RESUMO
RESEARCH QUESTION: miRNA-339 participates in diseases with endothelial progenitor cell (EPC) dysfunction. What is the role of miRNA-339-5p in EPC of polycystic ovary syndrome (PCOS)? DESIGN: Clinical data were collected from 76 controls and 84 PCOS patients. Noradrenaline, asymmetric dimethylarginine (ADMA), advanced glycation end products (AGE) and silent information regulator 1 (SIRT1) in the serum were measured. The functions of EPC and the expressions of PI3K, AKT, SIRT1 and PGC-1α in EPC before and after transfection with miRNA-339-5p mimic or miRNA-339-5p inhibitor were compared. RESULTS: Serum concentrations of noradrenaline, ADMA and AGE were significantly higher (Pâ¯=â¯0.009, Pâ¯=â¯0.044, P < 0.001) and the SIRT1 concentration was significantly lower (P < 0.001) in PCOS patients, especially obese ones (Pâ¯=â¯0.034, Pâ¯=â¯0.032, P < 0.001, Pâ¯=â¯0.023) than in the control group. When compared with the controls, proliferation of the EPC was slightly lower (without a significant difference), the migration and tubular formation were significantly decreased (Pâ¯=â¯0.037, Pâ¯=â¯0.011), the expression of miRNA-339-5p in EPC was significantly higher (Pâ¯=â¯0.035) and the expressions of PI3K, AKT, SIRT1 and PGC-1α were significantly lower in the PCOS group (mRNA: Pâ¯=â¯0.033, Pâ¯=â¯0.027, Pâ¯=â¯0.027, Pâ¯=â¯0.032; protein: Pâ¯=â¯0.036, Pâ¯=â¯0.028, Pâ¯=â¯0.039, Pâ¯=â¯0.023). After transfection, the functions of EPC from PCOS patients were best in the miRNA-339-5p inhibitor group, and weakest in the miRNA-339-5p mimic group. The miRNA-339-5p inhibitor group had higher protein expressions of PI3K, AKT and SIRT1 but lower expression of PGC-1α in PCOS patients (P < 0.001, Pâ¯=â¯0.030, Pâ¯=â¯0.047, Pâ¯=â¯0.003). Similar results were obtained from the controls after transfection. CONCLUSION: Increased sympathetic excitation and damage to EPC were observed in PCOS patients, especially obese ones. Up-regulated miRNA-339-5p could inhibit the function of EPC by inhibiting the PI3K/AKT and SIRT1/PGC-1α signalling pathways.
Assuntos
Células Progenitoras Endoteliais , MicroRNAs , Síndrome do Ovário Policístico , Células Progenitoras Endoteliais/metabolismo , Feminino , Humanos , MicroRNAs/metabolismo , Norepinefrina , Obesidade , Fosfatidilinositol 3-Quinases/metabolismo , Síndrome do Ovário Policístico/genética , Síndrome do Ovário Policístico/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Sirtuína 1/genéticaRESUMO
The objective is to investigate the pathophysiological significance of Par3 and integrin ß1 with regard to the functionality of the endometrial luminal epithelium (LE). Design: laboratory study; setting: university research laboratory. Analysis involved endometrial aspirates and endometrial adenocarcinoma cells (HEC-1A) and endometrial carcinoma cells (RL95-2). We first examined the expression and localization of Par3 and integrin ß1 in HEC-1A cells and RL95-2 cells. Then we knocked down Par3 and integrin ß1 in HEC-1A cells and RL95-2 cells, respectively, and found that Par3/integrin ß1 affected embryo adhesion by regulating the intercellular tight junctions' (TJs') structure and thus the polarity of the endometrial LE. These findings were also confirmed in the endometrium specimens from human and mice. The main outcome measures were the expression and localization of Par3 and integrin ß1 in the endometrial epithelial cell lines and endometrium specimens and the regulations of Par3 and integrin ß1 on TJs, polarity, and embryo adhesion. Following the knockdown of Par3 in HEC-1A cells, there was a reduction in the complexity of the TJs and cell polarity, and the adhered blastocysts number was significantly increased. However, the reduction of integrin ß1 in RL95-2 cells resulted in effects that directly opposed those following the knockdown of Par3 in HEC-1A cells. Estrogen and progesterone reduced the expression of Par3 and promoted the expression of integrin ß1 in HEC-1A cells. Par3/integrin ß1 regulates embryo adhesion by regulating intercellular TJs' structure and polarity of endometrial LE under the action of ovarian hormones.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/genética , Proteínas de Ciclo Celular/genética , Polaridade Celular , Implantação do Embrião , Endométrio/fisiologia , Integrina beta1/genética , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Animais , Proteínas de Ciclo Celular/metabolismo , Linhagem Celular , Células Epiteliais/fisiologia , Feminino , Humanos , Integrina beta1/metabolismo , CamundongosRESUMO
Polycystic ovary syndrome (PCOS) is a systemic endocrine disease affecting women's reproductive health. Ovarian angiogenesis in PCOS patients is abnormal, manifested by increased ovarian stromal vascularization and upregulated proangiogenic factors such as vascular endothelial growth factor (VEGF). However, the specific mechanisms underlying these changes in PCOS remain unknown. In this study, we induced the adipogenic differentiation in preadipocyte 3T3-L1 cells and found that adipocyte-derived exosomes promoted proliferation, migration, tube formation, and VEGFA expression in human ovarian microvascular endothelial cells (HOMECs) by delivering miR-30c-5p. Mechanistically, dual luciferase reporter assay demonstrated that miR-30c-5p directly targeted the 3'- untranslated region (UTR) of suppressor of cytokine signaling 3 (SOCS3) mRNA. In addition, adipocyte-derived exosomal miR-30c-5p activated signal transducer and activator of transcription 3 (STAT3)/VEGFA pathway in HOMECs via targeting SOCS3. In vivo experiments indicated that tail vein injection of adipocyte-derived exosomes exacerbated endocrine and metabolic disorders and ovarian angiogenesis in mice with PCOS via miR-30c-5p. Taken together, the study revealed that adipocyte-derived exosomal miR-30c-5p promotes ovarian angiogenesis via the SOCS3/STAT3/VEGFA pathway, thereby participating in the development of PCOS.