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@#Objective To evaluate the effect of follicular fluid(FF)exosomal miRNAs on follicular dysplasia in patients with polycystic ovary syndrome(PCOS)mediated by glycolysis pathway of granulosa cells(GCs),and to explore the mechanism. Methods Three PCOS infertile patients and three non-PCOS infertile patients were recruited. The baseline hormone levels of the two groups were measured before ovulation induction. The bilateral FF was obtained by puncture after short-acting and long-term ovulation induction,and the exosomes were collected by ultracentrifugation and identified by transmission electron microscopy. The total exosomal RNA was extracted by Trizol method to construct the library,which was compared to the reference genome GRCh38 for statistical analysis after miRNA sequencing and quality control processing. Clustering Profiler R package was used to implement GO annotation analysis and KEGG pathway analysis of the differentially expressed genes(DEGs),and Omnipath software for miRNAs interaction analysis. A total of 16 miRNA were randomly selected and detected by qPCR to verify the accuracy of the miRNA sequencing results. Results Compared with the non-PCOS group,luteinizing hormone(LH),anti-Muerian hormone(AMH),testosterone and antral follicle counts in PCOS group increased significantly(t = 2. 479 ～ 9. 163,each P < 0. 05). The exosomes of FF in both groups showed the cup-shaped vesicles with clear edge and light staining in the center,with the diameters of 100 — 150 nm and intact structure,and the concentration was about 8 × 1010particles/mL. A total of 928 miRNAs were detected by miRNA sequencing. Compared with the non-PCOS group,59 differentially expressed miRNA(DEmiRNA)were screened out in exosomes of POCS group,of which 31 were up-regulated and 28 were down-regulated. The differential trend of gene expression detected by qPCR was highly similar to that of miRNA sequencing. In FF exosomes of PCOS patients,the glycolysis efficiency and apoptosis of GCs were significantly changed by miRNA regulating mRNA. PKM,PFKL and HK2 were the key target genes for miRNA to regulate GCs glycolysis,and SLC2A1 was the key target gene for miRNA to regulate GCs apoptosis. Conclusion The miRNAs in FF exosomes of PCOS patients can weaken the glycolysis of GCs while accelerate the apoptosis,thus reducing the production of ATP and lactic acid,resulting in follicular dysplasia.
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ObjectiveTo investigate the mRNA expression levels of various aquaporins （AQPs） in luteinized granulosa cells from follicles of different diameters. MethodsFrom March 25， 2022 to September 23， 2022 in our reproductive medicine center， 48 women undergoing in-vitro fertilization （IVF） were enrolled and divided into the antagonist group and the agonist group according to the ovarian stimulation protocol. Follicular fluid samples were collected on the day of oocyte pick-up and granulosa cells were extracted from follicles of different diameters： small （<13 mm）， medium （13~18 mm） and large （≥18 mm）. After RNA quantification， 22 cases （66 samples） were included for analysis and mRNA expression levels of AQPs were compared among the three follicle groups. ResultsThe mRNA expression of aquaporin 2 （AQP2） in luteinized granulosa cells increased with the increase of follicle diameter （linear trend P = 0.004） and the difference was statistically significant between two groups of large and small follicles （P = 0.017）. Statistical difference was found in the antagonist group （P = 0.049 6）， but not in the agonist group （P = 0.108）. ConclusionThe mRNA level of AQP2 in luteinized granulosa cells increases with the increase of follicle diameter and its expression is related to the ovarian stimulation protocol， suggesting that AQP2 may play a role in follicle growth and follicular fluid formation， and its mRNA expression level may be regulated by follicle stimulating hormone （FSH） and luteinizing hormone （LH）.
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Objective To investigate whether the metabolites in follicular fluid can be used as indicators for predicting oocyte quality and embryos early development in patients with polycystic ovary syndrome (PCOS). Methods The study subjects were divided into four groups: lean control (LC) 30 cases, overweight (OW) 13 cases, lean PCOS (LP) 26 cases, and overweight PCOS (OP) 26 cases based on the Chinese criteria for body mass index (BMI) categories. The metabolic variance of follicular fluid from PCOS and controls was performed by liquid chromatography-mass spectrometry (LC-MS), and clinical characteristics, oocyte outcomes and differences of metabolites in follicular fluid of those patients have been compared by ANOVA. Furthermore, Pearson's correlation analysis was used to explore the correlation between oocyte fertilization rate, top-quality embryos rate and follicular metabolites. Results Compared to the LC group, oocytes retrieved and top-quality embryos rate were significantly increased, while the mature oocytes rate and fertilization rate decreased significantly in the OP group. A total of 236 metabolites were identified and quantified by metabolomics in follicular fluid. Compared with LC and OP groups, 19 metabolites showed statistically significant differences in PCOS group. Additionally, 7 metabolites were significant correlated with the fertilization rate or top-quality embryo rate respectively, most of which were lysophospholipids. Conclusion Several metabolites in the follicular fluid are significantly different between PCOS and healthy women. Among these, lysophospholipids are crucial for oocyte quality and early embryonic development, may serve as potential markers to evaluate the oocyte quality in PCOS patients.
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Objective: To identify the differential proteins from follicular fluid of patients with polycystic ovarian syndrome (PCOS) employing protein chip array, and provide a theoretical basis for further study of PCOS. Methods: AAH-BLG-507 array was used to test the follicular fluid of 6 PCOS patients with normal body mass index (BMI) and 6 non-PCOS controls with age and BMI-matched. The differential proteins were identified and bioinformatics analysis was carried out by functional annotation analysis, KEGG pathway analysis and protein-protein interaction network analysis. Results: Compared with non-PCOS controls, 25 up-regulated proteins and 49 down-regulated proteins were identified from follicular fluid of patients with PCOS. The classification of differential proteins based on molecular function revealed that they were mainly involved in cytokine activity and chemokine activity. KEGG pathway analysis was performed and pathways associated with cytokine-cytokine receptor interaction and chemokine signaling pathway were significantly enriched. The core proteins filtered by protein-protein interaction network analysis were mainly chemokines and their receptors. Conclusion: Protein chip array can be used to establish the differential expressed protein profile from follicular fluid of patients with PCOS. The cytokinecytokine receptor interaction and chemokine signaling pathway may be involved in the pathogenesis of PCOS.
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Extracellular vesicles (EVs) refer to bilayer membrane transport vesicles secreted by cells. EVs can take macromolecules from cells and transfer them to receptor cells. Among these macromolecular substances, the most studied are microRNAs (miRNAs). miRNA is non-coding RNA involved in the regulation of gene expression. It has been confirmed that there are different non-coding RNAs in mammalian follicular fluid EVs. EVs carrying miRNA can act as an alternative mechanism for autocrine and paracrine, affecting follicular development. This paper systematically introduced the kinds, characteristics and methods of isolation and identification of EVs, focusing on the effects of EVs and miRNAs on follicular development, including early follicular development, oocyte maturation, follicular dominance and effects on granulosa cell function. At the same time, the authors prospected the future research of EVs and microRNAs in follicular fluid, and provided ideas and directions for the research and application of EVs and miRNA functions in follicular fluid.
Subject(s)
Animals , Female , Extracellular Vesicles , Metabolism , Follicular Fluid , Chemistry , Granulosa Cells , MicroRNAs , Pharmacology , OogenesisABSTRACT
OBJECTIVE@#To investigate the correlation between soluble receptor for advanced glycation end products (sRAGE) level in the follicular fluid and ovarian responsiveness in non-PCOS patients undergoing controlled ovarian hyperstimulation.@*METHODS@#Ninety non-PCOS patients underwent IVF/ICSI using a short-acting long protocol for ovarian stimulation with a GnRH agonist. For each patient, the level of sRAGE in the follicular fluid was measured by enzyme linked immunosorbent assay (ELISA), and the data including the clinical baseline state, hormone level, number of oocytes obtained and the fertilization rate were collected.@*RESULTS@#Follicular fluid sRAGE level showed significant negative correlations with basal FSH level (=0.0036) and Gn dose ( 15) than in cases with oocytes obtained within the range of the target numbers (7-15) and below the target number (< 7) ( < 0.0001 and =0.0012, respectively).@*CONCLUSIONS@#Follicular fluid sRAGE level can reflect ovarian reserve function in non-PCOS patients, the number of oocytes obtained and the fertilization rate, and can thus predict ovarian responsiveness during controlled hyperstimulation in nonPCOS patients.
Subject(s)
Female , Humans , Fertilization in Vitro , Follicular Fluid , Ovarian Hyperstimulation Syndrome , Receptor for Advanced Glycation End ProductsABSTRACT
Objective: To explore the relationship of 11β-hydroxysteroid dehydrogenases (11β-HSDs) expression in the ovarian granulosa cells and cortisol regeneration in the ovary of polycystic ovary syndrome (PCOS). Methods: In accordance with the revised Rotterdam inclusion/exclusion criteria (2003), 24 patients in the PCOS group were included, meanwhile 21 patients in the control group were included. Follicular fluid and luteinized granulosa cells were collected from patients underwent in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) in the Center for Reproductive Medicine of Renji Hospital. Enzyme linked immunosorbent assay (ELISA) was used to detect the cortisol concentration and the cortisone levels of follicular fluid. Real-time quantitative PCR (RT-qPCR) was used to measure 11β-HSDs abundance in the luteinized granulosa cells. The expression of 11β-HSD1 and 11β-HSD2 in the luteinized granulosa cells were verified by immunofluorescence. The correlation of local cortisol level in the ovary with 11β-HSDs expression in the luteinized granulosa cells was further analyzed. Results: Compared with the control group, the cortisol level in the follicular fluid was elevated in the PCOS group [(477.3±35.3) nmol/L vs (292.0±36.4) nmol/L, P=0.014], while no change was observed in the cortisone levels in two groups. Both 11β-HSD1 and 11β-HSD2 were detected in the luteinized granulosa cells. Compared with the control group, the 11β-HSD1 mRNA abundance (P=0.033) and reductase activity (P=0.006) were increased significantly in the PCOS group, and there was no significant difference in the 11β-HSD2 expression and oxidase activity between two groups. Consistently, Pearson correlation analysis showed that the 11β-HSD1 but not 11β-HSD2 mRNA levels in the luteinized granulosa cells was correlated with the cortisol level in the follicular fluid (r2=0.347 6, P=0.001). Conclusion: Elevation of local cortisol in the ovary of PCOS patients may be related to the increased expression of 11β-HSD1 and reductase activity in the ovarian granulosa cells.
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Objective · To extract and identify exosomes in follicular fluid from patients with polycystic ovary syndrome in order to determine the existence of exosomes in follicular fluid, to isolate and extract miRNAs in exosomes, and to detect relative expression of miRNAs. Methods · Exosomes in follicular fluid were collected with membrane affinity chromatography and their size and morphology were observed with the transmission electron microscope. Exosome protein markers CD63 and CD81 were detected with flow cytometry. miRNAs in purified exosomes were extracted and expressions of miR-125b, miR-19b, and miR-222 were measured with TaqMan real-time PCR. Results · Exosomes in follicular fluid were circular or elliptic under the transmission electron microscope with diameters of around 30-100 nm. They had complete membrane structure and contained low density matter. Flow cytometry showed that CD63 and CD81 were positively expressed in exosomes. Real-time PCR detected expressions of miR-125b, miR-19b, and miR-222. Conclusion · Exosomes can be collected in follicular fluid from patients with polycystic ovary syndrome. Transmission electron microscopy and flow cytometry can be used to identify exosomes in follicular fluid. miR-125b, miR-19b, and miR-222 can be detected in exosomes.
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OBJECTIVE: The aim of this study was to assess the changes of follicular fluid (FF) and serum levels of cerebellin precursor protein 1 (cbln1) and betatrophin in patients with polycystic ovary syndrome (PCOS) undergoing in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) with a gonadotropin-releasing hormone (GnRH) antagonist protocol. METHODS: Twenty infertile women with PCOS and 20 control women diagnosed as poor responders undergoing ovarian stimulation with a GnRH antagonist were included. Blood samples were obtained during ovum pick-up. Follicular fluid from a dominant follicle was collected from the subjects. Using enzyme-linked immunosorbent assays, FF and serum levels of cbln1 and betatrophin were measured in both groups of participants. Metabolic and hormonal parameters were also determined and correlated with each other. RESULTS: Both groups of women had similar serum and FF betatrophin levels (55.0±8.9 ng/mL vs. 53.1±10.3 ng/mL, p=0.11). The serum and FF betatrophin levels of poor responders were found to be similar (49.9±5.9 ng/mL vs. 48.9±10.7 ng/mL, p=0.22). Conversely, the FF cbln1 levels of PCOS women were found to be significantly higher than the serum cbln1 levels (589.1±147.6 ng/L vs. 531.7±74.3 ng/L, p<0.02). The FF cbln1 levels of control participants without PCOS were significantly higher than their serum cbln1 levels (599.3±211.5 ng/L vs. 525.3±87.0 ng/L, p=0.01). Positive correlations were detected among body mass index, insulin resistance, serum insulin, total testosterone, and betatrophin levels in the PCOS group. CONCLUSION: Follicular fluid betatrophin and cbln1 concentrations may play a pivotal role on follicular growth in PCOS subjects undergoing IVF/ICSI with an antagonist protocol.
Subject(s)
Female , Humans , Body Mass Index , Enzyme-Linked Immunosorbent Assay , Fertilization in Vitro , Follicular Fluid , Gonadotropin-Releasing Hormone , In Vitro Techniques , Insulin , Insulin Resistance , Ovulation Induction , Ovum , Polycystic Ovary Syndrome , Sperm Injections, Intracytoplasmic , Spermatozoa , TestosteroneABSTRACT
OBJECTIVE: Hyperstimulation methods are broadly used for in vitro fertilization (IVF) in patients with infertility; however, the side effects associated with these therapies, such as ovarian hyperstimulation syndrome (OHSS), have not been well studied. N-glycoproteomes are subproteomes used for the remote sensing of ovarian stimulation in follicular growth. Glycoproteomic variation in human follicular fluid (hFF) has not been evaluated. In this study, we aimed to identify and quantify the glycoproteomes and N-glycoproteins (N-GPs) in natural and stimulated hFF using label-free nano-liquid chromatography/electrospray ionization-quad time-of-flight mass spectrometry. METHODS: For profiling of the total proteome and glycoproteome, pooled protein samples from natural and stimulated hFF samples were selectively isolated using hydrazide chemistry to obtain the total proteomes and glycoproteomes. N-GPs were validated by the consensus sequence N-X-S/T (92.2% specificity for the N-glycomotif at p<0.05). All data were compared between natural versus hyperstimulated hFF samples. RESULTS: We detected 41 and 44 N-GPs in the natural and stimulated hFF samples, respectively. Importantly, we identified 11 N-GPs with greater than two-fold upregulation in stimulated hFF samples compared to natural hFF samples. We also validated the novel N-GPs thyroxine-binding globulin, vitamin D-binding protein, and complement proteins C3 and C9. CONCLUSION: We identified and classified N-GPs in hFF to improve our understanding of follicular physiology in patients requiring assisted reproduction. Our results provided important insights into the prevention of hyperstimulation side effects, such as OHSS.
Subject(s)
Female , Humans , Chemistry , Complement System Proteins , Consensus Sequence , Fertilization in Vitro , Follicular Fluid , In Vitro Techniques , Infertility , Mass Spectrometry , Ovarian Hyperstimulation Syndrome , Ovulation Induction , Physiology , Proteome , Proteomics , Reproduction , Sensitivity and Specificity , Thyroxine-Binding Globulin , Up-Regulation , Vitamin D-Binding ProteinABSTRACT
Objective To investigate the mechanism by which follicular fluid from endometriosis (EM) patients blocks mouse oocyte maturation and the effect of Neiyi Recipe-medicated serum on the effect of the follicular fluid. Methods A total of 20 follicular fluid samples from infertility patients due to EM (n=10) or fallopian tube factors (control group, n=10) were clinically obtained. Neiyi Recipe-medicated serum was obtained by treating mice with intragastric administration, and were divided into 4 medium groups: Group A (control), group B (control follicular fluid), group C (EM-follicular fluid) and group D (EM-follicular fluid + Neiyi Recipe-medicated serum). The mouse germinal vesicle (GV) oocytes were cultured in the above 4 types of media for in vitro maturation. And the maturity rates of oocytes in different groups were calculated and the concentrations of reactive oxygen species in the cells were analyzed by fluorescence staining and laser scanning confocal microscope. Results The maturity rate of oocytes in group C (42.5%) was significantly lower than those in group A (81.7%), group B (56.3%) and group D (51.0%) (P<0.05). The average fluorescence intensity of group C (0.056 8±0.025 1) was significantly higher than those of group A (0.005 1±0.014 8, P<0.01) and group B (0.037 1±0.010 2, P<0.05), but with no significant difference found when compared with that of group D (0.050 4±0.007 0). Conclusion EM-follicular fluid can block the maturation of mouse oocytes, which may be due to the enhanced oxidative stress in oocytes caused by EM-follicular fluid. Neiyi Recipe-medicated serum can improve the blocking effect of EM-follicular fluid on mouse oocyte development.
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Objective To investigate the relationship of nerve growth factor (NGF) between serum and follicular fluid in patients with polycystic ovary syndrome (PCOS), and its effect on pregnancy outcomes. Methods From December 2011 to November 2012, 65 PCOS patients suffered with in vitro fertilization-embryo transfer (IVF-ET) were included in PCOS group and 31 non-PCOS patients with IVF-ET were used as control group. The expressions of NGF in serum and follicular fluid were detected by ELISA on the injection day of chorionic gonadotropin (HCG). The expression levels of NGF in serum and follicular fluid were compared between PCOS group and control group. Results The NGF levels in serum[(14.38±0.42) ng/L] and follicular fluid[(9.61±0.49)ng/L] were significantly higher in PCOS group than those of control group[(11.39±0.38) ng/L and (7.55 ± 0.40)ng/L]. In PCOS group, NGF levels in serum [(14.22 ± 0.35)ng/L] and follicular fluid [(9.30 ± 0.31)ng/L] were significantly lower in pregnant group than those of non-pregnant group[(14.51±0.43)ng/L, (9.86±0.46)ng/L, P<0.01]. Conclusion The increased levels of NGF in serum and follicular fluid of PCOS patients contribute to the pathogenesis of PCOS. The high expression of NGF in serum and follicular fluid may be harmful to pregnancy in PCOS patients.
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AIM:To retrospectively analyze the lipid metabolism disturbance and subacute inflammation within the microenvironment of follicular fluid between Chinese polycystic ovarian syndrome ( PCOS ) patients and the controls . METHODS:Serum lipid indexes , steroid hormone levels , and inflammatory cell counts were analyzed .The inflammatory cytokine and apolipoprotein levels were detected in the serum and follicular fluid .The mRNA expression of apolipoproteins and cytokines in the granulose cells was determined by real-time PCR.RESULTS: PCOS patients showed typical obesity accompanied with hyperlipidemia and hyperandrogenemia .Significantly elevated inflammatory cell number and cytokine lev-els were detected in both serum and follicular fluid .The mRNA expression levels of the inflammatory cytokines and apoli-poproteins in the granulose cells from the PCOS patients were higher than those from the controls .CONCLUSION:Elevat-ed apolipoproteins reflect systematic hyperlipidemia in the follicular fluid .Serum lipids and cytokines penetrate follicle-ser-um barrier and get into follicle fluid .Meanwhile , increased intake of apolipoproteins or elevated synthesis of cytokines ( IL-1, IL-6 and TNF-α) by granulose cells could also be crucial to stabilize microenvironment of follicular fluid .Oocyte and subsequent embryos are sensitive to the originaal follicular environment .The lipid metabolism disturbance , inflammatory cell infiltration and hyperandrogenemia may possibly disturb oocyte developmental potential .
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Objective: To investigate the effect of the presence or absence of corpus luteum on hormonal composition of follicular fluid (FF) from different sized follicles and their relationship to serum concentrations in dairy cows. Methods: Ovaries were collected from 30 clinically healthy adult female cows (Holstein Friesian) 4-7 years of age with clinically normal reproductive tracts after slaughtering. Blood samples were collected from the jugular vein before slaughter from each cow. The stage of the cycle in the cows was determined postmortem. The ovaries collected from per cow were classified with corpus luteum (CL
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OBJECTIVE@#To investigate the effect of the presence or absence of corpus luteum on hormonal composition of follicular fluid (FF) from different sized follicles and their relationship to serum concentrations in dairy cows.@*METHODS@#Ovaries were collected from 30 clinically healthy adult female cows (Holstein Friesian) 4-7 years of age with clinically normal reproductive tracts after slaughtering. Blood samples were collected from the jugular vein before slaughter from each cow. The stage of the cycle in the cows was determined postmortem. The ovaries collected from per cow were classified with corpus luteum (CL(+)) and without corpus luteum (CL(-)). FF was aspirated from small (3-5 mm), medium (6-9 mm), and large (10-20 mm) follicles in CL(+) and CL(-) ovaries. Serum and FF samples were analyzed for estradiol-17β, progesterone, testosterone, T3 and T4 concentrations.@*RESULTS@#Results demonstrated that the FF concentrations of estradiol-17β, progesterone and testosterone in different sized follicles categories (small, medium and large follicles in CL(+) and CL(-) ovaries) were significantly higher (P≤0.05) when compared with the serum. The FF concentrations of estradiol-17β and testosterone in same follicle size categories in CL(+) and CL(-) ovaries were also significant (P<0.05). Indeed, concentrations of these hormones in the CL(-) ovaries were higher than those of the CL(+) ovaries. However, there was a statistically significant difference between medium and large follicles for progesterone concentration in CL(+) and CL(-) ovaries (P<0.05). There was a significant correlation between concentration of hormones in serum and FF with increased follicular diameter.@*CONCLUSIONS@#These results indicated that the levels of hormonal composition in the FF were related to follicular size and interestingly to the presence or absence of a corpus luteum. Indeed, the corpus luteum locally affects neighboring follicular compositions during the luteal phase of the estrous cycle in dairy cows.
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Objetivo: Relacionar los niveles de hormonas esteroideas foliculares con el ciclo de estimulación ovárica y sus resultados globales. Métodos: Se incluyeron pacientes < 38 años, con esterilidad de causa masculina, tubárica o desconocida, que recibieron un protocolo largo con agonistas de GnRH y rFSH. Se recogieron las muestras de la primera y segunda aspiración folicular de cada ovario y se realizó un quimioinmunoanálisis de estradiol, progesterona, testosterona y DHEAS. Resultados: Se obtuvieron cifras menores de DHEAS folicular en las pacientes con más días de frenado con agonistas de GnRH (p=0,0003). Cuantos más días de rFSH administrados, mayores fueron los niveles de testosterona y DHEAS folicular (p=0,03; p=0,03). En los resultados globales del ciclo, se obtuvo una correlación negativa entre las cifras de testosterona folicular y el número de complejos puncionados (r= -0,360; p=0,002) y entre la testosterona folicular y el número de embriones de calidad D (r= -0,233; p=0,047). El número de ovocitos maduros fue menor en pacientes con mayores niveles de testosterona folicular (p=0,008). La progesterona folicular fue superior en ovocitos de buena calidad frente a los de calidad no destacable (p=0,006) y muy mala calidad (p=0,04). Conclusiones: Las cifras altas de testosterona folicular se correlacionaron con menor número de complejos puncionados, ovocitos maduros y embriones de calidad D. La buena calidad ovocitaria se asoció a niveles de progesterona folicular superiores.
Objective: To relate the levels of follicular steroid hormones with the ovarian stimulation cycle and its overall results. Method: It was included patients < 38 years old with sterility of male, tubaric or unknown origin who underwent a long protocol with GnRH agonists and rFSH. Samples were obtained from the first and second follicular aspiration of each ovary. A chemiluminescent immunoassay of estradiol, progesterone, testosterone and DHEAS was performed. Results: Figures of follicular DHEAS decreased as the days of treatment with GnRH agonists increased (p=0.0003) and levels of follicular testosterone and DHEAS increased along with the days of treatment with rFSH (p=0.03, p=0.03). In regard to the outcomes of the overall cycle it was found a negative correlation between follicular testosterone levels and the number of punctured complexes (r= -0.360; p=0.002) and between follicular testosterone and the number of D quality embryos (r= -0.233; p=0.047). The number of mature oocytes was lower in patients with higher levels of follicular testosterone (p=0.008). Follicular progesterone was higher in good quality oocytes as compared to those of no remarkable quality (p=0.006) and very poor quality (p=0.04). Conclusions: High levels of follicular testosterone were correlated with a fewer number of punctured complexes, mature oocytes and D quality embryos. Good oocyte quality was associated with higher follicular progesterone levels.
Subject(s)
Humans , Adult , Female , Gonadal Steroid Hormones/analysis , Follicular Fluid/chemistry , Ovulation Induction , Androgens/analysis , Estradiol/analysis , Ovarian Follicle , Prospective Studies , Progesterone/analysisABSTRACT
This prospective study investigated the relationship between anti-Mullerian hormone (AMH) level in the follicular fluid (FF) and the quality of the oocyte and embryo. A total of 65 FF samples from 54 women were included in this study. FF was collected from the largest preovulatory follicle sized> or =20 mm of mean diameter from each ovary. Samples were divided into 3 groups according to the FF AMH levels: below the 33th percentile (low group, FF AMH3.6 ng/mL, n=22). The quality of the ensuing oocytes and embryos was evaluated by fertilization rate and embryo score. FF AMH levels correlated positively with the matched embryo score on day 3 after fertilization (r=0.331, P=0.015). The normal fertilization rate was significantly lower in the low group than in the intermediate group (61.9% vs. 95.5% vs. 77.3%, respectively, P=0.028). Our results suggest that the FF AMH level could be a predictor of the ensuing oocyte and embryo quality.
Subject(s)
Adult , Female , Humans , Anti-Mullerian Hormone/analysis , Embryo, Mammalian/cytology , Fertilization in Vitro , Follicular Fluid/metabolism , Oocytes/cytology , Prospective StudiesABSTRACT
OBJECTIVE: To investigate the association of individual follicular fluid (FF) leptin and adiponectin levels with the quality of the corresponding oocyte and embryo. METHODS: We prospectively enrolled 67 women who underwent controlled ovarian hyperstimulation with 89 FF samples. FF and the corresponding oocyte was obtained from a single dominant preovulatory follicle at the time of oocyte retrieval. Concentrations of leptin and adiponectin were measured by enzyme-linked immunosorbent assay in an individual follicle. The oocyte quality, fertilization rate, and corresponding embryo development were assessed. RESULTS: The FF level of leptin was significantly associated with body mass index (r=0.334, p<0.01). The FF adiponectin level was significantly higher in the normal fertilization group than the abnormal fertilization group (p=0.009) in the non-obese women. A lower FF leptin level was associated with a trend toward mature oocytes, normal fertilization, and good embryo quality, although these relationships were not statistically significant. The leptin:adiponectin ratio of FF did not differ significantly according to oocyte and embryo quality. The quality of the oocyte and embryo was not associated with the FF leptin level tertile. However, the normal fertilization rate was positively associated with FF adiponectin level tertile. There was a trend towards improved oocytes and normal fertilization rates with the lowest tertile of the FF leptin:adiponectin ratio, but this difference was not statistically significant. CONCLUSION: Our results suggest that a high FF adiponectin concentration could be a predictor of normal fertilization. However, the FF leptin concentration and leptin:adiponectin ratio is not significantly related to oocyte maturity and corresponding embryo development.
Subject(s)
Female , Humans , Pregnancy , Adipokines , Adiponectin , Body Mass Index , Embryonic Development , Embryonic Structures , Enzyme-Linked Immunosorbent Assay , Fertilization , Follicular Fluid , Leptin , Oocyte Retrieval , Oocytes , Prospective Studies , SpermatozoaABSTRACT
In the present study, ovarian follicular fluid concentrations of trace elements and biochemical metabolites in relation to follicular size were investigated in dairy cows. Ovaries were recovered from 40 female adult Holstein Friesian cows 5–7 years of age with clinically normal reproductive tracts after slaughtering. The stage of the cycle in the cows slaughtered was diestrus determined post mortem. Visible follicles on the surface of the ovaries were classified, based on their diameter, into (i) small (3-5 mm), (ii) medium (6-9 mm) and (iii) large (10-20 mm) categories. Follicular fluid samples were analyzed for elements (iron, iodine, copper, manganese, zinc, cobalt, molybdenum and selenium) and biochemical metabolites (glucose, cholesterol, triglyceride, total protein, albumin, globulin, urea and creatinine). Results showed that concentrations of trace elements were different between follicles sized categories. Differences in follicular fluid concentrations of iodine and manganese between follicles sized categories were significant (p ≤ 0.05). In addition, differences in follicular fluid concentration of glucose and cholesterol between follicles sized categories (Small, Medium and large follicles) were significant (p ≤ 0.05). These results of the present study suggest that the levels of trace elements and biochemical metabolites composition in the follicular fluid were related to follicular size in dairy cows.
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Follicular fluid (FF) is an avascular compartment within the mammalian ovary separated from the perifollicular stroma by the follicular wall that constitutes a blood-follicle barrier. FF plays a major role in the autocrine and paracrine regulation and also in the physiological, biochemical and metabolic aspects of the nuclear and cytoplasmic maturation of the oocyte and the process of ovulation. This fluid is also composed of locally produced substances within the follicle, which are related to the metabolic activity of follicular cells, and it is also in part an exudate of serum. The ovarian FF provides suitable microenvironment for the development, growth and maturation of the oocyte and is vital for the maintenance of fertility in the female. FF protects the oocyte from factors that induce premature resumption of meiosis, guards the oocyte from proteolytic attack, facilitates its extrusion during ovulation, and enhances sperm attraction, motility, and the acrosome reaction.