Potential Reproductive Toxicity Research Articles

Benzyl butyl phthalate promotes ferroptosis in Sertoli cells via disrupting ceruloplasmin-mediated iron balance.

Widespread environmental contamination with benzyl butyl phthalate (BBP) has raised concerns due to its high potential for bioaccumulation and male reproductive toxicity. However, the mechanisms underlying BBP-induced male reproductive damage remain unclear. As the adjacent Sertoli cell-formed blood-testis barrier (BTB) creates a privileged niche for spermatogenesis and may serve as the first target of reproductive toxicants, we mainly focused on the detrimental effect of BBP on Sertoli cells and the BTB in this study. C57BL/6 mice were administered BBP via oral gavage at doses ranging from 0 to 400mg/kg/day for 60 consecutive days. A comprehensive investigation was performed to estimate testicular BBP levels, sperm parameters, histological alterations, functional permeability of the Sertoli cell-based BTB, and ferroptosis in mice. Isolated Sertoli cells were further used to explore and validate the role of ferroptosis in BBP-induced BTB disruption. The results showed that permeation of BBP into the testis induced reduction in sperm quantity and quality, accompanied by fractured BTB ultrastructure, compromised permeable 'fence' functions of BTB, decreased expressions of tight junction proteins (TJP1 and OCLN) and paracellular transepithelial electrical resistance (TER) of Sertoli cells. Moreover, BBP exposure significantly increased intracellular iron content, promoted lipid peroxidation and activated ferroptosis in the testis of mice and primary Sertoli cells, which was involved in BBP-induced disruption of BTB integrity and function as confirmed by the ferroptosis inhibitors. In mechanism, BBP specifically downregulated the intracellular iron exporter ceruloplasmin (CP) level to inhibit Fe2+ export and the oxidization of Fe2+ into less toxic Fe3+, thus exacerbating ferroptosis in Sertoli cells. Overexpression of CP significantly suppressed ferroptosis and alleviated BBP-induced BTB disruption. These findings reveal the role of CP-mediated iron homeostasis in regulating Sertoli cell ferroptosis and BTB function, providing new insights into the mechanisms of BBP-related reproductive toxicity.

Black Garlic Extract Modulates Endothelin Expression and Ovulatory Function in Monosodium Glutamate Treated Rats.

Monosodium glutamate (MSG), a widely used food additive, has been associated with various health concerns, including potential reproductive toxicity. This study investigated the protective effects of black garlic (BG) ethanol extract against MSG-induced ovarian damage in rats. Thirty-two female rats in estrus were randomly divided into four groups (n = 8 per group): control (saline), BG (250 mg/kg BW), MSG (4 mg/g BW), and BG+MSG (combined treatment). Treatments were administered daily for 14 days. Ovarian tissues were collected for histopathological, immunohistochemical (IHC), and biochemical analyses. Histopathological examination revealed a significant reduction in cystic follicles in the BG+MSG group compared to the MSG group (p < 0.0001). IHC analysis showed decreased immunoreactivity of endothelin-1 and endothelin-2 in the BG+MSG group compared to the MSG group (both p < 0.01). Biochemical assays demonstrated significantly increased follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol levels in the BG+MSG group compared to the MSG group (all p < 0.05), while progesterone levels were significantly lower in the MSG group compared to the BG+MSG group (p < 0.05). These findings suggest that BG ethanol extract may mitigate MSG-induced ovarian dysfunction in rats by alleviating degenerative changes in follicles and modulating hormonal levels. This study provides insights into potential natural interventions for MSG-related reproductive toxicity.

Effect of different doses of cyclobenzaprine-HCl on the reproductive health of female Wistar rats

Background Women often experience significant burdens both at home and in their professional lives, which can lead to physical and mental strain. To manage these challenges, medications such as cyclobenzaprine-HCl, originally developed as a tricyclic antidepressant but reintroduced as a muscle relaxant, are sometimes used. Its effectiveness in treating muscle spasms, localized pain, and limited range of motion in acute musculoskeletal conditions has been well documented. Objective The objective of this study was to explore the potential consequences of cyclobenzaprine-HCl on reproductive health by assessing its influence on female Wistar rats. Materials and methods The study was conducted on 18 adult female Wistar rats, divided into three groups: a control group receiving distilled water and two treated groups with two doses − a low-dose group receiving 1.5 mg/kg of cyclobenzaprine-HCl and a high-dose group receiving 3 mg/kg, administered orally for 30 days. We assessed the impact of cyclobenzaprine-HCl on serum hormonal levels (progesterone, estradiol, luteinizing hormone, follicle-stimulating hormone), oxidative stress markers (glutathione, superoxide dismutase, catalase, malondialdehyde), histopathological changes in ovaries and uteri, and DNA stability using the comet assay. Results and conclusion The study revealed that cyclobenzaprine-HCl caused hormonal imbalances distinguished by significant increase in estradiol and follicle-stimulating hormone levels, and significant decreases in progesterone and luteinizing hormone levels. Dissruptions in oxidative stress markers were characterized by elevated malondialdehyde and reduced glutathione levels. Histopathological abnormalities included degeneration, deformed follicles, congested blood vessels, and necrosis in the ovarian tissue. In addition, diffused eosinophil infiltration, pyknotic nuclei, glandular hyperplasia, necrosis, and hypercellularity in the uterine tissue. DNA instability was observed in both dosage groups as evidenced by fragmented DNA in the shape of comets. These findings underscore the potential reproductive toxicity of cyclobenzaprine-HCl in female rats, suggesting a need for caution in its use considering its possible adverse effects on reproductive health.

Urinary profiles of bisphenol S derivatives and their exposure pathway analysis in maternal and infant populations of Beijing

Bisphenol S (BPS) derivatives have potential reproductive developmental toxicity and have been found in the environment and in breast milk. The level of infant exposure and the source are currently unknown. In this study, we investigated BPS and six derivatives (together referred to as BPs) in urine samples from mothers and infants, indoor dust, breast milk and infant formula in Beijing, China. BPS, diphenyl sulfone (DPS) and 4-allyloxy-4′-hydroxydiphenyl sulfone (BPS-MAE) were the main BPs. Notably, the concentration of DPS in infants’ urine was higher than that of BPS, which warrants attention. Infants have higher daily intake levels than mothers. Exclusively breastfed infants have a higher risk of BPs exposure than exclusively formula-fed infants. For exclusive breastfed infants, the contribution of individual BPs through breast milk was 23.2% to 93.6%. While for exclusively formula-fed infants, the contribution of individual BPs through infant formula was 30.5% to 70.3%. The contribution of individual BPs through indoor dust was no more than 10%. The results suggesting that infants can be exposed to BPs through other pathways. This is the first comprehensive assessment of maternal and infants exposure to BPS derivatives, providing insights into the sources of infant exposure.

Development of hybrid models by the integration of the read-across hypothesis with the QSAR framework for the assessment of developmental and reproductive toxicity (DART) tested according to OECD TG 414

The governing laws mandate animal testing guidelines (TG) to assess the developmental and reproductive toxicity (DART) potential of new and current chemical compounds for the categorization, hazard identification, and labeling. In silico modeling has evolved as a promising, economical, and animal-friendly technique for assessing a chemical's potential for DART testing. The complexity of the endpoint has presented a problem for Quantitative Structure-Activity Relationship (QSAR) model developers as various facets of the chemical have to be appropriately analyzed to predict the DART. For the next-generation risk assessment (NGRA) studies, researchers and governing bodies are exploring various new approach methodologies (NAMs) integrated to address complex endpoints like repeated dose toxicity and DART. We have developed four hybrid computational models for DART studies of rodents and rabbits for their adult and fetal life stages separately. The hybrid models were created by integrating QSAR features with similarities-derived features (obtained from read-across hypotheses). This analysis has identified that this integrated method gives a better statistical quality compared to the traditional QSAR models, and the predictivity and transferability of the model are also enhanced in this new approach.

Association between exposure to per- and polyfluoroalkyl substances and missed miscarriage: A hospital-based case-control study in Shanghai, China

Per- and polyfluoroalkyl substances (PFAS) are persistent environmental pollutants linked to reproductive disruptions. This study investigated the relationship between PFAS exposure and missed miscarriage in a hospital-based, case-control study in Shanghai, China. There were 393 women in our research, including 198 cases and 195 controls. Concentrations of 30 PFAS in plasma were quantified using HPLC-MS/MS, and 15 PFAS were detected at a rate greater than 90 percent. PFOA, L-PFOS, and PFOS isomers were significantly higher in the cases than those in the controls. 13 PFAS showed significant positive associations with miscarriage risk after adjustment for confounders, particularly PFOA (OR: 2.99, 95 % CI: 1.96–4.68) and various PFOS isomers. BKMR analysis confirmed higher overall PFAS levels were associated with increased miscarriage risk. These findings highlight the potential reproductive toxicity of PFAS and underscore the need for further investigations and regulatory actions to mitigate PFAS exposure in pregnant women.

Associations between per- and polyfluoroalkyl substances and menstrual cycle regularity in reproductive-aged female: A cross-sectional study

Menstrual cycle characteristics constitute one of the significant female fertility indicators. Previous epidemiological studies have shown that exposure to environmental chemicals could affect menstrual cycle characteristics, but the knowledge remains limited overall. Per- and polyfluoroalkyl substances (PFAS) have been identified as potential reproductive toxicants, while previous studies mainly focused on several legacy PFAS chemicals but generally failed to explore the outcomes from exposure to a complex mixture of both legacy and emerging PFAS. Besides, the modification effect of physical activity is rarely considered. In the present study, we explored the associations of exposure to a suite of legacy and emerging PFAS and menstrual cycle regularity as well as the potential modification by physical activity based on a pre-conception cohort in Shanghai (China) with the participation of 1001 reproductive-aged women. A total of 20 PFAS chemicals with detection frequency >80 %, which were derived from the PFAS exposure profile of the same population in our previous study, were included in the confounder-adjusted logistic regression and Bayesian kernel machine regression (BKMR) analysis. In individual PFAS analysis, after adjustment of the covariates, ∑2m-PFOS (the sum of all perfluoro-dimethylhexane sulfonates) was significantly associated with menstrual cycle irregularity with an odds ratio (OR) of 1.35 (95 % confidence interval, CI: 1.09, 1.67) as well as long cycles (OR = 1.37; 95 % CI: 1.08, 1.70). In addition, a significant positive association was also found between perfluoro-n-nonanoic acid (PFNA) and long cycles (OR = 1.40; 95 % CI: 1.06, 1.86). No significant associations were found between the PFAS mixture and the menstrual cycle characteristics as revealed by BKMR analysis, while the significant association between ∑2m-PFOS and menstrual cycle irregularity was also observed in the mixture exposure model. Subgroup analysis stratified by physical activity level showed that the associations between ∑2m-PFOS and menstrual cycle irregularity as well as long cycles were more pronounced in the inactive physical activity subgroup. This study suggested that branched PFOS (i.e., ∑2m-PFOS) might act as the predominant risk factor for menstrual cycle irregularity, and physical activity could influence the risks.

Copper Oxide Nanoparticles Impair Mouse Preimplantation Embryonic Development through Disruption of Mitophagy-Mediated Metabolism.

Copper oxide nanoparticles (CuONPs) have been widely applied, posing potential risks to human health. Although the toxicity of CuONPs on the liver and spleen has been reported, their effects on reproductive health remain unexplored. In this study, we investigate the effects of CuONPs on embryonic development and their potential mechanisms. Our results demonstrate that CuONPs exposure impairs mouse preimplantation embryonic development, particularly affecting the morula-to-blastocyst transition. Additionally, CuONPs were found to reduce the pluripotency of the inner cell mass (ICM) and mouse embryonic stem cells (mESCs). Mechanistically, CuONPs block autophagic flux and impair mitophagy, leading to the accumulation of damaged mitochondria. This mitochondrial dysfunction leads to reduced tricarboxylic acid (TCA) cycle activity and decreased α-ketoglutarate (α-KG) production. Insufficient α-KG induces the failure of DNA demethylation, reducing corresponding chromatin accessibility and consequently inhibiting ICM-specific genes expressions. Similar reduced development and inhibitions of pluripotency gene expression were observed in CuONPs-treated human blastocysts. Moreover, in women undergoing assisted reproductive technology (ART), a negative correlation was found between urinary Cu ion concentrations and clinical outcomes. Collectively, our study elucidates the mitophagy-mediated metabolic mechanisms of CuONPs embryotoxicity, improving our understanding of the potential reproductive toxicity associated with it.

Health risk assessment of toxic metals and DNA damage in somatic and germ cells by soil and groundwater of a major cement factory in Nigeria.

The waste generated from cement manufacturing is an important source of heavy metal contamination of groundwater and soil. This study investigated the concentration of toxic metals in the soil of a major cement factory and nearby groundwater. Ecological and carcinogenic risks of the metals were calculated. Potential reproductive toxicity and genotoxic effects of the samples were assessed in sex and somatic cells of male mice using sperm abnormalities and bone marrow micronucleus (MN) assays, respectively. Also, the serum ALP, ALT, AST, Total Testosterone (TT), Luteinizing Hormone (LH), and Follicle Stimulating Hormone (FSH); and liver SOD and CAT activities were measured in the treated mice. Cr, Cu, Ni, Zn, Mn, Cd, and Pb levels in the soil and groundwater exceeded the allowable maximum standard. Ingestion and dermal contact were the most probable routes of human exposure with children having about three times higher probability of exposure to the metals than the adults. Ni, Pb, and Cr presented carcinogenic risks in children and adults. In the MN result, nuclear abnormalities in the studied mice especially micronucleated polychromatic erythrocytes increased significantly (p < 0.05). Compared to the negative control, the ratio of PCE/NCE showed the cytotoxicity of the two samples. Data further showed a significant increase in the serum ALP, AST, and ALT while the liver CAT and SOD activities concomitantly decreased in the exposed mice. Sperm morphology result showed that the samples contained constituents capable of inducing reproductive toxicity in exposed organisms, with alterations to the concentrations of TT, LH, and FSH. Toxic metal constituents of the samples were believed to induce these reported reproductive toxicity and genotoxic effect. These results showed the environmental pollution caused by cement factory and the potential effects the pollutants might have on exposed eukaryotic organisms.

Bisphenol AP inhibits mouse oocyte maturation in vitro by disrupting cytoskeleton architecture and cell cycle processes

Bisphenol A (BPA) is among the extensively researched environmental endocrine-disrupting chemicals (EDCs), and its utilization is restricted owing to the detrimental impacts it has on human health. Bisphenol AP (BPAP) is one of the alternatives to BPA, but the influence of BPAP on human health has not been elucidated. The objective of the current research was to determine the influence of BPAP exposure on the in vitro maturation of mouse oocytes and to explore its potential reproductive toxicity. BPAP exposure was found to inhibit polar body extrusion during mouse oocyte maturation, resulting in an arrest at the metaphase I stage of meiosis. Exposure to BPAP led to sustained activation of BubR1, preventing the degradation of both Securin and Cyclin B1. Mechanistically, BPAP exposure disrupts spindle assembly and chromosome alignment. Levels of acetylated α-tubulin were significantly elevated in BPAP-treated oocytes, reflecting decreased spindle stability. Exposure to BPAP also induced DNA damage and impaired DNA damage repair. In addition, BPAP exposure altered histone modification levels. In summary, this investigation suggests that exposure to BPAP can influence cytoskeletal assembly, interfere with cell cycle progression, induce DNA damage, alter histone modifications, and ultimately impede oocyte meiotic maturation. This investigation enhances understanding of the impact of bisphenol analogs on female gametes, underscoring that BPAP cannot be considered a reliable replacement for BPA.

Risk substance identification of asphalt VOCs integrating machine learning and network pharmacology

Asphalt releases volatile organic compounds (VOCs) during paving processes, posing risks to workers and the environment. The complex composition of asphalt and the evolving of VOCs present challenges in accurately assessing their potential environmental and health impacts using traditional experimental approaches. This study aimed to develop a robust computational framework integrating machine learning and network pharmacology to predict the risks from the asphalt VOCs. The results show that the MACCS+XGBoost model achieved the highest predictive performance, with an accuracy of 0.85, balanced accuracy of 0.84, sensitivity of 0.83, specificity of 0.84, and F1-score of 0.84 in the external validation. The network pharmacology analysis revealed that the identified VOCs with reproductive toxicity potential may disrupt key processes such as spermatogenesis, ovarian function, and hormonal regulation, providing mechanistic insights into their potential impacts. This advancement supports a proactive approach to environmental protection and fosters the transition towards a more sustainable, low-carbon transportation.

Bisphenol H exposure disrupts Leydig cell function in adult rats via oxidative stress-mediated m6A modifications: Implications for reproductive toxicity

Bisphenol H (BPH) has emerged as a potential alternative to bisphenol A (BPA), which has been curtailed for use due to concerns over its reproductive and endocrine toxicity. This study investigates whether BPH exerts antiandrogenic effects by impairing Leydig cell function, a critical component in testosterone production. We administered orally BPH to adult male rats at doses of 0, 1, 10, and 100 mg/kg/day for 7 days. Notably, BPH treatment resulted in a dose-dependent reduction in testicular testosterone levels, with significant decreases observed at ≥ 1 mg/kg/day. Additionally, BPH affected the expression of key genes involved in steroidogenesis and cholesterol metabolism, including Nr5a1, Nr3c4, Lhcgr, Scarb1, and Star, at higher doses (10 and/or 100 mg/kg/day). The study also revealed alterations in antioxidant gene expression (Sod2 and Cat) and modulation of m6A-related genes (Ythdf1–3 and Foxo3) and their proteins. Through MeRIP-qPCR analysis, we identified increased m6A modifications in Scarb1 and Star genes following BPH exposure. In vitro experiments with primary Leydig cells confirmed that BPH enhanced oxidative stress and diminished testosterone production, which were partially mitigated by antioxidant vitamin E supplementation and Ythdf3 knockdown. Meanwhile, simultaneous administration of BPH and vitamin E to primary Leydig cells partially counteracted BPH-induced alterations in the Ythdf3 expression. Our findings underscore a novel mechanism by which BPH disrupts Leydig cell function through the oxidative stress-m6A modification-autophagy pathway, raising concerns about its potential reproductive toxicity.

Potential endocrine-disrupting effects of iprodione via estrogen and androgen receptors: evaluation using in vitro assay and an in silico model

This study was conducted to provide evidence, using in vitro and in silico testing methods, regarding the adverse effects of iprodione, a representative dichlorophenyl dicarboxamide fungicide, on the endocrine system. In the present study, we used the HeLa9903 stably transfected transactivation assay (OECD TG 455), 22Rv1/MMTV_GR‒KO androgen receptor transcriptional activation assay (OECD TG 458), and toxicity prediction using VEGA QSAR. Our results showed that iprodione had no estrogen receptor antagonistic or androgen receptor agonistic effects; however, iprodione was determined to be an estrogen receptor agonist (log PC10 value is less than − 9) and androgen receptor antagonist (log IC30 value is − 4.58) without intrinsic toxicity against the human cell lines used in this study. VEGA QSAR was used to evaluate five substances with structures similar to that of iprodione. Among them, four chemicals were found to have positive androgen receptor and aromatase activities and have been observed to be developmental toxicants. These results suggest that iprodione regulates steroid hormone receptor interactions and is a potential reproductive toxicant.

Poweromin X Ten, a polyherbal formulation improves male sexual function: In vivo and network pharmacology study.

Poweromin X Ten (PXT) is a polyherbal formulation, traditionally used to enhance male sexual function. However, the safety and benefits of PXT have not been scientifically evaluated. Therefore, the present study investigated the toxicity and aphrodisiac potential of PXT in male rats and explored its principal mechanisms of action. Male Wistar rats were orally administered PXT (50 or 100 mg/kg) for 28 days, and sexual activity parameters, including latency and frequency of mounting and intromissions, were studied. The reproductive toxicity and spermatogenic potential were also examined. Furthermore, dopamine and serotonin levels in brain regions associated with sexual activity were assessed. Network analysis was used to identify the key bioactive compounds and their core targets involved in their beneficial actions. Treatment with PXT improved sexual activity in male rats, as evidenced by reduced mounting and intromission latency and a significant increase in mount frequency. Moreover, PXT exhibited spermatogenic potential and did not induce reproductive toxicity. Notably, treatment with 50 mg/kg PXT elevated dopamine levels in median preoptic area and hypothalamus. Pathway analysis indicated that PXT primarily modulated the PI3K-Akt, calcium, and MAPK signalling pathways to enhance male sexual function. Network analysis identified macelignan, β-estradiol, testosterone, and paniculatine as key bioactive components of PXT, which likely act through core targets, such as androgen receptor (AR), Mitogen-activated protein kinase 3 (MAPK3), epidermal growth factor receptor (EGFR), estrogen receptor 1 (ESR1), and vascular endothelial growth factor (VEGF) to facilitate the improvement of male sexual function. Study results suggest that PXT is a safer alternative with aphrodisiac and spermatogenic potential. These effects are partly attributed to the enhanced dopamine levels in the brain. Furthermore, this study provides insights into the specific signalling pathways and bioactive compounds that underlie the improvements in male sexual function associated with PXT.

Extended one-generation reproductive toxicity study of food-grade titanium dioxide E171 with emphasis on reproductive and endocrine endpoints

Food-grade titanium dioxide E171 was administered in feed to Sprague Dawley rats in an extended one-generation reproductive toxicity (EOGRT) study (OECD Test 443). The dosed diet (0, 100, 300, or 1000 mg/kg body weight/day) started 10 weeks before mating and continued throughout the study. After weaning, pups were allocated to Cohorts 1 A/1B (to assess reproductive toxicity), 2 A/2B (to assess developmental neurotoxicity), and 3 (to assess developmental immunotoxicity); in addition, Cohort 1B was mated to produce an F2 generation and satellite F0 animals were evaluated for colonic aberrant crypt foci (ACF). In F0 animals, there were no systemic toxicity or reproductive effects, no treatment-related histopathological changes, and no ACF in the colon. Serum estradiol or testosterone concentrations were not changed in F0 or F1 animals. No pre-/postnatal developmental changes related to treatment were noted in F1 animals, and the reproductive performance of F1 Cohort 1B animals was unaffected. F2 pups showed no abnormalities in pre- or postnatal development (postnatal days 4–8). No treatment-related developmental neurotoxicity was observed in Cohorts 2 A/2B. Although no treatment-related immunotoxicity was observed in Cohort 3, the positive control did not induce the expected response; this segment of the study will be repeated. Analyses of blood and urine showed negligible systemic absorption of E171 from the gastrointestinal tract upon dietary ingestion. The no observed adverse effect level (NOAEL) for parental systemic toxicity, reproductive toxicity, offspring toxicity, and developmental neurotoxicity was considered 1000 mg/kg body weight/day. For developmental immunotoxicity, a NOAEL was not determined owing to insufficient T-cell-dependent antibody response in the positive control. Our study provides robust data on the reproductive toxicity and preneoplastic potential of E171.

Raman-guided exploration of placental microplastic exposure: Unraveling the polymeric tapestry and assessing developmental implications

The prevalence of microplastics in human tissues and their potential reproductive toxicity have been increasingly documented, yet their appearance in the placenta and the impact of microplastic exposure on human fertility and pregnancy remains uncertain. Utilizing an inVia™ confocal Raman microspectroscopy by Renishaw equipped with a detection threshold as low as 0.25 µm, our study examined the microplastics in the placentas of 50 women post-delivery and investigated their correlations with gestational age, and neonatal length and weight. We found that 40 microplastic particles were identified across 31 of 50 placentas, averaging 2.35 ± 1.25 µm in size and ranging from 1.03 to 6.84 µm. Seven distinct polymer types were detected, with PTFE, PS, and ABS being the most prevalent. Notably, no significant difference across the normal, PTFE, and PS groups for all demographic variables examined was identified, nor as pathological alterations of placental tissues. In conclusion, our findings demonstrate the presence of seven microplastic polymers in human placentas, with PTFE, PS, and ABS being the most prevalent. However, maternal and neonatal parameters were not affected, and further studies are necessary to elucidate the effects of microplastics on developmental outcomes and fetal health.

GRP78/IRE1 and cGAS/STING pathway crosstalk through CHOP facilitates iodoacetic acid-mediated testosterone decline

Iodoacetic acid (IAA) is an emerging unregulated iodinated disinfection byproduct with high toxicity and widespread exposure. IAA has potential reproductive toxicity and could damage male reproduction. However, the underlying mechanisms and toxicological targets of IAA on male reproductive impairment are still unclear, and thus Sprague-Dawley rats and Leydig cells were used in this work to decode these pending concerns. Results showed that after IAA exposure, the histomorphology and ultrastructure of rat testes were abnormally changed, numbers of Leydig cells were reduced, the hypothalamic-pituitary-testis (HPT) axis was disordered, and testosterone biosynthesis was inhibited. Proteomics analyses displayed that oxidative stress, endoplasmic reticulum stress, and steroid hormone biosynthesis were involved in IAA-caused reproductive injury. Antioxidant enzymes were depleted, while levels of ROS, MDA, 8-OHdG, and γ-H2A.X were increased by IAA. IAA triggered oxidative stress and DNA damage, and then activated the GRP78/IRE1/XBP1s and cGAS/STING/NF-κB pathways in Leydig cells. The two signaling pathways constructed an interactive network by synergistically regulating the downstream transcription factor CHOP, which in turn directly bound to and negatively modulated steroidogenic StAR, finally refraining testosterone biosynthesis in Leydig cells. Collectively, IAA as a reproductive toxicant has anti-androgenic effects, and the GRP78/IRE1 and cGAS/STING pathway crosstalk through CHOP facilitates IAA-mediated testosterone decline.

An in vitro testicular organoid model for the study of testis morphogenesis, somatic cell maturation, endocrine function, and toxicological assessment of endocrine disruptors

Male reproductive capacity has fallen considerably in recent decades; in addition, the incidence of testicular cancer has increased in many developed countries. The cause of this phenomenon is unknown, but environmental toxicants are considered a major contributing factor. To study potential reproductive toxicants, robust in vitro testis models are needed. We have recently established a porcine testis organoid system with a high resemblance to the architectures of innate testis tissue. Here, we further investigated the testis morphogenesis, cell maturation, and endocrine function of the testis organoids. We also challenged this system with abiraterone, a steroidogenic inhibitor, to validate its suitability as an in vitro platform for endocrine toxicology tests. Our results showed that the testis cells in the organoids reorganize into testis cordal structures, and the cordal relative areas increase in the organoids over time of culture. Moreover, the diameters and cell numbers per cross-section of the cordal structures increased over time. Interestingly, Sertoli cells in the organoids gradually underwent maturational changes by showing increased expression of androgen receptors, decreased expression of the anti-müllerian hormone, and formation of the blood-testis barrier. Next, we confirmed that the organoids respond to hormonal stimulation and release multiple sex hormones, including testosterone, estradiol, and progesterone. Finally, we showed that the production of testosterone and estradiol in this system can be inhibited in response to the steroidogenic inhibitor. Taken together, our organoid system provides a promising in vitro platform for male reproductive toxicology studies on testis morphogenesis, somatic cell maturation, and endocrine production.

Features, Potential Invasion Pathways, and Reproductive Health Risks of Microplastics Detected in Human Uterus.

Microplastics (MPs) are ubiquitous in global ecosystems and may pose a potential risk to human health. However, critical information on MP exposure and risk to female reproductive health is still lacking. In this study, we characterized MPs in human endometrium and investigated their size-dependent entry mode as well as potential reproductive toxicity. Endometrial tissues of 22 female patients were examined, revealing that human endometrium was contaminated with MPs, mainly polyamide (PA), polyurethane (PU), polyethylene terephthalate (PET), polypropylene (PP), polystyrene (PS), and polyethylene (PE), ranging from 2-200 μm in size. Experiments conducted in mice demonstrated that the invasion of the uterus by MPs was modulated either through diet-blood circulation (micrometer-sized particles) or via the vagina-uterine lacuna mode (larger particles reaching a size of 100 μm. Intravenous exposure to MPs resulted in reduced fertility and abnormal sex ratio in mouse offspring (P < 0.05). After 3.5 months of intragastric exposure, there was a significant inflammatory response in the endometrium (P < 0.05), confirmed by embryo transfer as a uterine factor leading to decreased fertility. Furthermore, human endometrial organoids cultured with MPs in vitro exhibited significantly apoptotic responses and disrupted growth patterns (P < 0.01). These findings raise significant concerns regarding MP contamination in the human uterus and its potential effects on reproductive health.

Poweromin X Ten, a polyherbal formulation improves male sexual function: In vivo and network pharmacology study

Introduction Poweromin X Ten (PXT) is a polyherbal formulation, traditionally used to enhance male sexual function. However, the safety and benefits of PXT have not been scientifically evaluated. Therefore, the present study investigated the toxicity and aphrodisiac potential of PXT in male rats and explored its principal mechanisms of action. Methods Male Wistar rats were orally administered PXT (50 or 100 mg/kg) for 28 days, and sexual activity parameters, including latency and frequency of mounting and intromissions, were studied. The reproductive toxicity and spermatogenic potential were also examined. Furthermore, dopamine and serotonin levels in brain regions associated with sexual activity were assessed. Network analysis was used to identify the key bioactive compounds and their core targets involved in their beneficial actions. Results Treatment with PXT improved sexual activity in male rats, as evidenced by reduced mounting and intromission latency and a significant increase in mount frequency. Moreover, PXT exhibited spermatogenic potential and did not induce reproductive toxicity. Notably, treatment with 50 mg/kg PXT elevated dopamine levels in median preoptic area and hypothalamus. Pathway analysis indicated that PXT primarily modulated the PI3K-Akt, calcium, and MAPK signalling pathways to enhance male sexual function. Network analysis identified macelignan, β-estradiol, testosterone, and paniculatine as key bioactive components of PXT, which likely act through core targets, such as androgen receptor (AR), Mitogen-activated protein kinase 3 (MAPK3), epidermal growth factor receptor (EGFR), estrogen receptor 1 (ESR1), and vascular endothelial growth factor (VEGF) to facilitate the improvement of male sexual function. Conclusion Study results suggest that PXT is a safer alternative with aphrodisiac and spermatogenic potential. These effects are partly attributed to the enhanced dopamine levels in the brain. Furthermore, this study provides insights into the specific signalling pathways and bioactive compounds that underlie the improvements in male sexual function associated with PXT.