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.

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.

Objective To investigate the epidemiologic factors associated with the age of natural menopause and menopausal symptoms in a large population at age of 40 to 65 years in Jiangsu Province.Methods From May 2010 to Oct.2011,a total of 20 275 women (40 to 65 years) attending health examination in Jiangsu Province were enrolled in this cross-sectional study.A structured questionnaire was used to collect data of demographics,menopausal status,chronic diseases,reproductive history.Also the menopausal symptoms were evaluated by Kupperman menopause index (KMI).Cox proportional hazards regression model and Logistic regression were performed to identify risk factors for earlier age of natural menopause and menopausal symptoms,respectively.Results The overall median age at natural menopause in Jiangsu women was 50 years.Lower educational level,poor economic status,lower body mass index (BMI),age at menarche less than 14 years,nulliparity and smoking were associated with earlier onset of natural menopause (P ＜ 0.05).The most frequently symptoms in perimenopausal women were fatigue (46.84％,1880/4014),insomnia (44.67％,1793/4014) and muscle/joint pain (43.80％,1758/ 4014),while sexual problems (57.06％,3463/6070),muscle/joint pain (53.30％,3235/6070) and insomnia (51.03％,3097/6070) were predominant symptoms in postmenopausal women.After adjusting for confounding factors,it was revealed that women with poor educational background,low income,divorce,higher BMI,higher parity,and smoking presented positive correlation with menopausal symptoms (P ＜ 0.05).Conclusions The study suggested that an estimate of median age at natural menopause were 50 years in Jiangsu women.The main factors contributing to earlier onset of menopause and menopausal symptoms were lower educational level,poor economic status,and smoking.Moreover,there were different menopausal symptoms between perimenopausal and postmenopausal women,which provided the important insights for physicians to prevent and treat menopause symptoms in their clinical practice.