ABSTRACT
To examine the effects of clomiphene citrate (CC) on follicular fluid metabolites and related metabolic pathways in rats with polycystic ovary syndrome (PCOS) using non-targeted metabolomics and determine how CC treats ovulation disorder in PCOS. The Sprague Dawley rats were randomly divided into control, model, and CC groups. A PCOS model was established with letrozole. Body weight, ovarian weight, estrus cycles, serum hormone levels, and ovary histopathology of the rats were collected for further evaluation. Moreover, through ultra-performance liquid chromatography-mass spectrometry, the study of follicular fluid metabolites revealed the mechanism of action of CC. CC reduced ovarian weight and regulated estrous cycles and serum hormone levels in PCOS rats but did not affect their body weight. Moreover, the metabolomic results showed that CC adjusted 153 metabolites, among which 16 cross metabolites like testosterone, androstenedione, 17α-hydroxyprogesterone, and cholic acid were considered as potential biomarkers for CC to improve ovulation disorders in PCOS rats. Kyoto Encyclopedia of Genes and Genomes pathway enrichment also showed that the CC group mainly engaged in tryptophan metabolism and steroid hormone biosynthesis. CC can improve ovulation disorders in rats, and its mechanism is related to the regulation of the secretion of serum hormone and follicular fluid metabolites and the amelioration of multi-metabolic pathways.
Subject(s)
Polycystic Ovary Syndrome , Female , Humans , Rats , Animals , Polycystic Ovary Syndrome/metabolism , Fertility Agents, Female/pharmacology , Follicular Fluid/metabolism , Ovulation Induction/methods , Rats, Sprague-Dawley , Clomiphene/pharmacology , Ovulation , Hormones/pharmacology , Body WeightABSTRACT
INTRODUCTION: Psychiatric and cognitive impairment has been observed in premenopausal women with a hormonal disorder called polycystic ovary syndrome (PCOS). This study aimed to explore the possibility of combining pharmacological agents: Carvedilol and Clomiphene citrate, with antiestrogenic, antioxidant and anti-inflammatory properties in letrozole-induced PCOS rats. METHODS: PCOS was induced in rats by the administration of letrozole (1 mg/kg) daily for 21 days. They were subsequently divided into four groups, each receiving either the vehicle or Clomiphene citrate (1 mg/kg) or Carvedilol or a combination of Clomiphene citrate and Carvedilol, respectively from days 22-36. Neurobehavioral studies were conducted on day 35 (Elevated plus maze and Y maze) and day 36 (Novel object recognition). The serum levels of the antioxidants Superoxide dismutase, Catalase, Interleukin 1B (IL-1B), and the gene expression of nuclear factor-erythroid factor 2-related factor 2 (Nrf2), Nuclear Factor k-Beta (NFKB), and acetylcholine esterase in the frontal brain homogenate was determined. RESULT: Both Carvedilol and the combination therapy reversed the anxiety-like behavior, while Clomiphene citrate and the combination therapy ameliorated the spatial and non-spatial memory impairment observed in PCOS rats. Carvedilol, Clomiphene citrate, and the combination therapy increased the serum concentration of SOD and Catalase and decreased the serum concentration of IL-1B. The combination therapy up-regulated the NRF-2, NFKB, and acetylcholine esterase gene expression. CONCLUSION: Study showed that the combination of carvedilol and clomiphene citrate has anxiolytic potential and improved cognitive functions in PCOS rats. This might have been achieved by carvedilol and clomiphene citrate's ability to modulate the cholinergic system and the Nrf2 pathway while downregulating the NFκB signaling pathway.
Subject(s)
Infertility, Female , Polycystic Ovary Syndrome , Animals , Female , Humans , Rats , Acetylcholine , Carvedilol/pharmacology , Carvedilol/therapeutic use , Catalase , Clomiphene/pharmacology , Clomiphene/therapeutic use , Esterases , Fertility Agents, Female/pharmacology , Fertility Agents, Female/therapeutic use , Infertility, Female/drug therapy , Letrozole/pharmacology , NF-E2-Related Factor 2 , Ovulation Induction , Phenotype , Polycystic Ovary Syndrome/metabolism , NF-kappa B/drug effects , NF-kappa B/metabolismABSTRACT
The present study examined the long-term effects of suppressing puberty with a GnRH agonist on reproductive physiology and behavior in female rats. We have recently reported that administration of the GnRH agonist leuprolide acetate (25 µg/kg) daily between postnatal day (PD) 25-50 delayed puberty and disrupted the development of copulatory behavior and sexual motivation in male rats. However, pilot data from our lab suggest that this low dose of leuprolide acetate (25 µg/kg) was not high enough to significantly delay puberty in female rats. Therefore, we injected female Long-Evans rats with leuprolide acetate at a higher dose (50 µg/kg) or 0.9% sterile saline, daily , starting on PD 25 and ending on PD 50. Vaginal opening was monitored daily starting on PD 30 for signs of pubertal onset and first estrous cycle. In addition, we measured estrous cyclicity starting approximately 2 weeks after the last injection of leuprolide (â¼PD 64). Immediately after monitoring estrous cyclicity, the female rats were mated on their first day in behavioral estrus using the partner-preference paradigm, with and without physical contact (PD 95-110). We found that this dose of leuprolide (50 µg/kg) significantly delayed puberty; however, neither estrous cyclicity nor sexual motivation was significantly affected by periadolescent exposure to leuprolide. Together with our findings in male rats, these results add to our understanding of the developmental effects of chemically suppressing puberty in rats.
Subject(s)
Estrous Cycle , Fertility Agents, Female , Leuprolide , Sexual Behavior, Animal , Sexual Maturation , Animals , Estrous Cycle/drug effects , Estrous Cycle/physiology , Estrus , Female , Fertility Agents, Female/pharmacology , Gonadotropin-Releasing Hormone/agonists , Leuprolide/pharmacology , Models, Animal , Periodicity , Rats , Rats, Long-Evans , Sexual Behavior, Animal/drug effects , Sexual Behavior, Animal/physiology , Sexual Maturation/drug effects , Sexual Maturation/physiologyABSTRACT
RESEARCH QUESTION: Polycystic ovary syndrome (PCOS) alters oocyte developmental competence and so the treatment of PCOS patients with assisted reproductive techniques is a major challenge for an infertility specialist. Does ovulation induction therapy with clomiphene citrate, metformin and flutamide affect abnormal PCOS follicle gene expression, its quality, and nuclear and cytoplasmic maturation in the oocyte matured in vitro? DESIGN: Fifty NMRI mice (7-8 weeks old) were randomly divided into five groups, including non-PCOS, PCOS and PCOS groups treated with clomiphene citrate (18 mg/kg body weight for 2 days), metformin (50 mg/100 g body weight for 30 days) and flutamide (10 mg/kg body weight injection for 15 days). The oocytes were collected for quantitative real-time polymerase chain reaction analysis (the evaluation of genes associated with oocyte maturation), transmission electron microscopy (the evaluation of cytoplasmic maturation) and in-vitro maturation (IVM) and IVF outcomes. RESULTS: The dysregulated expression of some genes of insulin-like growth factor (IGF) families involved in oocyte competence and steroid metabolism (e.g. Cyp19a1 and Cyp11a1), transforming growth factor beta (TGF-ß) family (e.g. Tgfb1, Gdf9, Bmp15, Inhbb and Bmpr1a), and oestrogen receptor signalling (e.g. Ncoa3, Pgr) was, to some extent, restored in the PCOS follicles following their with clomiphene citrate, metformin and flutamide. The improved cytoplasmic maturation was observed particularly in the PCOS mice treated with clomiphene citrate. The better IVM-IVF outcomes were also observed in the clomiphene citrate and metformin groups compared with the PCOS group. CONCLUSIONS: This study opens up a new perspective on knowing the effect of ovulation induction therapy on not only nuclear maturation but also ultrastructural characteristics, IVM-IVF outcomes and PCOS oocyte developmental competence.
Subject(s)
Infertility, Female , Metformin , Polycystic Ovary Syndrome , Animals , Body Weight , Clomiphene/pharmacology , Female , Fertility Agents, Female/pharmacology , Fertility Agents, Female/therapeutic use , Flutamide/pharmacology , Humans , Infertility, Female/chemically induced , Metformin/pharmacology , Mice , Oocytes , Ovulation Induction/methods , Polycystic Ovary Syndrome/drug therapyABSTRACT
Clomiphene citrate (CC) and letrozole are ovulatory stimulants that, despite high ovulation rates, achieve low pregnancy rates. This study aimed to investigate the in vitro effects of CC and letrozole, alone or in combination with estradiol, on apoptosis in human cumulus cells. We performed a controlled prospective study using primary cumulus cell cultures from patients undergoing in vitro fertilization (n=22). Alpha-inhibin immunocytochemistry was used to assess cell culture purity and morphology. Cell viability was evaluated by MTT assay, cell cycle status by flow cytometry, and Caspase-3, Bax and SOD-2, and S26 gene expression by qPCR. Cells were treated for 24 hours in 5 conditioned media: CC, CC + estradiol, letrozole, letrozole + estradiol and control. None of the treatments affected cell viability, but letrozole reduced the mean percentage of cells in the S phase compared to control (24.79 versus 21.70, p=0.0014). Clomiphene treatment increased mRNA expression of Bax (4 fold) and SOD-2 (2 fold), which was reversed by co-treatment with estradiol. SOD-2 expression increased in cells treated with letrozole compared to control (4 fold), which was also reversed by estradiol. These findings suggest that clomiphene citrate and letrozole do not significantly affect the viability of human cumulus cells. Still, the expression of genes involved in apoptosis was modulated by these drugs alone and in association with estradiol, suggesting that CC and letrozole may have direct effects on cumulus cells beyond their known mechanisms of action.
Subject(s)
Fertility Agents, Female , Infertility, Female , Cell Cycle , Clomiphene/pharmacology , Clomiphene/therapeutic use , Cumulus Cells , Estradiol/pharmacology , Estradiol/therapeutic use , Female , Fertility Agents, Female/pharmacology , Fertility Agents, Female/therapeutic use , Humans , Infertility, Female/drug therapy , Letrozole/pharmacology , Nitriles/pharmacology , Nitriles/therapeutic use , Ovulation Induction , Pregnancy , Pregnancy Rate , Prospective Studies , Superoxide Dismutase , Triazoles/pharmacology , Triazoles/therapeutic use , bcl-2-Associated X ProteinABSTRACT
RESEARCH QUESTION: What is the influence of ethnicity on the outcome of ovulation induction with clomifene citrate in women with polycystic ovary syndrome (PCOS)? DESIGN: This was a retrospective cohort study. In total, 420 women diagnosed with PCOS who were of Northern European, Mediterranean, African, South-East Asian or South American descent, and who started ovulation induction treatment with clomifene citrate, were included. All women were treated with clomifene citrate according to a standardized treatment regimen. The minimal effective dose of clomifene citrate and prevalence of clomifene resistance (CRA) were assessed, and the chance of becoming ovulatory was predicted. RESULTS: Differences were observed in body mass index (Pâ¯=â¯0.008), waist circumference (P = 0.036) and serum LH, insulin and androgen concentrations (all P < 0.001) in women of different ethnicities with PCOS. Compared with women of Northern European descent, the minimal effective dose of clomifene citrate in women of other ethnic groups was not significantly different. The prevalence of CRA (P = 0.574) was similar in all ethnic groups A similar chance of ovulation (P = 0.504) was predicted for the different ethnic groups. CONCLUSIONS: This is the first study aiming to link ethnicity to ovulation induction outcome in PCOS. Although women of different ethnicities who have PCOS exhibit a variation in phenotypic expression, there do not appear to be differences in the prevalence of clomifene-resistant anovulation or the minimal effective dose of clomifene citrate. Furthermore, a prediction model revealed no significant differences in the predicted chance of ovulation. A larger cohort is needed to validate these findings.
Subject(s)
Infertility, Female , Metformin , Polycystic Ovary Syndrome , Citrates/therapeutic use , Clomiphene/therapeutic use , Ethnicity , Female , Fertility Agents, Female/pharmacology , Fertility Agents, Female/therapeutic use , Humans , Infertility, Female/therapy , Male , Metformin/therapeutic use , Ovulation Induction , Polycystic Ovary Syndrome/drug therapy , Retrospective StudiesABSTRACT
Premature ovarian failure (POF) is a leading cause of women's infertility without effective treatment. Here we show that intravenous injection of Ab4B19, an agonistic antibody for the BDNF receptor TrkB, penetrates into ovarian follicles, activates TrkB signaling, and promotes ovary development. In both natural aging and cyclophosphamide-induced POF models, treatment with Ab4B19 completely reverses the reduction of pre-antral and antral follicles, and normalizes gonadal hormone. Ab4B19 also attenuates gonadotoxicity and inhibits apoptosis in cyclophosphamide-induced POF ovaries. Further, treatment with Ab4B19, but not BDNF, restores the number and quality of oocytes and enhances fertility. In human, BDNF levels are high in granulosa cells and TrkB levels increase in oocytes as they mature. Moreover, BDNF expression is down-regulated in follicles of aged women, and Ab4B19 activates TrkB signaling in human ovary tissue ex vivo. These results identify TrkB as a potential target for POF with differentiated mechanisms, and confirms superiority of TrkB activating antibody over BDNF as therapeutic agents.
Subject(s)
Fertility Agents, Female/pharmacology , Membrane Glycoproteins/agonists , Ovary/drug effects , Primary Ovarian Insufficiency/drug therapy , Receptor, trkB/agonists , Adult , Aging/physiology , Animals , Apoptosis/drug effects , Brain-Derived Neurotrophic Factor/agonists , Brain-Derived Neurotrophic Factor/metabolism , Cell Line, Tumor , Cyclophosphamide/toxicity , Disease Models, Animal , Female , Fertility/drug effects , Fertility Agents, Female/therapeutic use , Humans , Male , Membrane Glycoproteins/metabolism , Mice , Middle Aged , Organ Culture Techniques , Ovary/pathology , Ovary/physiopathology , Primary Ovarian Insufficiency/chemically induced , Primary Ovarian Insufficiency/pathology , Primary Ovarian Insufficiency/physiopathology , Receptor, trkB/metabolism , Young AdultABSTRACT
BACKGROUND: Ovarian tissue cryopreservation and transplantation (OTCTP) is currently the main option available to preserve fertility in prepubertal patients undergoing aggressive cancer therapy treatments. However, a major limitation of OTCTP is follicle loss after transplantation. The mouse is a model of choice for studying ovarian function and follicle development after ovarian tissue grafting in vivo. In these mouse models, ovarian tissue or ovaries can be transplanted to different sites. Our aim was to evaluate a new alternative to heterotopic transplantation models that could be useful to test pharmaceutical improvement for ovarian grafts after OTCTP. METHODS: Slow frozen murine whole ovaries were transplanted into the mouse ears (between the external ear skin layer and the cartilage). Ovarian transplants were recovered after 3, 14 or 21 days. Grafts were analyzed by immunohistochemistry and follicle density analyses were performed. RESULTS: An increase of ovarian vascularization (CD31 and Dextran-FITC positive staining), as well as cellular proliferation (Ki67 staining) were observed 3 weeks after transplantation in comparison to 3 days. Fibrosis density, evaluated after Van Gieson staining, decreased 3 weeks after transplantation. Furthermore, transplantation of cryopreserved ovaries into ovariectomized mice favored follicle activation compared to transplantation into non-ovariectomized mice. CONCLUSION: The present study indicates that surgical tissue insertion in the highly vascularized murine ear is an effective model for ovarian grafting. This model could be helpful in research to test pharmaceutical strategies to improve the function and survival of cryopreserved and transplanted ovarian tissue.
Subject(s)
Drug Evaluation, Preclinical/methods , Fertility Agents, Female/therapeutic use , Fertility Preservation/methods , Ovary/transplantation , Transplantation, Heterotopic/methods , Animals , Cell Proliferation/drug effects , Combined Modality Therapy , Female , Fertility Agents, Female/pharmacology , Graft Survival/drug effects , Hormone Replacement Therapy/methods , Mice , Mice, Inbred BALB C , Mice, SCID , Models, BiologicalABSTRACT
BACKGROUND: Adenomyosis is a chronic gynecological disease characterized by invasion of the uterine endometrium into the muscle layer. In assisted reproductive technology (ART), gonadotropin-releasing hormone agonist (GnRHa) is often used to improve pregnancy rates in patients with adenomyosis, but the underlying mechanisms are poorly understood. METHODS: Eutopic endometrial specimens were collected from patients with adenomyosis before and after GnRHa treatment in the midsecretory phase. RNA sequencing (RNA-Seq) of these specimens was performed for transcriptome analysis. The differentially expressed genes (DEGs) of interest were confirmed by real-time PCR and immunohistochemistry. RESULTS: A total of 132 DEGs were identified in the endometrium of patients with adenomyosis after GnRHa treatment compared with the control group. Bioinformatics analysis predicted that immune system-associated signal transduction changed significantly after GnRHa treatment. Chemokine (C-C motif) ligand 21 (CCL21) was found to be highly expressed in the eutopic endometrium after GnRHa treatment, which may be involved in the improvement of endometrial receptivity in adenomyosis. CONCLUSION: This study suggests that molecular regulation related to immune system-associated signal transduction is an important mechanism of GnRHa treatment in adenomyosis. Immunoreactive CCL21 is thought to regulate inflammatory events and participate in endometrial receptivity in adenomyosis.
Subject(s)
Adenomyosis/genetics , Endometrium/drug effects , Fertility Agents, Female/pharmacology , Transcriptome/drug effects , Adenomyosis/drug therapy , Adenomyosis/metabolism , Adenomyosis/pathology , Adult , Animals , Cohort Studies , Embryo Transfer/methods , Endometrium/metabolism , Endometrium/pathology , Female , Fertility Agents, Female/therapeutic use , Gene Expression Profiling , Gene Expression Regulation/drug effects , Gonadotropin-Releasing Hormone/agonists , Humans , Infertility, Female/etiology , Infertility, Female/genetics , Infertility, Female/metabolism , Infertility, Female/therapy , Mice , Mice, Inbred ICR , PregnancyABSTRACT
BACKGROUND: Polycystic ovary syndrome (PCOS) is a metabolic disease that causes infertility due to anovulation in women in reproductive age. It is known that clomiphene citrate (CC) and tamoxifen citrate (TMX) induce ovulation in women with PCOS. In this study, we aimed to investigate the effects of CC and TMX on the autophagy pathway in PCOS. METHODS AND RESULTS: Experimental PCOS model was induced by letrozole (1 mg/kg) in rats by gavage for 21 days. After the last letrozole administration, rats were treated TMX (1 mg/kg) or CC (1 mg/kg) for 5 days. At the end of the experimental procedures, rats in all groups were sacrificed and ovarian tissues were removed. It was observed that mRNA and protein expressions of LC3-II were significantly higher in TMX and CC groups than control and PCOS groups (p < 0.05), while mRNA and protein expressions of mTOR in TMX and CC groups were found significantly lower than control and PCOS groups (p < 0.05). CONCLUSIONS: In conclusion, present study suggests that TMX and CC induce autophagy in ovaries with PCOS. Autophagy is a promising target for understanding pathophysiology of this disease and for developing more effective and safe new protocols for the treatment of PCOS-related anovulation.
Subject(s)
Infertility, Female , Polycystic Ovary Syndrome , Animals , Autophagy , Clomiphene/pharmacology , Female , Fertility Agents, Female/pharmacology , Humans , Infertility, Female/etiology , Microtubule-Associated Proteins , Ovulation Induction/methods , Polycystic Ovary Syndrome/chemically induced , Polycystic Ovary Syndrome/drug therapy , Polycystic Ovary Syndrome/genetics , Rats , TOR Serine-Threonine Kinases/genetics , Tamoxifen/pharmacologyABSTRACT
Objective: This prospective randomized controlled trial compared the reproductive outcomes of frozen embryo transfer (FET) with hormone replacement treatment (HRT) with or without gonadotropin-releasing hormone agonist (GnRHa) pretreatment. Methods: A total of 133 patients scheduled for HRT-FET mainly because of tubal and/or male factors who received two high-quality cleavage-stage embryos were enrolled at two participating centers. The GnRHa group (n = 65) received GnRHa pretreatment, while the control group (n = 68) did not. Analysis was based on the intention-to-treat (ITT) principle. Results: Among the 133 participants, 130 (97.7%) underwent embryo transfer and 127 (95.5%) completed the protocol. The clinical pregnancy rate according to ITT did not differ between the GnRHa and control groups [39/65 (60.0%) vs. 41/68 (60.3%), p = 0.887]. The implantation rate (47.6% vs. 45.3%, p = 0.713), early pregnancy loss rate (5.1% vs. 19.5%, p = 0.09), and live birth rate (49.2% vs. 50.0%, p = 0.920) were also comparable between groups. Conclusion: Pretreatment with GnRHa does not improve the reproductive outcomes for women receiving HRT-FET. Clinical Trial Registration: The study was registered with the Chinese Clinical Trial Registry (ChiCTR-IOR-17014170; http://www.chictr.org.cn).
Subject(s)
Embryo Transfer/methods , Endometrium/drug effects , Fertility Agents, Female/administration & dosage , Gonadotropin-Releasing Hormone/agonists , Hormone Replacement Therapy/methods , Adult , Birth Rate , Blastocyst , Cryopreservation , Drug Administration Schedule , Embryo Implantation/drug effects , Endometrium/pathology , Female , Fertility Agents, Female/pharmacology , Humans , Infant, Newborn , Intention to Treat Analysis , Live Birth , Male , PregnancyABSTRACT
This study was designed to physiologically investigate the fate of stress related infertility conditions to focus on the regulatory response of reproductive potentials in stress-induced female Wistar rats supplemented with clomifene citrate. 42 apparently healthy female Wistar rats weighing about 120-160 g were used in the study. The animals were randomly distributed into 3 groups after acclimatization for 2 weeks. Group 1 served as the control pregnant rats not induced by restraint, mirrored and intruder stressors, group 2 consisted of rats treated with 0.013 mg/g of clomifene citrate drug and exposed to three different stressors while group 3 represented pregnant rats exposed to different stressors but not treated with clomifene citrate. At the end of 3weeks, the rats were euthanized via cervical dislocation. The uterus and ovary organs were carefully isolated, weighed and examined for histological changes. The reproductive capacities studied were gestation period, mean pup weight, litter size and survival rate respectively. Data collected is expressed in Mean±SEM and one way ANOVA statistics was used for comparison of means while Fisher's LSD was employed for post hoc test and the level of significance is determined at p-value < 0.05. Results from our study revealed that restraint and intruder stressors following supplementation with clomifene citrate produced similar stress response in the gestation length, pub-weights, litter size and percentage of survival. Stress of different nature altered the histoarchitecture of the ovary and the uteri of rats exposed to restraint or intruder stressor. Meanwhile, Clomifene citrate administration produced effect on ovulation and pregnancy outcome of stressed pregnant rats and the survival ratio of the offspring.
Subject(s)
Clomiphene/pharmacology , Fertility Agents, Female/pharmacology , Fertility/drug effects , Infertility, Female/drug therapy , Ovary/drug effects , Reproduction/drug effects , Stress, Psychological/complications , Uterus/drug effects , Animals , Disease Models, Animal , Female , Infertility, Female/blood , Infertility, Female/etiology , Infertility, Female/physiopathology , Luteinizing Hormone/blood , Organ Size , Ovary/metabolism , Ovary/physiopathology , Rats, Wistar , Stress, Psychological/physiopathology , Uterus/metabolism , Uterus/physiopathologyABSTRACT
RESEARCH QUESTION: What are the effects on pregnancy outcome in patients with polycystic ovary syndrome (PCOS) in endometrial preparation cycles for vitrified-warmed embryo transfer with or without gonadotrophin releasing hormone (GnRH) agonist pre-treatment? DESIGN: A total of 212 patients with PCOS referred to Royan Institute, Tehran, Iran, between 20 August 2017 to 20 June 2018 were included. The patients were randomly assigned to receive oestradiol after downregulation with GnRH agonist (group A) or without GnRH agonist down-regulation (group B). RESULTS: A total of 188 patients with PCOS completed the trial, 93 patients in group A and 95 patients in group B. Basal oestradiol and LH levels were significantly higher in group B (26.66 versus 41.61, Pâ¯=â¯0.01 and 0.93 versus 5.33, P < 0.0001, respectively). Clinical pregnancy rates were not significantly different in both groups (31.2% versus 33.7%). Similarly, no significant differences were found between groups A and B in miscarriage (9.7% versus 11.6%), implantation (0.58 versus 0.51) and live birth (21.7% versus 22.1%) rates and for medical complications during pregnancy and neonatal anomalies. CONCLUSIONS: Our findings indicate that endometrial preparation for frozen-thawed embryo transfer with and without ovarian suppression by GnRH agonist provides similar results.
Subject(s)
Embryo Transfer/methods , Endometrium/drug effects , Fertility Agents, Female/therapeutic use , Infertility, Female/therapy , Polycystic Ovary Syndrome/therapy , Adult , Embryo Implantation/drug effects , Endometrium/physiology , Female , Fertility Agents, Female/pharmacology , Fertilization in Vitro/methods , Gonadotropin-Releasing Hormone/agonists , Humans , Infant, Newborn , Infertility, Female/complications , Iran , Male , Ovulation Induction/methods , Polycystic Ovary Syndrome/complications , Pregnancy , Pregnancy Outcome , Pregnancy Rate , Sperm Injections, Intracytoplasmic/methods , VitrificationABSTRACT
Objective: To investigate the factors that influence luteal phase short-acting gonadotropin-releasing hormone agonist (GnRH-a) long protocol and GnRH-antagonist (GnRH-ant) protocol on pregnancy outcome and quantify the influence. About the statistical analysis, it is not correct for the number of gravidities. Methods: Infertile patients (n = 4,631) with fresh in-vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) and embryo transfer were divided into GnRH-a long protocol (n =3,104) and GnRH-ant (n =1,527) protocol groups and subgroups G1 (EMT ≤7mm), G2 (7 mm
Subject(s)
Endometrium/pathology , Fertility Agents, Female/therapeutic use , Infertility/drug therapy , Luteal Phase/drug effects , Adult , Birth Rate , China/epidemiology , Endometrium/drug effects , Female , Fertility Agents, Female/pharmacology , Fertilization in Vitro/methods , Gonadotropin-Releasing Hormone/agonists , Gonadotropin-Releasing Hormone/antagonists & inhibitors , Hormone Antagonists/therapeutic use , Humans , Infant, Newborn , Infertility/epidemiology , Infertility/pathology , Infertility/physiopathology , Luteal Phase/physiology , Organ Size/physiology , Ovulation Induction/methods , Pregnancy , Pregnancy Outcome/epidemiology , Pregnancy Rate , Retrospective Studies , Treatment Outcome , Young AdultABSTRACT
This retrospective study compares four different strategies for managing poor ovarian response (POR), namely, conventional stimulation (300 IUs) IVF-fresh embryo transfer (CONVF), mild stimulation (150 IUs) IVF-fresh embryo transfer (MILDF), mild stimulation embryo banking (MILDB), and embryo banking in natural cycles (NATB). In total, 796 POR patients were considered eligible. Statistical analysis revealed a shorter duration of stimulation and a lower required amount of gonadotropins in MILDF compared with CONVF (9.34 ± 1.17 versus 10.37 ± 1.14; 1402 ± 176 versus 3110 ± 343, P < 0.001). Comparing MILDF and MILDB, a higher number of available oocytes and embryos was observed in MILDB (2.36 ± 1.15 versus 6.58 ± 1.11; 1.72 ± 1.02 versus 3.51 ± 0.61, P < 0.001). Moreover, the MILDB presented with a lower number of required oocyte retrievals and a higher number of oocytes per oocyte retrieval compared with NATB (3.90 ± 1.56 versus 7.15 ± 1.80; 1.95 ± 0.74 versus 0.89 ± 0.20, P < 0.001). Data indicate that MILDF is equally efficient and associated with a shorter duration of stimulation and a lower required amount of gonadotropins compared with CONVF. Embryo accumulation may be more efficient compared with a fresh embryo transfer. MILDB may be a more efficient approach compared with NATB. To conclude, embryo accumulation following mild stimulation appears to form the optimal strategy for POR management. More studies are needed to verify these conclusions.
Subject(s)
Fertility Agents, Female/pharmacology , Fertilization in Vitro/methods , Fertilization in Vitro/statistics & numerical data , Ovary/drug effects , Ovary/physiopathology , Clinical Decision-Making , Disease Management , Female , Humans , Infertility, Female/etiology , Infertility, Female/therapy , Ovarian Reserve , Pregnancy , Pregnancy Outcome , Retrospective StudiesABSTRACT
OBJECTIVE: To investigate whether Zichong granules (, ZCKL), a very effective herbal formula for treating infertility, have an impact on the differentiation of ovarian granulosa cells from human embryonic stem cells (hESCs) in vitro, and to explore the cellular mechanisms of its clinical effects. METHODS: Serum from ZCKL-medicated rats was prepared and used to treat mesoderm cells derived from hESCs for 6 d. Normal rat serum and a set of growth factors were used as negative and positive controls, respectively. RESULTS: ZCKL-medicated rat serum, but not normal rat serum, induced hESCs-derived mesoderm cells to differentiate into functional ovarian granulosa-like cells (OGLCs) in a similar manner to defined growth factors. The induced OGLCs resembled the morphology of native human granulosa cells, expressed granulosa cell-specific markers at both the mRNA and protein levels, produced high levels of estradiol and strongly responded to follicle-stimulating hormone stimulation. Furthermore, mRNA levels of follistatin, mothers against decapentaplegic homolog 8 and bone morphogenetic protein 6 were dynamically changed during the process. CONCLUSION: In the ZCKL treatment of infertility, one mechanism by which ZCKL may act is by influencing ovarian granulosa cell differentiation and development, possibly through the follistatin and BMP/SMAD signaling pathways.
Subject(s)
Cell Differentiation/drug effects , Drugs, Chinese Herbal/pharmacology , Fertility Agents, Female/pharmacology , Granulosa Cells/drug effects , Human Embryonic Stem Cells/drug effects , Infertility, Female/physiopathology , Animals , Cells, Cultured , Female , Granulosa Cells/cytology , Human Embryonic Stem Cells/cytology , Humans , Infertility, Female/drug therapy , Ovary/cytology , Ovary/drug effects , RatsABSTRACT
This study aimed to examine the effects of adding growth hormone (GH) into the in vitro maturation (IVM) culture medium of mouse oocytes on pregnancy outcomes. Cumulus-oocyte complexes (COCs) were cultured in a medium with (GH group, 100 ng/mL) or without (Con group) GH. Thereafter, chromosome morphology, spindle morphology, and mitochondrial function were examined. Embryo development and blastocyst quality after in vitro fertilization were evaluated. After the embryo transfer, the implantation sites and pregnancy outcomes were evaluated. The oocyte maturation rate of the GH group (81.8 ± 9.6%) was compared to that of the Con group (81.3 ± 6.9%, P = 0.928). The proportion of morphologically abnormal spindles in GH-treated oocytes (7.1 ± 0.9%) was significantly lower than control oocytes (13.7 ± 1.3%, P = 0.032), whereas the proportion of morphologically abnormal chromosomes and mitochondrial distribution was similar between the groups. The mitochondrial membrane potential (P < 0.001) and ATP concentration (P < 0.001) in GH-exposed oocytes were higher than those in control oocytes. After fertilization, the blastocyst rate in the GH group (33.8 ± 13.2%) was significantly higher than the Con group (16.2 ± 2.0%, P = 0.003). In addition, inner cell mass (ICM) number (13.91 ± 3.48 vs. 7.00 ± 1.91, P < 0.001), total cell number (47.45 ± 8.39 vs. 37.71 ± 4.15, P = 0.007), and the ratio of ICM/total cell number (29.9 ± 8.2% vs. 18.6 ± 5.0%, P = 0.002) of blastocyst were all higher in GH group. The implantation rate (71.2 ± 1.9% vs. 39.4 ± 16.4%, P < 0.001) and litter size (8.50 ± 3.99 vs. 3.00 ± 1.22, P = 0.018) were significantly higher in the GH group. Although addition of GH into IVM culture medium does not improve oocyte maturation rate, it improves oocyte and embryo quality, which leads to better embryo development and pregnancy outcomes.
Subject(s)
Fertility Agents, Female/pharmacology , Growth Hormone/pharmacology , In Vitro Oocyte Maturation Techniques , Mitochondria/drug effects , Oocytes/drug effects , Animals , Cells, Cultured , Culture Media , Embryo Culture Techniques , Embryo Implantation , Embryo Transfer , Female , Fertilization in Vitro , Litter Size , Mice , Mice, Inbred ICR , Mitochondria/metabolism , Oocytes/metabolism , Pregnancy , Pregnancy Outcome , Recombinant Proteins/pharmacologyABSTRACT
Angiogenesis, where vascular endothelial growth factor (VEGF) is critically involved, is an important factor in endometrial receptivity. Angio-miRNAs form a special class of microRNAs (miRNAs) that target angiogenic genes and regulate angiogenesis. Various studies have shown that ovarian stimulation and exogenous progesterone affect endometrial vascular density. The present research aimed to assess the impact of HMG/HCG and progesterone on miR-16-5p, VEGF protein expression, and angiogenesis in the mouse endometrium during the preimplantation period. Forty adult female mice were divided into four groups: 1) control, 2) ovarian stimulation (HMG and 48 h after HCG IP), 3) progesterone (progesterone IP for 3 days), 4) ovarian stimulation + progesterone (HMG and 48 h after HCG IP) + (progesterone IP for 3 days) groups.The mice were sacrificed 96 h following HCG administration. miR-16-5p, VEGF protein expression, and CD31-positive cell (Endothelial cell) density were specified.The results showed that endothelial cell density,VEGF protein, and miR-16-5p expression increased in all treatment groups, with the maximum increase belonging to the ovarian stimulation + progesterone group. This study provides evidence that ovarian stimulation and progesterone administration enhance endometrial angiogenesis through VEGF protein upregulation. Furthermore, except for miR-16-5p, other miRNAs and molecules appear to be involved in angiogenic pathways, thereby requiring further studies.
Subject(s)
Chorionic Gonadotropin/pharmacology , Endometrium/blood supply , Endometrium/drug effects , Endothelial Cells/drug effects , Fertility Agents, Female/pharmacology , Menotropins/pharmacology , MicroRNAs/metabolism , Neovascularization, Physiologic/drug effects , Ovulation Induction , Progesterone/pharmacology , Vascular Endothelial Growth Factor A/metabolism , Animals , Endometrium/metabolism , Endothelial Cells/metabolism , Female , Mice , MicroRNAs/genetics , Signal TransductionABSTRACT
Sheep is usually a monovular animal; superovulation technology is used to increase the number of offspring per individual and shorten generation intervals. To date, mature FSH superstimulatory treatments have been successfully used in sheep breeding, but much remains unknown about genes, pathways, and biological functions involved in follicular development. Therefore, in this study, we performed transcriptome profiling of small follicles (SFs; 2-2.5 mm), medium follicles (MFs; 3.5-4.5 mm), and large follicles (LFs; > 6 mm) in Mongolian ewes after FSH superstimulation. Furthermore, we identified differentially expressed genes and performed Kyoto Encyclopedia of Genes and Genomes pathway and Gene Ontology enrichment analyses in 3 separate pairwise comparisons. We found that ovarian steroidogenesis was significantly enriched in the SFs versus MFs analysis; the associated genes, cytochrome P450 family 19 (CYP19) and Hydroxy-delta-5-steroid dehydrogenase 3 beta- and steroid delta-isomerase 1 (HSD3B1), were significantly upregulated. Moreover, proline metabolism, glutathione metabolism, and PPAR signaling pathways were significantly enriched in the LFs versus SFs analysis; the associated genes, glutamate-cysteine ligase modifier subunit (GCLM) and cystathionine gamma-lyase (CTH), were significantly upregulated, whereas peroxisome proliferator-activated receptor gamma (PPARγ) was significantly downregulated. In summary, our study provides basic data and possible biological direction to further explore the molecular mechanism of sheep follicular development after FSH superstimulation.
Subject(s)
Follicle Stimulating Hormone/pharmacology , Ovarian Follicle/drug effects , Animals , Aromatase/genetics , Aromatase/metabolism , Cloprostenol/pharmacology , Female , Fertility Agents, Female/pharmacology , Gene Expression Profiling , Gene Expression Regulation/drug effects , Glutamate-Cysteine Ligase/genetics , Glutamate-Cysteine Ligase/metabolism , Gonadotropin-Releasing Hormone/analogs & derivatives , Gonadotropin-Releasing Hormone/pharmacology , Luteolytic Agents/pharmacology , Multienzyme Complexes/genetics , Multienzyme Complexes/metabolism , Ovarian Follicle/growth & development , PPAR gamma/genetics , PPAR gamma/metabolism , Progesterone Reductase/genetics , Progesterone Reductase/metabolism , Reproducibility of Results , Sheep , Steroid Isomerases/genetics , Steroid Isomerases/metabolismABSTRACT
Nerve growth factor (NGF) has been verified to be expressed with higher level in adenomyosis uteri, and its neutralizing antibody has a strong inhibitory influence on inflammation. The present study aimed to explore the effect of anti-NGF on the expression of proteins in uteri of mice-induced adenomyosis and assessed its potential role in improving pregnancy rate. In this study, we established a mouse model of adenomyosis and administrated NGF-neutralizing antibody into mice. The mass spectrometry (MS) analysis of the uteri during the implantation window was performed to explore the essential proteins participating in therapy. Besides, embryos of healthy mice were transferred into the uteri to assess the implantation rate. The results of MS analysis demonstrated that 119 proteins were changed in the adenomyosis group compared with control group, and 126 proteins were differentially expressed in the anti-NGF group compared with the adenomyosis group (fold change > 1.5, P < 0.05. After performing cluster analysis using Mfuzz package, we found that a group of proteins participated in cell-cell adhesion and metabolic processes, which were attenuated in the adenomyosis group, while those were successfully recovered by anti-NGF treatment. Western blotting confirmed that the expression levels of integrin alpha-1 (ITGA1), integrin beta-1 (ITGB1), laminin subunit gamma-1 (LAMC1), and creatine kinase M-type (CKM) were decreased in adenomyosis group, whereas those levels were elevated after anti-NGF treatment. Embryo implantation rate in the adenomyosis group was significantly decreased compared with that in the control group (2.31% vs. 26.15%, P < 0.001) and anti-NGF treatment slightly enhanced the embryo implantation rate in mice with experimentally induced adenomyosis (9.23% vs. 2.31%, P = 0.017). Our results revealed that anti-NGF therapy can improve fertility of mice with experimentally induced adenomyosis, possibly through promoting integrin-related proteins.