Metabolic syndrome (MetS) contributes to male infertility, yet the impact of MetS-targeted therapies on fertility remains unclear. This study investigates the impact of obeticholic acid (OCA), a farnesoid X receptor (FXR) agonist, in a high-fat-diet (HFD)-induced MetS mouse model. MetS was induced in male mice by 12-week of HFD feeding. Mice then received OCA (30 mg/kg) or vehicle for 5 weeks. Sperm parameters, testicular and epididymal immune profiles, intestinal bile acid, gut microbiota, and epididymal gene expression were assessed. OCA treatment was associated with higher sperm progressive motility (PR) and lower proportions of epididymal IFN-γ-producing CD4+ T helper 1 (Th1) cells. OCA shifted bile acid profile and gut microbiota. Expression of H2bc24 in epididymis increased to levels comparable with control group. These findings indicate that OCA is associated with improved male reproductive parameters in MetS through modulation of bile acid signaling, immune response, and microbiota. OCA may represent a potential therapeutic approach for MetS- associated male infertility.
Oxidative stress is a primary driver of male infertility, underscoring the critical importance of effective antioxidant defense systems in maintaining reproductive function. Astaxanthin, a xanthophyll carotenoid with potent free radical–scavenging capacity and membrane-stabilizing properties, has garnered interest as a therapeutic agent for male reproductive health. Experimental evidence indicates that astaxanthin modulates cellular redox homeostasis, preserves mitochondrial integrity, and attenuates lipid peroxidation in spermatozoa and testicular cells. Through these mechanisms, it potentially maintains sperm structural integrity and functional competence under oxidative stress. Preclinical studies demonstrate that astaxanthin supplementation enhances sperm quality, supports mitochondrial bioenergetics, and protects against testicular oxidative injury. However, clinical evidence in men remains limited and heterogeneous, with variations in study populations, treatment durations, dosages, and outcome measures. Although some clinical investigations report improvements in semen parameters and markers of oxidative stress, others lack consistent findings. Consequently, these clinical data must be interpreted with caution. This narrative review synthesizes knowledge on the biological mechanisms through which astaxanthin influences male reproductive physiology, including oxidative regulation and stress response modulation. It evaluates existing data, identifies significant knowledge gaps, and highlights research priorities. To clarify the therapeutic utility of astaxanthin in male infertility, well-designed, large-scale randomized controlled trials featuring standardized formulations and robust clinical outcomes are required.
Hypertensive disorders of pregnancy (HDP) confer increased cardiovascular disease (CVD) risk postpartum. Screening methods targeting women with increased risk are lacking, limiting personalized follow-up. Circulating biomarkers may assist CVD risk stratification. We aimed to assess whether circulating biomarkers were associated with measures of cardiovascular structure and function following HDP and normotensive pregnancies (controls). We examined 275 women 1–3 years postpartum (176 following HDP and 99 parous controls). Circulating biomarkers (hs-CRP, non-HDL cholesterol, glucose, eGFR, cTnT, NT-proBNP, GDF-15, sHLA-G, SAA1) and cardiovascular function and structure measures were examined [pulse wave velocity (PWV), carotid intima-media thickness (CIMT) and echocardiography]. Group differences were tested by Mann-Whitney U test and chi-square. Associations were assessed by univariate regression analyses. A p-value < 0.05 was considered statistically significant. Women following HDP displayed significantly higher rates of hypertension (5
GPR30 (G-protein coupled receptor 30) plays a key role in regulating reproductive processes, particularly through estrogen signaling. Previous studies have linked GPR30 polymorphisms to female reproductive disorders such as infertility, recurrent pregnancy loss, endometriosis, leiomyoma, and breast or ovarian cancers. This study investigated, for the first time, the association between two GPR30 polymorphisms (rs3808350 and rs3808351) and recurrent implantation failure (RIF) in infertile women referred to the Royan Institute. A total of 300 Iranian women were divided into three groups: 100 with RIF, 100 fertile controls, and 100 women with successful ART outcomes as a second control group. The frequencies of rs3808350 and rs3808351 polymorphisms were analyzed using ARMS-PCR, and confirmed by Sanger sequencing. Statistical analysis was performed using Chi-square and Fisher’s exact tests (p ≤ 0.05). For rs3808350, genotype frequencies significantly differed across the three groups (p < 0.001), though allele frequencies did not (p = 0.107). Further comparison between the RIF and ART+ groups showed significant differences in both genotype (p = 0.005) and allele (p = 0.041) frequencies. For rs3808351, significant differences in both genotype and allele frequencies were observed among all groups (p < 0.001). These findings suggest that rs3808350 and rs3808351 polymorphisms in the GPR30 gene are significantly associated with increased risk of recurrent implantation failure.
To identify immune-related molecular targets in polycystic ovary syndrome (PCOS) and evaluate the role of integrin beta-2 (ITGB2) and the potential modulation by rutin. Follicular fluid cytokines from 30 women with PCOS and 30 control women were profiled. Transcriptomic (GSE34526) and DNA methylation (GSE138573) data were integrated to identify overlapping genes, followed by functional enrichment analyses, protein–protein interaction (PPI), and molecular docking analyses. LPS-stimulated granulosa cells, in the presence or absence of ITGB2 overexpression and rutin treatment, were assessed for gene expression, viability, cytokine secretion, and NF-κB activation. Women with PCOS had higher IL-5 but lower IL-1β and IL-12p70. ITGB2, ITGAM, and CD86 were upregulated and hypomethylated. Among these, ITGB2 bound rutin with the lowest energy (− 7.8 kcal/mol). Rutin reduced ITGB2 expression, restored viability, suppressed cytokine release, and inhibited NF-κB activation, even under ITGB2 overexpression. ITGB2 is associated with inflammatory responses in granulosa cells and may contribute to PCOS-related immune dysregulation via NF-κB activation. Rutin attenuates these inflammatory responses, supporting a potential involvement of the ITGB2/NF-κB axis as a therapeutic target. Not applicable.
Rheumatoid arthritis and its glucocorticoid-based treatment may adversely affect male reproductive function by disrupting spermatogenesis, steroidogenesis, and testicular homeostasis. Insulin-like peptide 3 (INSL3), a Leydig cell-derived peptide hormone and marker of Leydig cell functional capacity, may contribute to the maintenance of testicular function through its steroidogenic and anti-inflammatory effects. In the present study, we investigated the effects of prednisolone and INSL3, administered alone or in combination, on testicular structure and function in rats with adjuvant-induced arthritis. Adult male rats were allocated to seven groups: control, prednisolone, INSL3, arthritis, arthritis + prednisolone, arthritis + INSL3, and arthritis + prednisolone + INSL3. Testicular alterations were evaluated using histological, morphometric, immunohistochemical, ultrastructural, testosterone, and sperm analyses. Arthritis was associated with reduced testicular weight, decreased seminiferous tubule diameter and epithelial thickness, impaired sperm parameters, lower serum and intratesticular testosterone levels, increased testicular tumor necrosis factor-α and interleukin-6 immunoreactivity, and reduced 3β-hydroxysteroid dehydrogenase and proliferating cell nuclear antigen immunoreactivity. In arthritic rats, prednisolone or INSL3 alone was associated with partial improvement in selected testicular parameters, whereas combined treatment was associated with improvement across several inflammatory, morphometric, steroidogenic, and sperm-related parameters. These findings suggest that INSL3 treatment is associated with partial attenuation of inflammation-related testicular injury and improvement in selected reproductive parameters under inflammatory and glucocorticoid stress. Further studies are required to clarify whether these effects are mediated by direct intratesticular mechanisms or indirect systemic pathways.
Bisphenol A (BPA) exposure is associated with gestational diabetes mellitus (GDM); however, the underlying molecular mechanisms remain elusive. This study aimed to identify key genes mediating BPA exposure-related GDM through integrative bioinformatics analysis. We integrated BPA-related targets from multiple databases with GDM-associated genes derived from GeneCards and the GSE103552 dataset. Functional enrichment analysis was performed on the intersecting genes. A protein-protein interaction (PPI) network was constructed, and hub genes were identified using four topological algorithms. Machine learning approaches were employed for feature selection. Immune infiltration was assessed, and core gene expression was validated via reverse transcription quantitative polymerase chain reaction (RT-qPCR), additional GEO datasets, and single-cell RNA sequencing data. Thirty-four overlapping genes were identified and found to be enriched in the insulin signaling pathway, the AMPK pathway, and inflammatory responses. Following further screening, five core genes—HSPA8, NR3C1, ITGB1, NOTCH2, and POU5F1—were selected, demonstrating robust diagnostic performance. Immune analysis revealed significant correlations between hub genes and the infiltration levels of immune cells. Consistent expression patterns of ITGB1 and NOTCH2 were validated across additional GEO datasets. RT-qPCR confirmed the downregulation of HSPA8 and the upregulation of NR3C1 and NOTCH2 in GDM. Single-cell analysis corroborated cell-type-specific expression within the placenta. This study elucidates potential molecular links between BPA exposure and GDM, highlighting immune-metabolic dysregulation. The identified core genes may serve as biomarkers or therapeutic targets, thereby advancing precision environmental health strategies for pregnancy.
Polycystic ovary syndrome (PCOS), in which hyperandrogenism is a major feature, is the most prevalent endocrine disorder among women of reproductive age. Serum free testosterone (FT) reflects the biologically active fraction of circulating testosterone. Accurate assessment of FT allows for precise identification of hyperandrogenism and provides critical guidance for the diagnosis of PCOS. Women with PCOS and normal controls (NCs) were included in this study. Fasting serum samples were collected during the early follicular phase to evaluate key clinical parameters, and FT levels were quantified using liquid chromatography‒tandem mass spectrometry (LC‒MS/MS). The diagnostic value of FT for identifying PCOS was assessed using Spearman’s correlation analysis and receiver operating characteristic curves. A total of 202 women were included in the PCOS group, 121 were included in the NC group. Total testosterone (TT), free androgen index (FAI), and FT levels were significantly greater in the PCOS group than in the NC group. Both FAI and FT were positively correlated with body mass index (BMI), triglycerides, low-density lipoprotein cholesterol, fasting insulin and the homeostasis model assessment for insulin resistance; and were negatively correlated with high-density lipoprotein cholesterol. Following age and BMI adjustments, the area under the curve (AUC) for FT (AUC = 0.886) was greater than that for TT (AUC = 0.856) or FAI (AUC = 0.864). Compared with conventional assessments of TT and FAI, LC–MS/MS–based measurements of FT demonstrate superior diagnostic performance for identifying PCOS. Both FT and FAI are associated with obesity and the features of metabolic syndrome in PCOS patients.
Non-alcoholic fatty liver disease (NAFLD), a common metabolic disorder, is increasingly linked to impaired male reproductive health. This study investigates the therapeutic potential of adipose-derived mesenchymal stem cells (AD-MSCs) and their exosomes (AD-MSCs-Exo) in ameliorating NAFLD-induced testicular dysfunction in a murine model. Male C57BL/6 mice were divided into four groups: [n = 8 per group: control, high-fat diet (HFD), HFD + AD-MSCs, and HFD + AD-MSCs-Exo]. Sperm parameters according to the WHO Laboratory Manual, hormonal profiles via ELISA, and expression of genes involved in autophagy and spermatogenesis were assessed using qRT-PCR. HFD-induced NAFLD significantly reduced sperm count (P = 0.036), motility (P < 0.0001), viability (P = 0.023), and normal morphology (P = 0.015) while increasing DNA fragmentation (P = 0.004). Moreover, in the HFD group, LH and total testosterone levels decreased significantly (P = 0.0002 and P < 0.0001), whereas estradiol levels increased significantly (P = 0.0001) compared with controls. Expression of STAR was significantly downregulated (P = 0.0001); conversely, the relative fold changes of BECN1 (P = 0.0003), MAP-LC3b-a (P = 0.0002), and SQSTM-1/p62 (P = 0.005) were significantly enhanced in the HFD group compared to the controls. Treatment with AD-MSCs or AD-MSCs-Exo improved sperm quantity and quality and balanced hormonal levels. AD-MSCs showed slightly greater modulation of key autophagy genes (BECN1, MAP-LC3b-a, and SQSTM1/p62), whereas exosomes were more effective in hormonal restoration (especially testosterone) and in enhancing genes related to steroidogenesis (especially INHBB). AD-MSCs and their exosomes improve sperm-related abnormalities, hormonal imbalance, and testicular autophagy dysregulation in the NAFLD model. These findings support their potential as therapeutic agents for NAFLD-induced male infertility, highlighting translational relevance while noting the need for further clinical validation.
Polycystic ovary syndrome (PCOS) is a complex reproductive and metabolic disorder characterized by hyperandrogenism, anovulation, insulin resistance, and chronic low-grade inflammation. The LIF/JAK/STAT3 pathway is very important for coordinating folliculogenesis, endometrial receptivity, and metabolic homeostasis. The LIF–LIFR–gp130–JAK1–STAT3 signaling pathway supports the survival of granulosa cells, follicle recruitment, ovulation, implantation, and mitochondrial function under physiological conditions. In polycystic ovary syndrome (PCOS), hyperandrogenism, hyperinsulinemia, and pro-inflammatory cytokines destabilize the LIFR/gp130 heterodimer, impede LIF and JAK1 signaling, and redirect STAT3 activation towards JAK2-mediated IL-6/leptin-dominated inflammatory pathways. This pathological rewiring promotes hyperandrogenic steroidogenesis by increasing the levels of LHCGR and CYP17A1 and decreasing the levels of FSHR and CYP19A1. It also causes follicular arrest and decreases the ability of the endometrium to accept an embryo by deregulating the LIF/STAT3–MUC1–IGF1 axis. Epigenetic alterations, such as hypomethylation of the first intron of JAK2 and hypermethylation of the SOCS3 promoter, in conjunction with the participation of regulatory non-coding RNAs (miR-181a/b, miR-199a-5p, lncRNA H19, circRNAs), facilitate the amplification of JAK2/STAT3 signaling and the suppression of inhibitory feedback mechanisms. Dysregulated mitochondrial STAT3 leads to impaired oxidative phosphorylation, modified glycolytic pathways, heightened oxidative stress, and worsened insulin resistance. The connections between the MAPK/ERK, PI3K/AKT, TLR4/NF-κB, Wnt/β-catenin, and Hippo/Notch/YAP pathways make this pathological signaling network even stronger. The LIF/JAK/STAT3 axis is a key regulator and a promising target for precision therapies tailored to specific phenotypes in PCOS. Therapeutic strategies, encompassing preclinical JAK inhibitors, metabolic modulators, and clinically validated pathway-directed agents, underscore its potential as a pivotal therapeutic nexus.
Some studies have reported that low serum progesterone (P) levels on the day of frozen-thawed embryo transfer (FET) are associated with lower pregnancy rates following artificial endometrial preparation (AEP) cycles. Other studies, however, have found no consistent association between serum P levels and pregnancy outcomes. This prospective cohort study included 235 patients who underwent FET following AEP cycles with estradiol valerate and vaginal micronized P. All patients received either blastocyst- or cleavage-stage embryos derived from their own oocytes, without preimplantation genetic testing for aneuploidies. Blood samples were collected on the day of FET to measure serum P concentrations. Serum P levels on the day of embryo transfer did not differ significantly between patients who achieved CP and those who did not (12.76 ± 4.1 ng/mL vs. 11.9 ± 5.6 ng/mL; p = 0.3318). The optimal serum P cutoff level associated with higher CP rates was 10.6 ng/mL, with a sensitivity of 72
Conventional semen analysis provides limited information on sperm kinematic behavior in couples with unexplained infertility undergoing intrauterine insemination (IUI). We used a laboratory based, microscope mounted smartphone-assisted Computerized Sperm Counting and Tracking Analysis System (CSCTAS) to characterize sperm kinematics in prepared samples and compare them by ultrasound-confirmed clinical pregnancy after IUI. In this retrospective single-center Turkish cohort, 184 first IUI cycles between January 2018 and December 2019 were analyzed. Prepared samples were assessed under standardized Makler chamber conditions. Manual–CSCTAS agreement for total motility was assessed by Bland–Altman analysis, and single-variable receiver operating characteristic analyses were supplementary. Clinical pregnancy occurred in 29 cycles (15.8
Actin-like protein 7A (ACTL7A) has been suggested to participate in sperm head formation and oocyte activation. This study investigates the expression and localization of ACTL7A in men with globozoospermia and explores its association with sperm chromatin integrity. Semen samples were collected from 10 infertile men diagnosed with 100
This study aimed to investigate whether chronic endometritis (CE) affects pregnancy outcomes in non-assisted reproductive technology (non-ART) treatments, including timed intercourse (TI) and intrauterine insemination (IUI). This prospective observational study included 50 patients who underwent conventional infertility treatment at Yamagata University Hospital between April 2020 and March 2022. Endometrial tissue samples were collected during hysteroscopic screening and assessed for CE using CD138 immunostaining. CE was diagnosed when ≥ 5 CD138-positive plasma cells were identified in at least one of 10 randomly selected high-power fields. Pregnancy outcomes were compared between CE-positive and CE-negative groups, and generalized estimating equation analyses were performed to account for repeated treatment cycles within the same patient. Among the 50 participants, 27 were CE-positive and 23 were CE-negative, yielding a CE prevalence of 54.0
Polycystic ovary syndrome (PCOS) is characterized by oxidative stress (OS). This study reveals a novel epigenetic mechanism linking lysine acetyltransferase 5 (KAT5) and thrombospondin-1 (TSP-1) in PCOS pathogenesis. A meta-analysis of 50 clinical studies demonstrated elevated systemic oxidative markers (total oxidant status, nitric oxide, malondialdehyde) and reduced paraoxonase-1 levels in PCOS patients, alongside compromised total antioxidant capacity in follicular fluid. Bioinformatic analysis identified TSP-1 as a key downregulated gene in PCOS, and the hTFtarget database predicted KAT5 as an upstream regulator of TSP-1. In vitro, H₂O₂-induced oxidative stress in KGN granulosa cells suppressed TSP-1, KAT5, and H2AK5ac levels, which were restored by N-acetylcysteine (NAC). Mechanistically, KAT5 was found to regulate TSP-1 expression via H2AK5 acetylation at its promoter. Knockdown of KAT5 exacerbated oxidative stress, apoptosis, and senescence, while TSP-1 overexpression counteracted these effects. In a letrozole-induced PCOS rat model, ovarian tissues exhibited decreased H2AK5 acetylation, reduced KAT5 and TSP-1 expression, and impaired redox homeostasis. Interventions with N-acetylcysteine or TSP-1 ameliorated ovarian pathological changes, hormonal imbalances, oxidative stress, apoptosis, and cellular senescence. Collectively, these findings establish the KAT5-TSP-1 axis as a critical regulator of ovarian redox homeostasis, unveiling a novel epigenetic mechanism and proposing a dual-target therapeutic strategy for PCOS.
The TSC complex, formed by the binding of TSC2 with TSC1 and TBC1D7, plays an important role in the onset of endometrial-related diseases through the TSC-mTORC1 axis. However, the relationship between these proteins and adenomyosis has not been determined thus far. Here, we aimed to investigate the expression of TSC proteins in adenomyosis and determine the correlation between TSC expression and clinicopathologic parameters in patients with adenomyosis. 21 patients (age range, 40–50 years) with histologically diagnosed adenomyosis who underwent hysterectomy for nonendometrial disease were enrolled in this study. Specimens of healthy endometria were obtained from 21 patients (age range, 38–53 years) with cervical carcinoma in situ who underwent laparoscopy. The participants were interviewed via a standard questionnaire consisting of items pertaining to sociodemographic characteristics and reproductive history. The severity of dysmenorrhea and menorrhagia was quantified by means of the visual analogue scale and the menstrual pictogram, and preoperative hemoglobin levels were determined. Samples of serum and endometrial tissue were collected, TSC and VEGF expression was determined via immunofluorescence, and VEGF expression in the serum was quantified via ELISA. We found that, in patients with adenomyosis, TSC expression was significantly decreased during the secretory phase. Endometrial TSC1 and TSC2 expression was correlated with menstrual volume. Additionally, high levels of VEGFD increased the likelihood of moderate-to-severe menorrhagia in adenomyosis patients. Our results suggest that TSC is associated with clinical symptoms such as menorrhagia. In addition, VEGFD may be a potential quantitative predictor of the severity of menorrhagia in patients with adenomyosis.
Polycystic Ovary Syndrome (PCOS) is a heterogeneous endocrine-metabolic disorder characterized by ovulatory dysfunction, hyperandrogenism, and insulin resistance, in which mitochondrial dysfunction has been increasingly implicated. Mitochondria regulate energy metabolism and oxidative stress, with mitophagy maintaining cellular balance. Dysregulated mitophagy relates to PCOS metabolic issues like obesity and inflammation. This study analyzed gene expression datasets to find autophagy-related genes in PCOS, identifying AMFR, FABP5, and HMOX1 as key genes. We built a diagnostic model and confirmed their elevated expression in a hyperandrogenism-induced PCOS cell model, revealing potential small-molecule drugs targeting these genes. Our integrative bioinformatics analysis and systematic molecular experiments suggest that FABP5 and HMOX1 are potential PCOS diagnostic targets, with high-affinity compounds as therapies,highlighting the pathological relevance of mitophagy-related genes in PCOS.
This study aims to evaluate the protective effects of betaine (BTN), an antioxidant agent, against ovarian and uterine toxicity caused by favipiravir (FVP), an antiviral agent, through multifaceted damage pathways. A total of 28 Wistar rats were divided into four experimental groups: Control, BTN (250 mg/kg), FVP (200 mg/kg), and FVP + BTN. At the end of the 10-day protocol, blood samples were collected along with ovarian and uterine tissue samples. Subsequently, the samples were subjected to biochemical analysis, qRT-PCR analysis, histological evaluation, and immunohistochemical studies. These methods assessed oxidative stress, inflammatory, apoptotic, and autophagic responses, hormonal changes, and overall tissue damage. FVP administration significantly increased oxidative stress (MDA, p < 0.001), inflammation (NF-κB, TNF-α, IL-1β, p < 0.001), apoptosis (Caspase-3, Bax, p < 0.001), and autophagy (Beclin-1, LC3A, p < 0.001); while decreasing antioxidant levels, Bcl-2, and steroidogenesis genes (CYP11A1, CYP17A1, CYP19A1, p < 0.001), and reducing serum AMH, E2, FSH, and LH values (p < 0.001). When BTN was administered concomitantly, most of these changes were reversed, antioxidant and hormonal balance were significantly restored, and the increases in oxidative, inflammatory, and apoptotic markers were reduced (p < 0.05–0.001). Histologically, betaine preserved tissue integrity by mitigating ovarian and uterine damage caused by FVP (p < 0.05). FVP caused multifaceted oxidative, inflammatory, apoptotic, and hormonal damage in ovarian and uterine tissues, while BTN significantly reduced these effects, preserving tissue integrity and function.
This matched comparative study investigates whether unoperated ovarian endometriomas (OMAs) influence the prevalence of tubal pathology as detected by hysterosalpingography (HSG) in infertile women. A total of 266 infertile women with unilateral or bilateral OMAs ≥ 2 cm and 539 matched controls without endometriomas were included between January 2021 and October 2024. Diagnosis of OMAs was confirmed by transvaginal ultrasonography and, if necessary, MRI, and all participants underwent standardized HSG assessment. Tubal pathologies were classified into nine distinct categories. The OMA group showed significantly higher rates of tubal abnormalities (31.6
High-fructose corn syrup (HFCS) consumption has been increasingly associated with reproductive dysfunction, yet the potential protective role of natural probiotic products such as kefir remains insufficiently explored. This study investigated the effects of kefir supplementation on HFCS-induced testicular damage in juvenile male Wistar rats. Twenty-four four-week-old rats (≈ 100 g) were maintained under controlled environmental conditions and randomly assigned to four groups: Control, Kefir, HFCS, and HFCS+Kefir. Kefir was administered by gastric gavage at 1 mL/100 g body weight daily, while HFCS-55 was provided ad libitum in drinking water (20
