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1.
Reprod Biol Endocrinol ; 20(1): 88, 2022 Jun 14.
Artigo em Inglês | MEDLINE | ID: mdl-35701786

RESUMO

BACKGROUND: According to current definitions of Polycystic Ovary Syndrome (PCOS), hyperandrogenism is considered as a key element in the pathogenesis of this common endocrinopathy. However, until now, studies about ovarian androgen profile in women are very rare. Our aim was then to characterise the expression profile of the androgens in follicular fluid of 30 PCOS patients, and compare it to those of 47 Control women and 29 women with only polycystic ovary morphology on ultrasounds (ECHO group). METHODS: A retrospective, single-centre cohort study was performed. The intrafollicular concentrations of the key androgens were assessed and correlated with the intrafollicular levels of some adipokines of interest. Androgens were quantified by mass spectrophotometry combined with ultra-high-performance liquid chromatography, while adipokine concentrations were measured by ELISA assays. RESULTS: In PCOS patients, the intrafollicular concentrations of the androgens synthesised by ovarian theca cells, i.e., 17OH-pregnenolone, dehydroepiandrosterone, Δ4-androstenedione and testosterone, were significantly higher than those of the androgens of adrenal origin, and positively correlated with the main PCOS clinical and biological features, as well as with the adipokines mostly expressed in the follicular fluid of PCOS patients, i.e. resistin, omentin, chemerin and apelin. Conversely, Control women showed the highest levels of 17OH-progesterone, deoxycorticosterone and 11-deoxycortisol. Confirming these results, apelin levels were negatively associated with pregnenolone and deoxycorticosterone concentrations, while visfatin levels, which were higher in the Control group, negatively correlated with the Δ4-androstenedione and testosterone ones. CONCLUSIONS: PCOS is characterised by a selective increase in the intrafollicular levels of the androgens synthesised by theca cells, strengthening the hypothesis that ovarian hyperandrogenism plays a central role in its pathogenesis. Further, the significant correlation between the intrafollicular concentrations of the androgens and most of the adipokines of interest, including apelin, chemerin, resistin and omentin, confirms the existence of a close relationship between these two hormonal systems, which appear deeply involved in ovarian physiology and PCOS physiopathology.


Assuntos
Hiperandrogenismo , Síndrome do Ovário Policístico , Adipocinas , Androgênios/metabolismo , Androstenodiona/metabolismo , Apelina , Estudos de Coortes , Desoxicorticosterona , Feminino , Líquido Folicular/metabolismo , Humanos , Hiperandrogenismo/metabolismo , Síndrome do Ovário Policístico/metabolismo , Pregnenolona , Resistina , Estudos Retrospectivos , Testosterona
2.
Theriogenology ; 188: 116-124, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35689941

RESUMO

The in vitro maturation (IVM) of equine oocytes is still not efficient and does not yield consistent results. The specific requirements of equine oocytes during this process are still largely unknown, which hinders the development of assisted reproductive techniques (ART) in this species. Because the ovarian follicle microenvironment supports oocytes in their acquisition of developmental competence, follicular fluid seems to be a substantial source of bioactive factors that could support the IVM process. Extracellular vesicles (EVs) are cell-secreted molecules in body fluids that are able to deliver molecular signals and transfer genetic information (mRNA, miRNA) between donor and recipient cells. Hence, our hypothesis is that follicular fluid EVs (ffEVs) from small (<20 mm) ovarian follicles can improve the in vitro maturation rate of mare oocytes. To test our hypothesis, equine ovarian follicular fluid was aspirated and ffEVs were isolated by ultracentrifugation, then characterized using nanoparticle tracking analysis and flow cytometry. Additionally, ffEVs were labeled using the ExoGlow-protein EV labeling kit (System Biosciences, Palo Alto, CA). Cumulus-oocyte complexes (COCs) were matured using a one-step method (Method I, continuous culture for 24-38 h) or a two-step method (Method II, initial denudation after 24 h), in the presence (200 µg protein/ml) or absence of ffEVs. The results show the internalization of ffEVs by equine cumulus cells and, for the first time, also by oocytes. The ffEV treatment during two-step culture had a positive effect on the maturation rate of compacted COCs compared to the control group (45.7% and 20.5%, respectively; p < 0.05). No effect of supplementation was observed on the maturation rate during one-step culture. Our results indicate that the supplementation of culture media with EVs isolated from the follicular fluid of small follicles can improve the IVM rate of mare oocytes, suggesting that ffEVs play an important role during this process and may enhance the development of equine ART.


Assuntos
Vesículas Extracelulares , Líquido Folicular , Animais , Células do Cúmulo , Feminino , Cavalos , Técnicas de Maturação in Vitro de Oócitos/métodos , Técnicas de Maturação in Vitro de Oócitos/veterinária , Oócitos , Folículo Ovariano
3.
Sci Rep ; 12(1): 9936, 2022 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-35705692

RESUMO

An increasing number of studies demonstrate that changes in neurotransmitters metabolic levels in follicular fluid are directly related to oocyte maturation, fertilization, the quality of embryo and pregnancy rates. However, the relationship between the intra-follicular neurotransmitters and the function of granulosa cells (GCs), and the outcome of in vitro fertilization-embryo transfer (IVF-ET) is not clear. Human follicular fluid and cumulus GCs were harvested from large follicles obtained from patients undergoing IVF. Neurotransmitters and steroid hormones in follicular fluid were measured through liquid chromatography-tandem mass spectrometry (LC-MS/MS) and high-performance liquid chromatography-mass spectrometry (HPLC-MS/MS). Based on the content of glutamine (Gln) in follicular fluid, the samples were divided into two groups: high Gln level group and low Gln level group. The expression of proliferation-, steroidogenesis- and antioxidant-related genes in GCs was detected by qRT-PCR. In vitro, KGN cells were used to further verify the effects of Gln and NE on GCs function. Primary and secondary outcomes were the number of mature and retrieved oocytes, and the ratio of high-quality embryos, respectively. Gln (46.75 ± 7.74 µg/mL) and norepinephrine (NE, 0.20 ± 0.07 µg/mL) were abundant neurotransmitters in follicular fluid, and exhibited a significantly positive correlation (R = 0.5869, P < 0.005). In high Gln level group, the expression of proliferation, steroidogenesis and antioxidant-related genes in GCs were higher than those in low Gln level group, and the contents of estriol and E2 in follicular fluid were more abundant. Moreover, the concentrations of Gln and NE in follicular fluid showed significantly positive correlation with IDH1 expression in GCs (R = 0.3822, R = 0.4009, P < 0.05). Importantly, a significantly positive correlation was observed between IDH1 expression in GCs and the ratio of higher-quality/cleaved embryos (R = 0.4480, P < 0.05). In vitro studies further demonstrated that Gln and NE played synergistically function in improving GCs proliferation and E2 production by upregulating IDH1 expression. These data demonstrate that Gln and NE in follicular fluid might play significant positive roles in GCs function, and may be potential predictors for selecting optimal quality oocytes and evaluating the quality of embryonic development.


Assuntos
Líquido Folicular , Glutamina , Antioxidantes/metabolismo , Cromatografia Líquida , Feminino , Fertilização In Vitro/métodos , Líquido Folicular/metabolismo , Glutamina/metabolismo , Células da Granulosa/metabolismo , Humanos , Norepinefrina/metabolismo , Oócitos/metabolismo , Gravidez , Espectrometria de Massas em Tandem
4.
Front Endocrinol (Lausanne) ; 13: 865748, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35634503

RESUMO

Polycystic ovarian ovary syndrome (PCOS) is the main cause of ovulatory infertility and a common reproductive endocrine disease of women in reproductive age. In addition, nearly half of PCOS patients are associated with obesity, and their total free fatty acids tend to increase. Arachidonic acid (AA) is a polyunsaturated fatty acid. Oxidation products of AA reacting with various enzymes[cyclooxygenases (COX), lipoxygenases (LOX), cytochrome P450s (CYP)] can change cellular mitochondrial distribution and calcium ion concentration, and increase reactive oxygen species (ROS) production. In this study, we analyzed the follicular fluid fatty acids and found higher levels of C20:4n6 (AA) in PCOS patients than in normal control subjects. Also, to determine whether AA induces oxidative stress (OS) in the human ovarian granulosa tumor cell line (KGN) and affects its function, we treated KGN cells with or without reduced glutathione (GSH) and then stimulated them with AA. The results showed that AA significantly reduced the total antioxidant capacity (TAC) and activity of antioxidant enzymes and increased the malondialdehyde (MDA), ROS and superoxide anion(O2-)levels in KGN cells. In addition, AA was also found to impair the secretory and mitochondrial functions of KGN cells and induce their apoptosis. We further investigated the downstream genes affected by AA in KGN cells and its mechanism of action. We found that AA upregulated the expression of growth differentiation factor 15 (GDF15), which had a protective effect on inflammation and tissue damage. Therefore, we investigated whether AA-induced OS in KGN cells upregulates GDF15 expression as an OS response.Through silencing of GDF15 and supplementation with recombinant GDF15 (rGDF15), we found that GDF15, expressed as an OS response, protected KGN cells against AA-induced OS effects, such as impairment of secretory and mitochondrial functions and apoptosis. Therefore, this study suggested that AA might induce OS in KGN cells and upregulate the expression of GDF15 as a response to OS.


Assuntos
Líquido Folicular , Síndrome do Ovário Policístico , Antioxidantes/metabolismo , Ácido Araquidônico/metabolismo , Linhagem Celular Tumoral , Feminino , Líquido Folicular/metabolismo , Fator 15 de Diferenciação de Crescimento/genética , Fator 15 de Diferenciação de Crescimento/metabolismo , Humanos , Estresse Oxidativo , Síndrome do Ovário Policístico/metabolismo , Espécies Reativas de Oxigênio/metabolismo
5.
Am J Reprod Immunol ; 88(1): e13560, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35506160

RESUMO

OBJECTIVE: To explore the effect of anti-nuclear antibodies (ANAs) on the outcome of in vitro fertilization-embryo transplantation (IVF-ET) and to study the effect of ANAs in follicular fluid (FF) on embryonic development. METHODS: The expression of ANAs in FF of patients treated with IVF-ET and healthy group. The patients were divided into ANAs-positive group and ANA-negative group. The age, duration of infertility, body mass index (BMI), basic follicle stimulating hormone (bFSH), anti-Mullerian hormone (AMH), number of retrieved oocytes, portion of metaphase II oocytes (MII), number of embryos in the cleavage stage, bipronuclear (2PN), number of embryos, number of high-quality embryos and the outcome of IVF-ET were compared between the two groups. In vitro, HTR8/SVneo trophoblast cells were cultivated and divided into ANAs-negative group (control group) and ANAs-positive group (ANAs-positive FF was added to cytotrophoblasts). The ANAs titer in the serum and FF of patients who treated with IVF-ET was detected using ELSIA method. CCK-8 assay and flow cytometry (at 24 h and at 48 h) were used to detect the cell proliferation and apoptosis frequency of the two groups, respectively. RESULTS: Among those who underwent IVF-ET treatment, the number of retrieved oocytes, the number of fertilization and the portion of MII oocytes in the FF-positive group were significantly lower than those in the FF-negative group. Furthermore, the implantation rate and the clinical pregnancy rate were decreased, and early miscarriage rate was increased in the FF-positive patients than those in the FF-negative patients. In vitro, the cytotrophoblasts proliferation activity in the ANAs group was significantly lower than that in the control group. Moreover, the cytotrophoblasts apoptosis rate in the ANAs group was significantly higher than that in the control group. CONCLUSIONS: Our data suggested that ANAs in FF might become an obstacle to embryonic development through promoting trophoblast apoptosis and inhibiting trophoblast proliferation. ANAs in FF might be an unfavorable factor for the outcome of those who undergo IVF-ET treatment.


Assuntos
Anticorpos Antinucleares , Líquido Folicular , Transferência Embrionária , Feminino , Fertilização In Vitro/métodos , Líquido Folicular/metabolismo , Humanos , Oócitos , Gravidez , Fatores de Risco
6.
Sci Total Environ ; 838(Pt 3): 156323, 2022 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-35636536

RESUMO

As follicular fluid constitutes a critical microenvironment for the development of oocytes, investigation of environmental contaminants in follicular fluid may facilitate a better understanding of the influence of environmental exposure on reproductive health. In the present study, we aimed to investigate per- and polyfluoroalkyl substances (PFAS) exposure in women receiving in vitro fertilization-embryo transfer (IVF-ET) treatment, determine the blood-follicle transfer efficiencies (BFTE) of PFAS, and explore potential associations between PFAS exposure and selected IVF-ET outcomes. Our results revealed that n-PFOA was the most abundant PFAS in both serum and follicular fluid (FF) (median = 5.85 and 5.56 ng/mL, respectively), followed by n-PFOS (4.95 and 4.28 ng/mL), 6:2 Cl-PFESA (2.18 and 2.10 ng/mL), PFNA (1.37 and 1.37 ng/mL), PFUdA (0.33 and 0.97 ng/mL), PFDA (0.37 and 0.66 ng/mL), PFHxS (0.42 and 0.39 ng/mL), and PFHpS (0.11 and 0.10 ng/mL). The median BFTE ranged from 0.65 to 0.92 for individual PFAS, indicating a relatively high tendency of PFAS to cross the blood-follicle barrier (BFB). An inverted V-shaped trend was observed between the median BFTE and the number of fluorinated carbon atoms or the log Kow (octanol-water partition coefficient) for individual PFAS, suggesting the influence by physicochemical properties and molecular structures. Although our data did not find any clear pattern in the link between blood or follicular fluid concentrations of PFAS and selected IVF-ET outcomes, our study raises the need for better characterization of exposure to environmental chemicals in follicular fluid together with its potential influence on reproductive health.


Assuntos
Ácidos Alcanossulfônicos , Poluentes Ambientais , Fluorcarbonetos , Transferência Embrionária , Exposição Ambiental , Feminino , Fertilização In Vitro , Fluorcarbonetos/análise , Líquido Folicular/química , Humanos
7.
Cell Commun Signal ; 20(1): 61, 2022 05 09.
Artigo em Inglês | MEDLINE | ID: mdl-35534864

RESUMO

OBJECTIVE: Polycystic ovary syndrome (PCOS) is characterized by follicular dysplasia. An insufficient glycolysis-derived energy supply of granulosa cells (GCs) is an important cause of follicular dysplasia in PCOS. Follicular fluid (FF) exosomal microRNAs (miRNAs) have been proven to regulate the function of GCs. In this study, exosomes extracted from clinical FF samples were used for transcriptome sequencing (RNA-seq) analysis, and a human ovarian granulocyte tumour cell line (KGN cells) was used for in vitro mechanistic studies. METHODS AND RESULTS: In FF exosomal RNA-seq analysis, a decrease in glycolysis-related pathways was identified as an important feature of the PCOS group, and the differentially expressed miR-143-3p and miR-155-5p may be regulatory factors of glycolysis. By determining the effects of miR-143-3p and miR-155-5p on hexokinase (HK) 2, pyruvate kinase muscle isozyme M2 (PKM2), lactate dehydrogenase A (LDHA), pyruvate, lactate and apoptosis in KGN cells, we found that upregulated miR-143-3p expression in exosomes from the PCOS group inhibited glycolysis in KGN cells; knockdown of miR-143-3p significantly alleviated the decrease in glycolysis in KGN cells in PCOS. MiR-155-5p silencing attenuated glycolytic activation in KGN cells; overexpression of miR-155-5p significantly promoted glycolysis in KGN cells in PCOS. In this study, HK2 was found to be the mediator of miR-143-3p and miR-155-5p in FF-derived exosome-mediated regulation of glycolysis in KGN cells. Reduced glycolysis accelerated apoptosis of KGN cells, which mediated follicular dysplasia through ATP, lactate and apoptotic pathways. CONCLUSIONS: In conclusion, these results indicate that miR-143-3p and miR-155-5p in FF-derived exosomes antagonistically regulate glycolytic-mediated follicular dysplasia of GCs in PCOS. Video Abstract.


Assuntos
MicroRNAs , Síndrome do Ovário Policístico , Proliferação de Células , Feminino , Líquido Folicular/metabolismo , Glicólise , Células da Granulosa/metabolismo , Células da Granulosa/patologia , Humanos , Lactatos/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Síndrome do Ovário Policístico/genética , Síndrome do Ovário Policístico/metabolismo , Síndrome do Ovário Policístico/patologia
8.
Reproduction ; 164(1): 1-8, 2022 05 23.
Artigo em Inglês | MEDLINE | ID: mdl-35521903

RESUMO

In Brief: Polycystic ovary syndrome (PCOS) is a common cause of anovulatory infertility in women. This study identified changes in free fatty acids profiles in the follicular fluid that may lead to better diagnosis and management of infertility in PCOS women. Abstract: Polycystic ovary syndrome (PCOS) is a heterogeneous disease characterized by various endocrine/metabolic disorders and impaired reproductive potential. Alterations in oocyte competence are considered potentially causative factors for infertility in PCOS women and analyzing the composition of follicular fluid in these patients may help to identify which changes have the potential to alter oocyte quality. In this study, free fatty acid metabolic signatures in follicular fluid were performed to identify changes that may impact oocyte competence in non-obese PCOS women. Sixty-four non-obese women (32 with PCOS and 32 age- and BMI-matched controls) undergoing in vitro fertilization were recruited. Embryo quality was morphologically assessed. Free fatty acid metabolic profiling in follicular fluid was performed using gas/liquid chromatography-mass spectrometry. Principal component analysis and orthogonal partial least squares-discriminant analysis models were further constructed. Nine free fatty acids and 24 eicosanoids were identified and several eicosanoids synthesized by the cyclooxygenase pathway were significantly elevated in PCOS patients compared to controls. The combination of PGE2, PGF2α, PGJ2, and TXB2 had an area under the curve of 0.867 (0.775-0.960) for PCOS discrimination. Furthermore, follicular fluid levels of PGE2 and PGJ2 were negatively correlated with high-quality embryo rate in PCOS patients (P < 0.05). Metabolomic analysis revealed that follicular fluid lipidomic profiles undergo changes in non-obese PCOS women, which suggests that identifying changes in important metabolic signatures may give us a better understanding of the pathogenesis of PCOS. Furthermore, elevated PGE2 and PGJ2 concentrations may contribute to impaired oocyte competence in non-obese PCOS patients.


Assuntos
Infertilidade Feminina , Síndrome do Ovário Policístico , Dinoprostona/metabolismo , Ácidos Graxos não Esterificados , Feminino , Líquido Folicular/metabolismo , Humanos , Infertilidade Feminina/metabolismo , Oócitos/metabolismo , Síndrome do Ovário Policístico/complicações , Síndrome do Ovário Policístico/metabolismo
9.
Sci Prog ; 105(2): 368504221103782, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35619572

RESUMO

BACKGROUND: The level of vitamin D in follicular fluid (FF) according to the ovarian reserve has never been investigated, and the effect of FF vitamin D on the outcome of assisted reproductive technology (ART) remains controversial. The aim of this study is to evaluate the association between FF vitamin D levels and baseline anti-Müllerian hormone (AMH) / ART outcomes. METHODS: Forty-seven patients who underwent controlled ovarian stimulation at the fertility clinic of an academic tertiary care center were enrolled for a prospective observational study. FF was collected from the first aspirated leading follicle of each ovary and assayed by an enzyme-linked immunosorbent assay. Multivariable linear regression analysis was used to assess the association between baseline AMH and FF vitamin D levels with adjustment for basal FSH and serum vitamin D levels. RESULTS: Both the AMH and serum vitamin D were significant predictors for FF vitamin D. The estimated marginal mean of FF vitamin D level was higher in women with decreased ovarian reserve (DOR) than those with normal ovarian reserve (24.1 ± 2.1 vs. 18.8 ± 1.4 ng/ml, p = 0.048). However, FF vitamin D did not demonstrate any significant associations with cycle outcomes, including fertilization rate and the number and proportion of good embryos at day three. CONCLUSION: We observed significantly higher FF vitamin D levels in women with DOR. However, FF vitamin D did not demonstrate any significant associations with the outcome of ART. A larger prospective study is needed to investigate the effect of FF vitamin D on the clinical pregnancy rate and live birth rate.


Assuntos
Líquido Folicular , Reserva Ovariana , Hormônio Antimülleriano/análise , Feminino , Fertilização In Vitro , Líquido Folicular/química , Humanos , Reserva Ovariana/fisiologia , Projetos Piloto , Gravidez , Vitamina D , Vitaminas
10.
Reprod Biol Endocrinol ; 20(1): 70, 2022 Apr 21.
Artigo em Inglês | MEDLINE | ID: mdl-35448997

RESUMO

BACKGROUND: Nicotinamide (NAM) is an important antioxidant, which is closely related to female fertility, but its role has not been clearly elucidated. The purpose of the present study was to investigate the effects of NAM on follicular development at different stages and the quality of oocytes. METHODS: The concentration of NAM in follicular fluid (FF) of 236 women undergoing in vitro fertilization (IVF) was ascertained by enzyme-linked immunosorbent assay (ELISA), and the correlation between NAM and clinical indexes was analyzed. During the in vitro maturation (IVM) of mice cumulus-oocyte complexes (COCs), different concentrations of NAM were added to check the maturation rate and fertilization rate. The reactive oxygen species (ROS) levels in the oocytes treated with different hydrogen peroxide (H2O2) and NAM were assessed. Immunofluorescence staining was performed to measure the proportion of abnormal spindles. RESULTS: The level of NAM in large follicles was significantly higher than that in small follicles. In mature FF, the NAM concentration was positively correlated with the rates of oocyte maturation and fertilization. Five mM NAM treatment during IVM increased maturation rate and fertilization rate in the oxidative stress model, and significantly reduced the increase of ROS levels induced by H2O2 in mice oocytes. CONCLUSIONS: Higher levels of NAM in FF are associated with larger follicle development. The supplement of 5 mM NAM during IVM may improve mice oocyte quality, reducing damage caused by oxidative stress.


Assuntos
Peróxido de Hidrogênio , Técnicas de Maturação in Vitro de Oócitos , Animais , Feminino , Fertilização In Vitro , Líquido Folicular , Humanos , Peróxido de Hidrogênio/farmacologia , Masculino , Camundongos , Niacinamida/farmacologia , Oócitos , Espécies Reativas de Oxigênio
11.
Medicina (Kaunas) ; 58(4)2022 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-35454338

RESUMO

Background and objectives. Gestational diabetes mellitus is an increasingly diagnosed metabolic disorder during pregnancy with unknown pathological pathways. Taking into account the growing numbers of women who are conceiving after assisted reproductive technologies, they comprise an engaging target group for gestational diabetes mellitus etiopathogenesis research. In terms of metabolism and genetics, as the evidence shows, both unexplained infertility and gestational diabetes mellitus pose challenges for their interpretation due to the complex bodily processes. Materials and Methods. Our study examined the expression of genes (IGF2, GRB10, CRTC2, HMGA2, ESR1, DLK1, SLC6A15, GPT2, PLAGL1) associated with glucose metabolism in unexplained infertility patients who conceived after in vitro fertilization procedure, were diagnosed with GDM and their findings were compared with control population. Results. There were no significant differences in gene expression of endometrium stromal cells between healthy pregnant women and women with gestational diabetes, although the significant downregulation of CRTC2 was observed in the follicular fluid of women with gestational diabetes mellitus. Moreover, expression of HMGA2 and ESR1 was significantly reduced in FF cells when compared to endometrial cells. Conclusions. These findings may indicate about the importance of follicular fluid as an indicator for gestational diabetes and should be explored more by further research.


Assuntos
Diabetes Gestacional , Endométrio , Líquido Folicular , Infertilidade , Diabetes Gestacional/epidemiologia , Diabetes Gestacional/genética , Feminino , Humanos , Infertilidade/complicações , Gravidez , Prognóstico
12.
Dis Markers ; 2022: 6877541, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35465261

RESUMO

Aims: To explore the effect of IR on the metabolism of PCOS by analyzing the changes in FF metabolites in PCOS patients who are undergoing assisted reproductive technology based on the metabonomic platform of ultraperformance gas chromatography coupled to mass spectrometry (GC/MS). Method: Eight PCOS patients with IR (PCOS-IR) and 8 PCOS patients without IR (PCOS-NIR) were enrolled. All patients received controlled ovarian stimulation by using the gonadotropin-releasing hormone (GnRH) antagonist protocol, and the FF of a single dominant follicle was collected on the day of oocyte retrieval. The metabolite profiles of the FF were determined by GC/MS. Key Results. A total of 20 differentially expressed metabolites in FF were identified. Compared with levels in the PCOS-NIR group, stearic acid, palmitic acid, pentadecanoic acid, stigmasterol, citric acid, isocitric acid, thymine, and pyruvic acid in FF were significantly increased in the PCOS-IR group. Lithocholic acid and sinapinic acid in FF decreased significantly. The affected metabolic pathways with potential regulatory roles were identified by KEGG annotation. Conclusion: Compared with the PCOS-NIR group, the PCOS-IR group showed more significant metabolic abnormalities. Implications. These results will help us to understand the pathogenesis of PCOS combined with IR and will provide new clues for studying metabolic disorders associated with PCOS, e.g., IR.


Assuntos
Resistência à Insulina , Síndrome do Ovário Policístico , Feminino , Líquido Folicular/química , Líquido Folicular/metabolismo , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Metabolômica/métodos
13.
Int J Mol Sci ; 23(8)2022 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-35456957

RESUMO

In goats, embryo oocyte competence is affected by follicle size regardless the age of the females. In previous studies we have found differences in blastocyst development between oocytes coming of small (<3 mm) and large follicles (>3 mm) in prepubertal (1-2 months-old) goats. Oocyte competence and Follicular Fluid (FF) composition changes throughout follicle growth. The aim of this study was to analyze Fatty Acids (FAs) composition and metabolomic profiles of FF recovered from small and large follicles of prepubertal goats and follicles of adult goats. FAs were analyzed by chromatography and metabolites by 1H-Nuclear Magnetic Resonance (1H-NMR) Spectrometry. The results showed important differences between adult and prepubertal follicles: (a) the presence of α,ß-glucose in adult and no detection in prepubertal; (b) lactate, -N-(CH3)3 groups and inositol were higher in prepubertal (c) the percentage of Linolenic Acid, Total Saturated Fatty Acids and n-3 PUFAs were higher in adults; and (d) the percentage of Linoleic Acid, total MUFAs, PUFAs, n-6 PUFAs and n-6 PUFAs: n-3 PUFAs ratio were higher in prepubertal goats. Not significant differences were found in follicle size of prepubertal goats, despite the differences in oocyte competence for in vitro embryo production.


Assuntos
Líquido Folicular , Cabras , Animais , Ácidos Graxos/metabolismo , Feminino , Fertilização In Vitro/veterinária , Líquido Folicular/metabolismo , Oócitos/metabolismo , Folículo Ovariano/metabolismo
14.
J Med Invest ; 69(1.2): 112-116, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35466131

RESUMO

Oxidative stress (OS) is a situation that occurs as a result of un-equilibrium between reactive oxygen species (ROS) formation and the antioxidant defence system. 8-hydroxy-2-deoxyguanosine (8-OHdG), an oxidized form of deoxyguanosine, is found in higher levels in aging oocytes. In this cross-sectional study, we aimed to evaluate the effect of 8-OHdG on the intracytoplasmic sperm injection (ICSI). Follicular fluid (FF) samples were taken after removing the oocyte for later analysis of 8-OHdG. The couples participants were categorized according to the cause of subfertility into three groups (female factor, malefactor, and unexplained infertility). Further division according to pregnancy state was done to evaluate the precise role of 8-OHdG on pregnancy state and ICSI outcome. Result : This study showed that the 8-OHdG levels significantly higher in non-pregnant women (p<0.05). Correlation study showed that 8-OHdG level in follicular fluid is negatively correlating with the number of retrieved oocytes, metaphase two oocytes (MII), fertilized oocytes (2PN = two pronuclei), cleaved zygotes, good quality embryos, and the difference was statistically significant (p<0.05). Conclusion: The study revealed that the 8-OHdG level in follicular fluid negatively influences ICSI outcome and it is higher in non-pregnant women. J. Med. Invest. 69 : 112-116, February, 2022.


Assuntos
Líquido Folicular , Injeções de Esperma Intracitoplásmicas , 8-Hidroxi-2'-Desoxiguanosina , Biomarcadores , Estudos Transversais , Feminino , Líquido Folicular/metabolismo , Humanos , Estresse Oxidativo , Gravidez , Taxa de Gravidez
15.
J Med Invest ; 69(1.2): 65-69, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35466148

RESUMO

It has been shown that biotin, a water-soluble vitamin (B7), plays roles in reproductive functions, such as oocyte maturation and embryo development, in experimental animals. On the other hand, little is known about the clinical effects of biotin on human reproduction. In this study, serum and follicular fluid biotin levels were measured in patients who underwent in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI), and their associations with reproductive outcomes were evaluated. As a result, biotin was detected in follicular fluid, as well as serum, and the biotin levels of follicular fluid were found to be positively correlated with those of serum. The biotin levels of serum were higher than those of follicular fluid, suggesting that biotin may be taken up into the follicular fluid from the blood. Although serum and follicular fluid biotin levels tended to be higher in pregnant patients than in non-pregnant patients, these data did not show the significant statistical difference. These findings indicate that biotin does not contribute to the maintenance of oocyte quality, and hence, it does not increase fertilization and pregnancy rates. J. Med. Invest. 69 : 65-69, February, 2022.


Assuntos
Líquido Folicular , Injeções de Esperma Intracitoplásmicas , Biotina , Feminino , Fertilização In Vitro , Humanos , Gravidez , Resultado da Gravidez
16.
Sci Rep ; 12(1): 4359, 2022 03 14.
Artigo em Inglês | MEDLINE | ID: mdl-35288625

RESUMO

Polycystic ovary syndrome (PCOS) is an endocrine disorder that occurs in women of reproductive age. Anovulation caused by abnormal follicular development is still the main characteristic of PCOS patients with infertile. Granulosa cell (GC) is an important part of the follicular microenvironment, the dysfunction of which can affect follicular development. Increasing evidence indicates that exosomal miRNAs derived from the follicular fluid (FF) of patients play critical roles during PCOS. However, which follicular fluid-derived exosomal miRNAs play a pivotal role in controlling granulosa cell function and consequently follicular development remain largely unknown, as does the underlying mechanism. Herein, we showed that miR-143-3p is highly expressed in the follicular fluid exosomes of patients with PCOS and can be delivered into granulosa cells. Furthermore, functional experiments showed that translocated miR-143-3p promoted granulosa cell apoptosis, which is important in follicle development. Mechanistically, BMPR1A was identified as a direct target of miR-143-3p. Overexpression of BMPR1A reversed the effects of exosomal miR-143-3p on GC apoptosis and proliferation by activating the Smad1/5/8 signaling pathway. These results demonstrate that miR-143-3p-containing exosomes derived from PCOS follicular fluid promoted granulosa cell apoptosis by targeting BMPR1A and blocking the Smad1/5/8 signaling pathway. Our findings provide a novel mechanism underlying the roles of exosomal-miRNAs in the follicular fluid of PCOS patients and facilitate the development of therapeutic strategies for PCOS.


Assuntos
MicroRNAs , Síndrome do Ovário Policístico , Apoptose/genética , Receptores de Proteínas Morfogenéticas Ósseas Tipo I/metabolismo , Proliferação de Células/genética , Feminino , Líquido Folicular/metabolismo , Células da Granulosa/metabolismo , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , Síndrome do Ovário Policístico/genética , Síndrome do Ovário Policístico/metabolismo , Microambiente Tumoral
17.
Reprod Biol ; 22(2): 100633, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35278823

RESUMO

The role of adipokines in ovarian-related disorders such as polycystic ovary syndrome (PCOS) has been reported. However, the involvement of Oncostatin M (OSM), a recently identified adipokine, in ovarian function is unknown. Therefore, we investigated the association of the OSM signaling pathway with ovarian functions and PCOS pathogenesis. This case-control study enrolled 30 PCOS and 30 healthy women who underwent the intracytoplasmic sperm injection procedure. OSM and OSM receptor (OSMR) levels were evaluated in the follicular fluid (FF). Moreover, the expression of insulin receptor substrates (IRS1 and IRS2), OSM, OSMR, suppressor of cytokine signaling 3 (SOCS3), and androgen receptor (AR) genes were analyzed in the isolated cumulus cells (CCs). For the in-vitro experiment, the effect of recombinant OSM on the expression of related genes in isolated CCs was analyzed. Follicular concentrations of OSM and OSMR were significantly lower in PCOS (123.91±48.58 pg/ml and 0.93±0.35 ng/ml, respectively) compared to control women (283.53 ± 96.62 pg/ml and 1.45 ± 0.18 ng/ml, respectively; p < 0.001) and were positively correlated with the oocyte maturation (r = 0.611 and r = 0.611, respectively) and fertilization (r = 0.592 and r = 0.627, respectively) rates in the PCOS group. Furthermore, the SOCS3 expression was upregulated about eight times in PCOS patients compared to the controls (p < 0.05). The treatment of cells with recombinant OSM significantly increased SOCS3, OSMR, IRS-1, and -2 expression and decreased AR expression. The decreased levels of OSM and its receptor in PCOS patients, possibly mediated by SOCS3, could negatively affect oocyte maturation and fertilization rates.


Assuntos
Síndrome do Ovário Policístico , Estudos de Casos e Controles , Feminino , Líquido Folicular/metabolismo , Humanos , Oncostatina M/metabolismo , Síndrome do Ovário Policístico/metabolismo , Técnicas de Reprodução Assistida
18.
Reprod Biol Endocrinol ; 20(1): 54, 2022 Mar 21.
Artigo em Inglês | MEDLINE | ID: mdl-35313911

RESUMO

BACKGROUND: Both mild and conventional controlled ovarian stimulation are the frequently used protocols for poor ovarian responders. However, there are some debates about which treatment is better. Moreover, little is known about the follicular physiology after the two ovarian stimulation protocols. This study was intended to investigate the features in granulosa cells and follicular fluid micro-environment after the two different ovarian stimulation protocols in poor responders. METHODS: Granulosa cells RNA were sequenced using Illumina Hiseq technology. Specific differently expressed genes and proteins were verified by real-time quantitative PCR and Western blot analysis. Moreover, hormone and cytokine concentrations in the follicular fluid were measured by electrochemiluminescence immunoassay and enzyme-linked immunoabsorbent assay. The correlation between the results of molecular experiments and the laboratory outcomes were analyzed by Spearman correlation analysis. RESULTS: The differentially expressed genes between the two groups were involved in 4 signaling pathways related to the follicular development; three proteins pertinent to the TGF-ß signaling pathway were expressed differently in granulosa cells between the two, and the constituents in the follicular fluid were also different. Further, a correlation between the TGF-ß signaling pathway and the good-quality embryo was observed. CONCLUSIONS: The present study made a comparison for the first time in the transcriptome of human granulosa cells and the follicular fluid micro-environment between poor responders with the conventional controlled ovarian stimulation or the mild ovarian stimulation, showing that the TGF-ß signaling pathway may correlate with the good-quality of embryos in the mild group, which may be instrumental to the choice of optimal management for IVF patients.


Assuntos
Líquido Folicular/metabolismo , Células da Granulosa/metabolismo , Infertilidade Feminina/genética , Indução da Ovulação/métodos , Transcriptoma , Adulto , Estudos de Casos e Controles , Microambiente Celular/genética , Feminino , Líquido Folicular/química , Perfilação da Expressão Gênica , Células da Granulosa/química , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Infertilidade Feminina/metabolismo , Infertilidade Feminina/fisiopatologia , Recuperação de Oócitos , Reserva Ovariana/genética , Ovulação/genética , Análise de Sequência de DNA , Falha de Tratamento
19.
Fertil Steril ; 117(4): 771-780, 2022 04.
Artigo em Inglês | MEDLINE | ID: mdl-35272846

RESUMO

OBJECTIVE: To study the presence of viral RNA in the follicular fluid, cumulus cells, and endometrial tissue samples in SARS-CoV-2-positive women undergoing assisted reproductive technology (ART). DESIGN: Prospective, single-center, observational study. SETTING: Tertiary hospital. PATIENT(S): A total of 16 patients undergoing transvaginal oocyte retrieval who had a positive SARS-CoV-2 RNA test <48 hours before the procedure. All patients underwent the retrieval between September 2020 and June 2021 and used in vitro fertilization or intracytoplasmic sperm injection. All embryos were vitrified to avoid conception during SARS-CoV-2 infection. INTERVENTION(S): Follicular fluid aspirated during oocyte retrieval, cumulus cells, and endometrial samples were analyzed for SARS-CoV-2 RNA using the RealStar SARS-CoV-2 RT-PCR-Kit1.0. MAIN OUTCOME MEASURE(S): The primary outcome parameter was the detection of viral RNA in the follicular fluid, cumulus cells, and endometrial cells. Fertilization rate, embryo developmental potential, and clinical outcome after frozen embryo transfer were secondary outcome parameters. RESULT(S): Samples from 16 patients were analyzed. Cycle threshold values of <40 were considered positive. All samples were negative for SARS-CoV-2 viral RNA. No inflammatory lesions of the endometrium were identified histologically. Fertilization rate, embryo development, and clinical outcomes after embryo transfer were reassuring. CONCLUSION(S): In women infected with SARS-CoV-2 who underwent ART, viral RNA was undetectable in the follicular fluid, cumulus cells, and endometrium. Caution is warranted in view of the small sample size, and the risk of SARS-CoV-2 affecting the embryo via ART cannot be ruled out. Adequate counseling of women and couples undergoing ART is crucial in parallel with further research on the effect of exposure of the early human embryo to SARS-CoV-2. CLINICAL TRIAL REGISTRATION NUMBER: NCT04425317.


Assuntos
COVID-19 , RNA Viral , COVID-19/diagnóstico , Células do Cúmulo , Feminino , Fertilização In Vitro/efeitos adversos , Líquido Folicular , Humanos , Estudos Prospectivos , RNA Viral/genética , SARS-CoV-2
20.
Environ Pollut ; 304: 119147, 2022 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-35314206

RESUMO

The blood-follicle barrier (BFB) between the blood and follicular fluid (FF) can maintain the microenvironment balance of oocyte. Boron, an exogenous environmental trace element, has been found to possibly play an important role in oocyte maturation. This study aimed to examine the distribution characteristics of boron across the BFB and find the potential effect of boron on FF microenvironment. We analyzed the concentration of boron in paired FF and serum collected from 168 women undergoing in vitro fertilization and embryo transfer in Beijing City and Shandong Province, China. To explore the potential health impact of boron enrichment in oocyte maturation, a global proteomics analysis was conducted to tentatively correlate the protein levels with the boron enrichment. Interestingly, the results showed that the concentration of boron in FF (34.5 ng/mL) was significantly higher than that in serum (22.0 ng/mL), with a median concentration ratio of 1.52. Likewise, the concentrations of boron in FF and serum were positively correlated (r = 0.446), suggesting that boron concentration in serum can represent its concentration in follicular fluid to a large extent.. This is the first time to observe the enrichment of boron in the FF to our knowledge. It is interesting to observe a total of 13 proteins, which mainly belong to immunoglobulin class, were positively correlated with boron concentration in FF. We concluded that boron, as one environmental trace element, was enriched in FF from blood validated by two area in north china, which may be involved in an increased level of immune processes of immunoglobulins.


Assuntos
Líquido Folicular , Oligoelementos , Boro/metabolismo , Feminino , Fertilização In Vitro/métodos , Humanos , Imunidade , Masculino , Oócitos/metabolismo , Oligoelementos/metabolismo
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