosa cells(GCs)in polycystic ovary syndrome(PCOS). Methods A total of 70 PCOS patients treated during Mar. 2021 and Dec. 2021 were enrolled as the PCOS group, and another 70 patients treated due to tubal factors were selected as the control group. The clinical characteristics and in vitro fertilization(IVF)outcomes were compared between the two groups. The expressions of pro-inflammatory cytokines TNF-α and IFN-γ in patients’ GCs were detected with real-time quantitative PCR(RT-qPCR). The risk factors influencing the occurrence of PCOS were analyzed with binary Logistic regression analysis. The expression of miR-100 in hUC-MSCs exosomes(exos)was verified with RT-qPCR. After the miR-100 mimics and miR-NC were synthesized and transfected into human ovarian granulosa cells(KGN), the expression of miR-100 was determined with RT-qPCR. After the miR-100 mimics and miR-NC were transfected into lipopolysaccharide(LPS)-induced KGN cells, the mRNA expressions of TNF-α and IFN-γ were detected with RT-qPCR. The target gene of miR-100 was verified with dual-luciferase reporter gene assay. After miR-100 was overexpressed in LPS-induced KGN cells, the effector molecular of Toll-like receptor-4/nuclear factor kappa B(TLR4/NF-κB)signaling pathways, including TLR4, p-IκBa, IκBa, p-p65 and p65, were detected with Western blotting. Results PCOS group had higher body mass index(BMI), luteinizing hormone(LH), estradiol(E2), testosterone(T), homeostasis model assessment for insulin resistance index(HOMA-IR), antral follicle count(AFC), and number of retrieved oocytes than the control group, whereas lower MII oocyte rate and 2PN fertilization rate(P<0.001, P<0.001, P=0.020, P<0.001, P<0.001, P<0.001, P<0.001, P<0.001, P<0.001). The PCOS group had significantly higher expressions of TNF-α and IFN-γ than the control group(P<0.001). Binary Logistic regression analysis revealed that LH, HOMA-IR, TNF-α and IFN-γ were risk factors influencing the occurrence of PCOS(P=0.031, 0.001, 0.030, 0.005). RT-qPCR showed that miR-100 was enriched in hUC-MSCs-exos. Overexpression of miR-100 significantly inhibited LPS-induced increase of the mRNA expressions of TNF-α and IFN-γ(P=0.041, P<0.001). Dual-luciferase reporter gene assay showed that miR-100 directly targeted TLR4 mRNA 3’UTR and inhibited its expression(P<0.001). Western blotting showed that miR-100 efficiently inhibited LPS-induced elevation of the protein expressions of TLR4, p-IκB and p-p65(P<0.001, P<0.001, P=0.049), but the protein expression of IκB significantly increased(P<0.001). Conclusion The increased expressions of TNF-α and IFN-γ are risk factors influencing the occurrence of PCOS. Exosomal miR-100 derived from hUC-MSCs inhibits the inflammation of granulosa cells via TLR4/NF-κB signaling pathway in PCOS.

Key words: Polycystic ovary syndrome, Human umbilical cord mesenchymal stem cell, Exosomes, MicroRNAs, Granulosa cells, Inflammation

CLC Number: 

  • R711.75
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