Mechanism of serum exosomal miR-17-5p on regulation of Treg cells in premature ovarian insufficiency
Objective To explore the involvement of serum exosomal miR-17-5p in the pathogenesis of regulatory T(Treg)cells deficiency in premature ovarian insufficiency(POI)patients.Methods In a case-control study,32 Chinese Han women with POI and 32 Chinese Han women with regular menstrual cycles(healthy control,HC)attending the Department of Gynecological Endocrinology,the First Affiliated Hospital of Nanjing Medical University from January 2019 to December 2020 were recruited.The relative expressional level of serum exosomal miR-17-5p was confirmed by reverse transcription quantitative polymerase chain reaction(RT-qPCR).To understand how POI-exosome affects Treg cells,peripheral blood mononuclear cells(PBMC)of healthy volunteer were transfected with miR-17-5p,and cultured with PBS as the control or with exosomes derived from POI women or HC for 72 h.Subsequently,to evaluate the functionality of the miR-17-5p upregulated in POI-exosome,native CD4+ T cells from healthy donors were sorted by magnetic beads and induced into induced Treg(iTreg)cells in vitro,and the induction efficiency,the proliferation and apoptosis rates of iTreg cells were detected after transfection with miR-17-5p mimic/inhibitor.TargetScan,miRDB,miRTarBase and miRWalk were used to predict targets gene of miR-17-5p.Results The expression level of serum exosomal miR-17-5p was markedly up-regulated in POI compared with HC(P<0.001).Furthermore,after culture with POI-exosome[(0.93±0.40)%],the frequency of Treg cells was significantly reduced as compared with that culture with PBS[(3.77±0.89)%,P=0.005].Subsequently,the frequency of Treg cells in miR-17-5p mimic group[(4.30±1.91)%]was significantly lower than that in negative control group[(11.97±1.82)%,P=0.007].In contrast,both Th1 and Th17 did not be affected(both P>0.05).The induction efficiency of iTreg cells did not be influenced by transfection with miR-17-5p(both P>0.05),however,the iTreg cells apoptosis rate in miR-17-5p mimic group[(16.31±1.27)%]was significantly higher than that in negative control group[(13.01±1.80)%,P=0.035],while apoptosis rate was significantly lower[(7.97±0.71)%]and proliferation rate was higher[(88.83±0.25)%]in miR-17-5p inhibitor group than those in negative control group[(11.42±0.23)%,P=0.001;(87.60±0.70)%,P=0.045].Transforming growth factor beta receptor 2 was most likely targets gene of miR-17-5p.Conclusion The exosomal miRNA profile of patients with POI differed from that of HC,and miR-17-5p,which is markedly increased in POI patients,decreased the frequency of Treg cells by inhibiting iTreg cell proliferation and promoting iTreg cells apoptosis.Thus,our study suggested that exosomal miR-17-5p promoted iTreg cells apoptosis and resulted in Treg cells deficiency in POI pathogenesis.
ExosomesPremature ovarian insufficencyRegulatory T cellsmiR-17-5p
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