Cause Of Premature Ovarian Failure Research Articles

Proanthocyanidins protects 3-NPA-induced ovarian function decline by activating SESTRIN2-NRF2-mediated oxidative stress in mice

Abnormal apoptosis of ovarian cells caused by oxidative stress is an important cause of premature ovarian failure (POF). Previous studies revealed that proanthocyanidins (PCs) are powerful natural antioxidants that can safely prevent oxidative damage in humans. However, the protective effect and mechanism of PCs on ovarian function during the course of POF remain unknown. In this study, female mice were injected with 3-nitropropionic acid (3-NPA) to establish an ovarian oxidative stress model; at the same time, the mice were treated with PC via gavage. Thereafter, the expression of various apoptosis genes, hormones, and related molecules was assessed. Compared with those in the control group, the ovarian index, follicle count at all levels, expression of MVH, PCNA and BCL2, and estradiol (E2) and progesterone (P) levels were significantly lower in the POF group, but significant recovery was observed in terms of MVH and PCNA expression and E2 and P levels in the POF + PCs group. The apoptosis marker genes BAX and ROS were significantly increased in the POF group but were notably restored in the POF + PCs group. In addition, the expression of Sestrin2, an antiapoptotic protein, was significantly increased in the PCs treatment group, as were the upstream and downstream regulatory factors NRF2 and SOD2, and the indices of the Sestrin2 overexpression group were similar to those of the PCs treatment group. In summary, these findings suggest that PCs have potential as innovative therapeutic agents for preventing and treating POF by activating the protective SESTRIN2-NRF2 pathway against oxidative stress.

P-352 Investigating the association between DMPK CTG repeats and female hypogonadism in patients affected by Type 1 Myotonic Dystrophy

Abstract Study question Is DPKM CTG repeat size correlated with hypogonadism in female patients affected by type 1 Myotonic Dystrophy? Summary answer The size of CTG repeats in DPKM gene seems to be correlated with hypogonadism severity in female patients affected by type 1 Myotonic Dystrophy. What is known already Type 1 Myotonic Dystrophy (DMT1) is an autosomal dominant degenerative genetic disorder caused by CTG repeats in a single copy of DMPK gene. It is characterized by a broad phenotypic spectrum and a progressive course, involving primarily the muscular and endocrine systems. While DMT1 is a very well-known cause of male hypogonadism, the relationship between DMPK CTG repeats and female hypogonadism is almost unexplored yet. Study design, size, duration We performed a cross–sectional risk study in a cohort of 60 women, with a mean age of 31.5 years (18 to 45 years), referred to our Medical Genetics service for either suspected DMT1 or hypogonadism. The study duration was one year (from November 2022 to November 2023). Participants/materials, setting, methods We performed a DMPK–CTG analysis to investigate a possible correlation between CTG and hypogonadism. Based on the analysis, 11 women were diagnosed with DMT1 with associated hypogonadism, 11 with DMT1 without hypogonadism, 22 did not show DMPK expansion but were affected by hypogonadism, while the remaining 16 showed neither DMPK expansion nor hypogonadism. The patients affected by DMT1 performed standard karyotyping, FRAXA and FRAXE genetic testing to exclude alternative genetic causes of premature ovarian failure. Main results and the role of chance Our results, are indicative that the severity of hypogonadism in DMT1 patients may correlate with the size of CTG repeats. DMPK CTG repeats ranging from 150 – 1000 (range E2) were associated with milder forms of hypogonadism, while the patients having DMPK expansions within the E3 range (1000 CTG repeats or more) showed hypergonadotropic hypogonadism with both infertility and failure of medically assisted reproduction. Limitations, reasons for caution The female patients showing neither DMPK expansion nor hypogonadism did not perform other genetic analysis except DMPK-CTG testing. Moreover, due to the low sample size, our results did not reach statistical significance for a definitive conclusion. Wider implications of the findings To the best of our knowledge, this is the first study to show a possible correlation between female hypogonadism and DMPK CTG repeat size in female patients with type 1 myotonic dystrophy. These results, although preliminary, represent a first step in understanding a hitherto misrecognized clinical consequence of the disease. Trial registration number Not applicable

FRI283 Fragile X Gene Mutations Alter GnRH Neuron And Ovarian Innervation With Consequences On Reproductive Function

Abstract Disclosure: P.A. Villa: None. N. Lainez: None. C.R. Jonak: None. S. Berlin: None. I. Ethell: None. D. Coss: None. Mutations in the Fragile X messenger ribonucleoprotein 1 gene (FMR1) cause Fragile X syndrome, the most common cause of inherited mental disability, attributed to the loss of the Fragile X messenger ribonucleoprotein (FMRP). FMR1 mutations also comprise a majority of the genetic causes of premature ovarian failure (POF) in women, mechanisms of which are unknown. We utilized the Fmr1KO mouse model to determine the role of the FMR1 gene in the reproductive system in females. Similarly to what occurs in women harboring the mutation, our data demonstrate that Fmr1KO female mice experienced premature reproductive senescence, where reproductive function stopped at p163; in contrast, wild type controls stopped reproducing at p263. Since POF in women can occur due to either an insufficient pool of primordial follicles or early depletion of follicles, we quantified primordial follicles in 3 week old females and determined there was no difference between KO and controls. We then determined that Fmr1KO females had a higher number of corpora lutea at 8 weeks of age and gave birth to larger litters, suggesting that KO females recruit more follicles in each estrus cycle. Given that follicle stimulating hormone (FSH) and luteinizing hormone (LH) are essential in ovarian function, we analyzed gonadotropin levels in Fmr1KO females and determined they have higher serum LH and FSH. Inhibin b, progesterone, and testosterone levels were also higher, suggesting that high gonadotropins do not result from the lack of feedback. Ovariectomy determined that the hypothalamus drives high LH in KO females, while increased FSH is dependent on the ovaries. Increased vasculature in the corpora lutea and higher sympathetic innervation of developing follicles may increase steroid hormone production and drive increased serum FSH levels. To investigate the hypothalamic role in increased LH, we determined that a majority of gonadotropin-releasing hormone (GnRH) neurons express FMRP, which also have a higher number of GABAergic synapses in Fmr1KO females. Given that GABA is excitatory to GnRH neurons, alterations in afferent synaptic input can affect GnRH and LH secretion. LH pulsatile analysis determined higher LH pulse frequency in both unmodified and ovariectomized females, indicating an increase in GnRH neuron activity. In summary, our results reveal hypothalamic and ovarian contributions to the reproductive phenotype of Fragile X mutation carriers that may lead to early menopause. Presentation: Friday, June 16, 2023

Mitochondrial DNA Depletion syndrome 1 (MNGIE type) - a rare cause of premature ovarian failure?

Searchable abstracts of presentations at key conferences in endocrinology ISSN 1470-3947 (print) | ISSN 1479-6848 (online)

Menopausal symptoms and quality of life in female survivors treated with hematopoietic stem cell transplantation.

To assess the severity of menopausal symptoms and the correlation among different quality of life questionnaires and compare the quality of life of patients who underwent hematopoietic stem cell transplantation (HSCT) for hematological disorders with the norm group in order to facilitate personalized and directed therapeutic intervention for patients. We recruited women who had premature ovarian failure (POF) after HSCT for hematologic diseases in the gynecological endocrinology outpatient clinic of Peking University People's Hospital. Women with HSCT were included in the study if they had 6 months of spontaneous amenorrhea with serum follicle-stimulating hormone levels greater than 40 mIU/mL taken 4 weeks apart. The patients who had other causes of POF were excluded. During the survey, all women were required to fill out the questionnaires [Quality of Life Questionnaire (MENQOL), Generalized Anxiety Disorder-7 (GAD-7), Patient Health Questionnaire-9 (PHQ-9), and 36-item Short-Form (SF-36)] online. We analyzed the severity of menopausal symptoms, anxiety, and depression in Participants. In addition, differences on the SF-36 scale scores between the study group and norm groups were examined. In total, 227 (93.41%) patients completed the survey and were analyzed. The severity of all symptoms is "none and mild" in MRS, MENQOL, GAD-7, and PHQ-9. On the MRS, the most common symptoms were irritability, physical and mental exhaustion, and sleep problems. The severest symptoms were sexual problems (53, 73.82%), followed by sleep problems (44, 19.38%) and mental and physical exhaustion (39, 17.18%). In the MENQOL, the most common symptoms were psychosocial and physical symptoms. The severest symptoms were sexual symptoms (35, 48.75%) followed by psychosocial symptoms (23, 10.13%). Moderate-severe scores were shown in 11.89% (27) and 18.72% (42) cases in the GAD-7 and PHQ-9, respectively. Based on SF-36, in comparison with the norm group, the HSCT participants had higher vitality scores and lower role physical, physical functioning, and role emotional scores aged 18-45. In addition, the HSCT participants had lower mental health scores aged 18-25, and lower general health scores aged 25-45. No strong correlation was observed between questionnaires in our study. Overall, menopausal symptoms are milder in female patients after HSCT. There is no single scale that comprehensively assesses the patient's quality of life after HSCT. We need to assess the severity of various symptoms in patients using different scales.

Umbilical Cord-Derived Mesenchymal Stem Cells for the Treatment of Infertility Due to Premature Ovarian Failure.

Females belonging to the reproductive age group may face challenges regarding infertility or miscarriage due to conditions such as premature ovarian failure (POF). It is the conditionthat happens when a female's ovaries stop working before she is 40. The majority of the causes of POF cases are idiopathic. Other reasons includegenetic disorders (Turner's syndrome, bone morphogenetic protein 15 (BMP15) mutation, galactosemia, mutation of forkhead box protein L2 (FOXL2), growth differentiation factor-9 (GDF9), mutation of luteinizing hormone (LH) and follicle-stimulating hormonereceptors (FSHR), etc.), enzymatic mutation such as aromatase, autoimmune disorders (Addison's disease, vitiligo, systemic lupus erythematosus, myasthenia gravis, autoimmune thyroiditis, autoimmune polyglandular syndrome, etc.), vaccination, and environmental factors (cigarette smoking, toxins, and infections). Manyattempts have been made to treat POF by various methods. Some of the methods of treatment include hormone replacement therapy (HRT), melatonin therapy, dehydroepiandrosterone (DHEA) therapy, and stem cell therapy. Stem cell therapy has proven to be the most efficient form for treating POF as compared to all other options. Umbilical cord-derived mesenchymal stem cells (UC-MSCs) are the best among the other sources of mesenchymal stem cells (MSCs) for the treatment of POF as they have a painless extraction procedure. They have a tremendous capacity for self-repair and regeneration, which helps them in restoring degenerated ovaries. This review includes informationon the causes of POF, its efficacious therapeutic approaches, and the impact of transplantation of human umbilical cord mesenchymal stem cells (hUCMSCs) as an option for the therapy of POF. Numerous studies conducted on stem cell therapy provethat it is an effective approach for the treatment of sterility.

Research Progress on the Relationship Between Mitochondrial Dysfunction and Premature Ovarian Failure from the Perspective of TCM Treatment

Premature ovarian failure (POF) is a prevalent yet challenging condition in gynecology. Its incidence rate has been steadily rising in recent years and is trending among younger age groups. The premature failure of ovarian function, which poses a serious threat to women’s physical and mental health, has emerged as a topic of concern in current research. At present, it is believed that the causes of premature ovarian failure are multifaceted, and its pathogenesis is complex, some of which are still not well-understood. Therefore, this article discusses the research progress of the pathogenesis of premature ovarian failure caused by mitochondrial dysfunction and serves as a reference for the prevention and further research of this condition from the perspective of traditional Chinese medicine treatment.

Progress in Traditional Chinese Medicine Syndrome Differentiation and Treatment of Premature Ovarian Failure

Premature ovarian failure has directly or indirectly caused unspeakable physical and mental suffering to many women because of its increasing incidence. People of all ethnic groups in the world are actively seeking the best method, but the results are only basically satisfactory. As far as treatment is concerned, Chinese and Western medical treatments have their own advantages and disadvantages. However, because of the variety of syndrome types in traditional Chinese medicine, there are many descriptions of a single disease syndrome, and there are few systematic generalizations and classifications. Therefore, the experience of traditional chinese medicine in the treatment of premature ovarian failure in recent 10 years is classified and summarized. Traditional chinese medicine believes that the root cause of premature ovarian failure is the disorder of viscera, and the pathological sign is phlegm dampness and blood stasis. Therefore, the treatment should be based on kidney, syndrome differentiation and treatment of multiple viscera, taking into account phlegm dampness and blood stasis, combined with periodic therapy and acupuncture treatment, striving to nourish the kidney, regulate the heart, liver, spleen and lungs, and smooth phlegm dampness and blood stasis, so as to achieve the effect of prevention and treatment.

P–541 Identification of novel variants and candidate genes in women with familial idiopathic premature ovarian failure using whole-exome sequencing

Abstract Study question Is it possible to identify a genetic cause of familial premature ovarian failure (POF) with whole-exome sequencing (WES)? Summary answer Whole-exome sequencing is the most efficient strategy to identify probably pathogenic mutations in different genes in pathologies of polygenic etiology such as premature ovarian failure. What is known already Premature ovarian failure is the loss of ovarian function before the age of 40, and it is a common cause of infertility in women. This pathology has a heterogeneous etiology. Some chromosomal and genetic alterations have been described, and could explain approximately 20% of cases. However, in most patients the origin remains unknown. Recent studies with next-generation sequencing (NGS) have identified new variants in candidate genes related with premature ovarian insufficiency (POI) or premature ovarian failure (POF). These genes are not only involved in processes such as folliculogenesis, but also with DNA damage repair, homologous recombination, and meiosis. Study design, size, duration Fourteen women, from 7 families, affected by idiopathic POF were included in the study from October 2019 to September 2020. Seven POF patients were recruited when they came to our clinic to undergo assisted reproductive treatment. In the anamnesis, it was found that they had relatives with a diagnosis of POF, who were also recruited for the study. The inclusion criteria were amenorrhea before 38 years old and analytical and ultrasound signs of ovarian failure. Participants/materials, setting, methods WES was performed using TrusightOne (Illumina®). Sequenced data were aligned through BWA tool and GATK algorithm was used for SNVs/InDel identification. VCF files were annotated using Variant Interpreter software. Only the variants shared by each family were extracted for analysis and these criteria were followed: (1) Exonic/splicing variants in genes related with POF or involved in biological ovarian functions (2) Variants with minor allele frequency (MAF) ≤0.05 and (3) having potentially moderate/strong functional effects. Main results and the role of chance Seventy-nine variants possibly related with the POF phenotype were identified in the seven families. All these variants had a minor allele frequency (MAF) ≤0.05 in the gnomAD database and 1000 genomes project. Among these candidate variants, two were nonsense, six splice region, one frameshift, two inframe deletion and 68 missense. Thirty-two of the missense variants were predicted to have deleterious effects by minimum two of the four in silico algorithms used (SIFT, PolyPhen–2, MutationTaster and PROVEAN). All variants were heterozygous, and all the families carried three or more candidate variants. Altogether, 43 probably damaging genetic variants were identified in 39 genes expressed in the ovary and related with POF/POI or linked to ovarian physiology. We have described genes that have never been associated to POF pathology, however they may be involved in key biological processes for ovarian function. Moreover, some of these genes were found in two families, for example DDX11, VWF, PIWIL3 and HSD3B1. DDX11 may function at the interface of replication-coupled DNA repair and sister chromatid cohesion. VWF gene is suggested to be associated with follicular atresia in previous studies. PIWIL3 functions in development and maintenance of germline stem cells, and HSD3B1 is implicated in ovarian steroidogenesis. Limitations, reasons for caution Whole-exome sequencing has some limitations: does not cover noncoding regions of the genome, it also cannot detect large rearrangements, copy-number variants (large deletions/duplications), mosaic mutations, mutations in repetitive or high GC rich regions and mutations in genes with corresponding pseudogenes or other highly homologous sequences. Wider implications of the findings: WES has previously shown to be an efficient tool to identify genes as cause of POF, and has demonstrated the polygenic etiology. Although some studies have focused on it, and many genes are identified, this study proposes new candidate genes and variants, having potentially moderate/strong functional effects, associated with POF. Trial registration number Not applicable

TRDMT1 participates in the DNA damage repair of granulosa cells in premature ovarian failure.

The molecular mechanisms underlying premature ovarian failure, which seriously impacts the physical and psychological health of patients, are not fully understood. Here, we present the role of TRDMT1 in reactive oxygen species-induced granulosa cells death, which is considered an important cause of premature ovarian failure. We found that reactive oxygen species were increased in a H2O2 dose-dependent manner and accompanied by the nuclear shuttling of TRDMT1, increased DNA damage and increased apoptosis of granulosa cells. In addition, reactive oxygen species-induced granulosa cells apoptosis could be prevented by the antioxidant N-acetylcysteine or overexpression of TRDMT1. Furthermore, DNA repair following reactive oxygen species induction was severely impaired/enhanced in TRDMT1 mutants, which exhibited reduced/increased RNA m5C methylation activity. Altogether, our results reveal a novel role of TRDMT1 in the regulation of premature ovarian failure through the repair of reactive oxygen species-triggered DNA damage in granulosa cells and provide an improved understanding of the mechanisms underlying granulosa cells apoptosis, which could potentially be useful for future clinical treatments of premature ovarian failure.

Overactive Reproductive Axis Due to Fragile X Gene Mutation

Women carrying a pre-mutation or mutation of the Fragile X mental retardation gene (FMR1) comprise the largest portion of premature ovarian failure (POF) cases due to known genetic factors. FMR1 mutation causes Fragile X syndrome, the most common cause of inherited mental impairment. The mutation inhibits the expression of the fragile X mental retardation protein (FMRP), a ubiquitously expressed mRNA binding protein. The specific molecular mechanism(s) leading to premature ovarian failure in Fragile X carriers are not known. Here, we utilize the complete KO mouse model, to mimic the lack of FMRP in Fragile X mutations and analyze the hypothalamic-pituitary-gonadal axis to uncover causes of POF due to FMR1 mutation. Consistent with mutations in human population, KO females experience early cessation of reproductive function and stop having litters at 150 days of age, compared to controls that stop reproducing at 250 days of age. Since POF can be caused by either insufficient pool of primordial follicles or by increased recruitment in each cycle and early depletion, we analyzed ovaries at 3 weeks of age and determined that the FMR1 KO mice had the same number of primordial follicles when compared to the controls, suggesting that POF is not due to a deficit in primordial follicles. However, at 8 weeks of age, FMR1 KO ovaries had higher number of corpora lutea, and KO females had larger litters, indicating that FMR1 KO mice have more follicles recruited in each estrous cycle. FMR1 KO mice have higher FSH, which corresponds to the high FSH in women. Serum estradiol levels and inhibin b expression levels were unaffected by FMR1 mutation suggesting normally functioning negative feedback signals from the ovaries. Analyses of hypothalamic gene expression demonstrated elevated GnRH mRNA in KO mice. To further investigate alterations is hypothalamic protein levels, western blot analyses determined that FMR1 KO mice have higher levels of NMDAR1 and higher levels of GABAA receptor G2 subunit. Dual label immunofluorescence analyses revealed higher number of NMDAR1 and GABAA receptors specifically in GnRH neurons of FMR1 KO mice when compared to control, suggesting that GnRH neurons themselves are affected by FMR1 mutation. Given that both glutamate and GABA can activate GnRH neurons, alterations in the number of these receptors can potentially cause hyperactivity in the HPG axis at the hypothalamic level leading to elevated FSH and the subsequent POF. In summary, our results reveal a potential mechanism of premature ovarian failure in Fragile X mutation carriers.

Update on Female Fertility Preservation

Cancer is the second leading cause of death for women under 40. Survival rates are increasing due to earlier diagnosis and advanced treatment. One of the side effects of treatment is premature ovarian failure. This is why fertility preservation is part of the initial care of patients, allowing for a better quality of life after the disease. The main causes of premature ovarian failure are iatrogenic (mainly chemotherapy and radiotherapy) in more than a third of cases, idiopathic, genetic and autoimmune. This preservation of fertility should be offered as soon as possible, ideally before gonadotoxic treatments. The same is true when the cause is non-iatrogenic because the age of the woman has a determining role on the quality of the gametes obtained and therefore on the results. The different techniques are embryo vitrification, oocyte vitrification and freezing of ovarian tissue, to be combined when possible with ovarian blockage and/or ovarian transposition. The most common technique is oocyte vitrification, which allows oocytes to be conserved, although this is not possible in pre-pubescent girls and in cases of urgent treatment. In these cases the freezing of the ovarian tissue is the solution.

SIAH1 Promotes Granulosa Cells Senescence and Contributes to Premature Ovarian Failure by Reducing the Stability of TRF2

Background: Reactive oxygen species (ROS) exposure is an important trigger for granulosa cell (GC) senescence, which is considered to be an important cause of premature ovarian failure (POF). However, the cellular mechanisms that mediate this process needs further exploration. Methods: We respectively detected the ROS level and senescence level in the ovarian tissues and granulosa cells in POF rats. We also treated granular cells with H2O2 and antioxidant N-acetylcysteine (NAC) to detect the level of ROS and downstream protein levels, as well as the level of cell senescence. Findings: In the present, we observed that ROS levels were increased in the ovarian tissues of POF models in vivo, follicular GCs of POF patients and cyclophosphamide (CTX) pretreated GCs. In addition, ROS induced by H2O2 treatment was shown to significantly increase in SIAH1 expression, TRF2 reduction and senescence in GCs, which could be diminished by treatment with the Antioxidant N-acetylcysteine (NAC) or SIAH1 downregulation. Moreover, TRF2 was co-localized with SIAH1 in the cytoplasm, and SIAH1 was able to facilitate the ubiquitination and degradation of TRF2 via the ubiquitin-proteasome pathway, further leading to abnormalities in the telomeres of GCs. Interpretation: Our results indicate that ROS induces telomere abnormalities in GCs by promoting SIAH1-mediated TRF2 degradation, which causes cellular senescence, and finally contributes to POF. Funding Statement: This research was supported by the Natural Science Foundation of China (grant no. 81672913/81871343), the Social Development Project of Jiangsu (grant no. BE2018693/BE2017698), Natural Science Foundation of Jiangsu Province (grant no. BK20181226), the Jiangsu Provincial Medical Youth Talent (QNRC2016460, FRC201788) Declaration of Interests: The authors declare no conflict of interest. Ethics Approval Statement: The study was approved by the Reproductive Medical Center of the Fourth Affiliated Hospital of Jiangsu University, and the research protocol was approved by Jiangsu University Ethics Committee.

An adolescent girl with premature ovarian failure, Graves\u2019 disease, and chronic urticaria: a case report

BackgroundPremature ovarian failure is characterized by amenorrhea, hypoestrogenism, and hypergonadotropinism, and occurs in women under 40 years of age. The prevalence of premature ovarian failure in women younger than 20 years of age is only 0.01%. Immune disorders are one of the causes of premature ovarian failure. Graves’ disease and chronic urticaria are also associated with immune disorders.Case presentationWe report a case of a 15-year-old Han Chinese girl with premature ovarian failure complicated by Graves’ disease and chronic urticaria. She experienced menarche at 13 years of age and presented with amenorrhea after 7 months of irregular menstruation. Laboratory examinations indicated hypoestrogenism and hypergonadotropinism. Ultrasound imaging revealed that her uterus and ovaries were small in size. Gene and antibody tests related to premature ovarian failure returned negative results. Both thyroid peroxidase autoantibody and thyrotropin receptor antibody were positive.After reviewing the literature on the relationship between these three diseases and immune disorders, our patient was diagnosed as having atypical autoimmune polyglandular syndrome. After taking small doses of estrogen for 6 months, the size of her uterus increased, and her psychological anxiety was relieved.ConclusionsWe report a case of an unusual association of premature ovarian failure, Graves’ disease, and chronic urticaria. This case presents an atypical combination of adolescent autoimmune polyglandular syndrome, which is worthy of the attention of clinicians and presents an important lesson for them. Our case highlights that premature ovarian failure in adolescents requires long-term follow-up and medical treatment as well as psychological counselling.

Phytoestrogen genistein hinders ovarian oxidative damage and apoptotic cell death-induced by ionizing radiation: co-operative role of ER-\u03b2, TGF-\u03b2, and FOXL-2

Radiotherapy is a well-known cause of premature ovarian failure (POF). Therefore, we investigated the molecular influence of genistein (GEN) on the ovarian reserve of rats exposed to ϒ-radiation. Female Sprague Dawley rats were exposed to a 3.2 Gy γ-radiation to induce POF and/or treated with either GEN (5 mg/kg, i.p.) or Ethinyl estradiol (E2; 0.1 mg/kg, s.c.), once daily for 10 days. GEN was able to conserve primordial follicles stock and population of growing follicles accompanied with reduction in atretic follicles. GEN restored the circulating estradiol and anti-Müllerian hormone levels which were diminished after irradiation. GEN has potent antioxidant activity against radiation-mediated oxidative stress through upregulating endogenous glutathione levels and glutathione peroxidase activity. Mechanistically, GEN inhibited the intrinsic pathway of apoptosis by repressing Bax expression and augmenting Bcl-2 expression resulted in reduced Bax/Bcl-2 ratio with subsequent reduction in cytochrome c and caspase 3 expression. These promising effects of GEN are associated with improving granulosa cells proliferation. On the molecular basis, GEN reversed ovarian apoptosis through up-regulation of ER-β and FOXL-2 with downregulation of TGF-β expression, therefore inhibiting transition of primordial follicles to more growing follicles. GEN may constitute a novel therapeutic modality for safeguarding ovarian function of females’ cancer survivors.

Genetic Explanations for Fertility Disorders

This article reviews the genetic testing of infertility disorders. Genetic abnormalities can lead to disturbances of sexual development and hamper reproduction by influencing gamete production and maturation, fertilization, and embryonic development. Until now, the vast majority of detectable genetic abnormalities causing infertility were chromosomal abnormalities in both males and females. However, the number of monogenetic disorders, which play a role in disturbing fertility, such as single gene defects or complex disorders involving multiple genes in combination with environmental factors, is increasing. The common genetic causes of infertility in males are sex chromosomal abnormalities, Robertsonian translocations, cystic fibrosis transmembrane conductance regulator (CFTR) mutations, and Y chromosome microdeletions. In females, Turner syndrome, genetic causes of premature ovarian failure, such as fra(X) and complex disorders like polycystic ovary syndrome (PCOS), and endometriosis are the most prevailing. However, insight into the total impact of genetic abnormalities is increasing. This is because a growing number of mutations are detected among subfertile patients with an unknown etiology. Genetics is one of the most important yet under-emphasized causes of subfertility. Improved understanding of the genetics of infertility holds promise to define the etiology and counsel cases that were previously diagnosed with idiopathic infertility.

Differential Expression of DUB Genes in Ovarian Cells Treated with Di-2-Ethylhexyl Phthalate.

Premature ovarian failure (POF) is defined as loss of ovarian function in women less than 40 years of age. The causes of POF are diverse and include environmental factors. Di-2-ethylhexyl phthalate (DEHP) is one factor that may cause POF. The ubiquitin-proteasome system maintains intracellular balance by promoting or inhibiting protein degradation. To investigate the differential expressions of deubiquitinating enzyme (DUB) genes in patients with POF, we developed two in vitro POF models by treating A2780 or OVCAR5 with DEHP. Using these models, a multiplex RT-PCR system for DUB genes was applied to identify biomarkers by comparing expression patterns and DUB mRNA levels; multiplex RT-PCR results were validated by qRT-PCR and Western blotting analyses. Observed differential expression levels of several DUB genes including USP12, COPS5, ATXN3L, USP49, and USP34 in A2780 and OVCAR5 cells at the mRNA and protein levels suggest that they should be investigated as potential biomarkers of POF.

Prematür Ovaryen Yetmezlikli Hastalarda Karyotip Değerlendirmesi

INTRODUCTION: Chromosome anomalies are one of the major causes of premature ovarian failure and the importance of chromosome analysis in reproductive management has been confirmed. Numerical and structural chromosome anomalies, especially structural anomalies of the X chromosome, X-autosome translocations and X-chromosome aneuploidies, are the chromosome anomalies most commonly described in the literature. In this study, we aimed to evaluate the frequency and type of chromosomal anomalies in the patients with premature ovarian failure admitted to our clinic and to discuss the findings in the light of current literature and to provide guidance to new studies. METHODS: The files of the patients, who were diagnosed with premature ovarian failure between 2002 and 2017, were screened from the archive of the division of reproductive endocrinology and infertility in the department of obstetrics and gynecology at our center. 65 patients were included in the study. Information about age, smoking, alcohol use, age at menarche and menapose, hormone replacement therapy usage, additional disease and obstetric history were obtained from files. The laboratory results of the patients were obtained from files. FSH, LH, estradiol, prolactin, TSH, fT3, fT4, Anti-TPO, Anti-TG, TRAB, cortisol, ANA, Insulin, fasting blood glucose, LDL, HDL, triglyceride, total cholesterol, anti-Mullerian hormone levels were recorded from files. The karyotype results were recorded from files. RESULTS: The mean age at diagnosis was 32.6 years.The mean body mass index was 23.4(kg/m²). 60(92.3%) had normal karyotype(46+XX). 5(7.7%) had abnormal karyotype(4 had 46+XX/45+X and 1 had 46+XY/45+X). The mean value of fT3 is significantly higher in cases with normal karyotype(p: 0.019). DISCUSSION AND CONCLUSION: Considering the high rate of X chromosome loss in patients with premature ovarian failure, we can say that premature ovarian failure manifests itself in a wide spectrum. In conclusion, it can be said that cytogenetic studies should be evaluated routinely in cases with premature ovarian failure regardless of age.

Premature ovarian failure and tissue engineering.

Premature ovarian failure (POF) usually happens former to the age of 40 and affects the female physiological state premenopausal period. In this condition, ovaries stop working long before the expected menopausal time. Of diagnostic symptoms of the disease, one can mention amenorrhea and hypoestrogenism. The cause of POF in most cases is idiopathic; however, cancer therapy may also cause POF. Commonly utilized therapies such as hormone therapy, in-vitro activation, and regenerative medicine are the most well-known treatments for POF. Hence, these therapies may be associated with some complications. The aim of the present study is to discuss the beneficial effects of tissue engineering for fertility rehabilitation in patients with POF as a newly emerging therapy.

Hedgehog pathway inhibition causes primary follicle atresia and decreases female germline stem cell proliferation capacity or stemness

BackgroundFollicle depletion is one of the causes of premature ovarian failure (POF) and primary ovarian insufficiency (POI). Hence, maintenance of a certain number of female germline stem cells (FGSCs) is optimal to produce oocytes and replenish the primordial follicle pool. The mechanism that regulates proliferation or stemness of FGSCs could contribute to restoring ovarian function, but it remains uncharacterized in postnatal mammalian ovaries. This study aims to investigate the mechanism by which inhibiting the activity of the hedgehog (Hh) signaling pathway regulates follicle development and FGSC proliferation.Methods and resultsTo understand the role of the Hh pathway in ovarian aging, we measured Hh signaling activity at different reproductive ages and the correlation between them in physiological and pathological mice. Furthermore, we evaluated the follicle number and development and the changes in FGSC proliferation or stemness after blocking the Hh pathway in vitro and in vivo. In addition, we aimed to explain one of the mechanisms for the FGSC phenotype changes induced by treatment with the Hh pathway-specific inhibitor GANT61 via oxidative stress and apoptosis. The results show that the activity of Hh signaling is decreased in the ovaries in physiological aging and POF models, which is consistent with the trend of expression levels of the germline stem cell markers Mvh and Oct4. In vitro, blocking the Hh pathway causes follicular developmental disorders and depletes ovarian germ cells and FGSCs after treating ovaries with GANT61. The proliferation or stemness of cultured primary FGSCs is reduced when Hh activity is blocked. Our results show that the antioxidative enzyme level and the ratio of Bcl-2/Bax decrease, the expression level of caspase 3 increases, the mitochondrial membrane potential is abnormal, and ROS accumulate in this system.ConclusionsWe observed that the inhibition of the Hh signaling pathway with GANT61 could reduce primordial follicle number and decrease FGSC reproductive capacity or stemness through oxidative damage and apoptosis.