Reproductive toxicology最新文献

{"title":"Embryotoxicity analysis of anti-arrhythmia drugs amiodarone, dronedarone, and their metabolites using 3D gastruloid models.","authors":"Courtney Kehaulani Kurashima, Yusuke Marikawa","doi":"10.1016/j.reprotox.2025.109070","DOIUrl":"10.1016/j.reprotox.2025.109070","url":null,"abstract":"<p><p>Amiodarone and dronedarone are anti-arrhythmic drugs that are structurally related but differ in iodine content. Although contraindicated during pregnancy due to suspected embryotoxicity based on animal studies, their mechanisms of action and relevance to human development remain unclear. Here, we used gastruloids - 3D aggregates of mouse or human pluripotent stem cells that recapitulate axial elongation morphogenesis of early embryos - to investigate their developmental effects. In mouse gastruloids, both drugs and their major metabolites impaired growth and elongation at 1.5 - 3.0 µM. They also altered expression of genes involved in somite segmentation and retinoic acid biosynthesis. Notably, dronedarone down-regulated additional genes, and only amiodarone's morphological effects were alleviated by retinoic acid supplementation, suggesting distinct mechanisms of action. In human gastruloids, dronedarone induced abnormal convoluted morphology and disrupted gene expression at concentrations as low as 0.05 µM, whereas amiodarone showed effects at 2.0 µM, indicating greater sensitivity of the human model to dronedarone. Transcriptomic analyses revealed both overlapping and distinct gene expression changes between the two drugs. These results demonstrate that gastruloid-based assays can detect adverse effects of amiodarone and dronedarone at clinically relevant concentrations, as therapeutic plasma levels are approximately 1.3 - 2.6 µM for amiodarone and 0.15 - 0.30 µM for dronedarone. The study also provided mechanistic and human-relevant insights not attainable through traditional animal testing. Our findings underscore the utility of stem cell-based models for assessing human developmental toxicity, and support their use in evaluating safer alternatives for anti-arrhythmic therapy during pregnancy.</p>","PeriodicalId":21137,"journal":{"name":"Reproductive toxicology","volume":" ","pages":"109070"},"PeriodicalIF":2.8,"publicationDate":"2025-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145177928","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exposure to low-dose polystyrene nanoplastics impairs the estrous cycle by decreasing ovarian levels of steroidogenic acute regulatory protein and serum progesterone levels in rats.","authors":"Lethícia Valencise, Jorge Willian Franco de Barros, Ana Flávia Quiarato Lozano, Luan Reis Calixto, Daniel G Cyr, Wilma De Grava Kempinas","doi":"10.1016/j.reprotox.2025.109069","DOIUrl":"10.1016/j.reprotox.2025.109069","url":null,"abstract":"<p><p>Plastic can be fragmented into smaller pieces referred to as microplastics (< 5 mm), or nanoplastics (< 1 µm). These particles have been reported to cross biological barriers and cause oxidative stress damage in several tissue types. Given that female reproductive tissues are considered a target for such particles, our study aimed to evaluate the effects of polystyrene nanoplastics (PS-NP, 500 nm) at a low concentration (0.015 mg/d), on reproductive parameters of adult female Wistar rats. Animals (n = 10/group) were treated by gavage for 25 days with PS-NP diluted in distilled water at a concentration of 0.015 mg/d. The Control group received only distilled water (vehicle). We assessed weight gain, estrous cyclicity, sexual behavior and fertility, morphology of ovaries and uteri, immunostaining for StAR in the ovaries, and serum levels of the steroid hormones: estradiol and progesterone. Data was evaluated by Student's t-test, Mann-Whitney test, or Fisher's Exact test. Results were considered significantly different when P ≤ 0.05. The PS-NP group showed estrous cycle dysregulation, uterine inflammatory infiltration, increased uterus and pituitary weight, and decreased thyroid weight in the experimental conditions utilized. These findings are potentially due to the decrease in StAR expression in luteal cells, and consequent reduction of progesterone serum levels. These results indicate that nanoplastics act as endocrine disruptors impairing female endocrine and reproductive function, in a rodent model, and raise concern about outcomes after exposure to nanoplastics in other females and in adult women's reproductive health.</p>","PeriodicalId":21137,"journal":{"name":"Reproductive toxicology","volume":" ","pages":"109069"},"PeriodicalIF":2.8,"publicationDate":"2025-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145150718","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Melatonin alleviates Di-2-ethylhexyl phthalate induced male reproductive toxicity through inhibition of endoplasmic reticulum stress: Behavioral, biochemical, and morphological evidences","authors":"Jiten Singh ,&nbsp;Ashok Jangra ,&nbsp;Sapana Kushwaha ,&nbsp;Itishree Dubey ,&nbsp;Dinesh Dhingra ,&nbsp;Dinesh Kumar","doi":"10.1016/j.reprotox.2025.109068","DOIUrl":"10.1016/j.reprotox.2025.109068","url":null,"abstract":"<div><div>Di(2-ethylhexyl) phthalate (DEHP) is one of the most commonly used plasticizers known for its effect on reproductive systems. The underlying molecular mechanism of DEHP-induced male reproductive toxicity is still less explored. Melatonin (Mel), a neurohormone possessing antioxidant properties, has demonstrated reproprotective effect in various studies. In this study, the underlying molecular mechanisms of DEHP induced reproductive toxicity and reproprotective effect of melatonin were investigated. Adult male Wistar rats were randomly divided into four experimental groups: control, DEHP-500 mg/kg, DEHP+Mel-3 mg/kg and DEHP+Mel-10 mg/kg. Animals were treated with DEHP (500 mg/kg; <em>p</em>.<em>o</em>.) for 28 days. Melatonin was administered at doses of 3 and 10 mg/kg for the last 14 days. DEHP exposure impaired the sexual motivational behavior as well as copulatory behavior in rats, which was ameliorated by melatonin treatment. In addition, Mel reversed the changes in the gonadosomatic index along with sperm quality and quantity. DEHP reduced the serum testosterone level and increased the level of testicular nitrite, and MDA while, decreased the level of reduced glutathione. However, Mel mitigated testicular oxidative stress and restored the serum testosterone level in DEHP-exposed rats. Additionally, histopathology and scanning electron microscopy revealed that the administration of Mel attenuates the cellular alteration as evidenced by Johnsen’s index scores. Western blotting analysis showed that protein expression of C/EBP homologous protein (CHOP), caspase-12, and glucose-regulated protein (GRP-78) were found upregulated in DEHP-exposed rat testes, indicating ER-stress mediated cell death. Mel reversed DEHP-induced protein expression level of CHOP, GRP-78 and caspase-12 in testicular tissue. In conclusion, the findings of the present study showed that Mel could be an effective intervention in the treatment of DEHP-induced reproductive toxicity. Moreover, ER-stress mediated apoptotic cells death played a vital role in DEHP-induced male reproductive toxicity.</div></div>","PeriodicalId":21137,"journal":{"name":"Reproductive toxicology","volume":"138 ","pages":"Article 109068"},"PeriodicalIF":2.8,"publicationDate":"2025-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145150682","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"NAD+ precursors mitigate the in vitro and in vivo reproductive defects: Limitations and possible solutions","authors":"Nazli Pinar Arslan ,&nbsp;Zuleyha Akpinar ,&nbsp;Havva Aybek ,&nbsp;Meryem Doymus ,&nbsp;Gulsum Asilkan-Kaldik ,&nbsp;Nevzat Esim ,&nbsp;Mesut Taskin","doi":"10.1016/j.reprotox.2025.109067","DOIUrl":"10.1016/j.reprotox.2025.109067","url":null,"abstract":"<div><div>In mammalian cells, nicotinamide adenine dinucleotide (NAD⁺) participates in the regulation of diverse cellular processes such as ATP production, oxidative stress resistance, DNA repair, metabolic homeostasis, and inflammation. Due to these properties, exogenously applied NAD⁺ precursors (nicotinic acid, nicotinamide, nicotinamide riboside, and nicotinamide mononucleotide) can protect organs and cells of mammalian against detrimental effects of various stress factors and diseases. For instance, NAD⁺ and its precursors have critical importance for the <em>in vivo</em> and <em>in vitro</em> fertilization success of mammals. This review summarizes that the natural aging process, diseases, and toxic compounds cause the detrimental effects in the reproductive parameters of the <em>in vivo</em> models, such as the meiotic defects and the reductions in cellular NAD⁺ level, mitochondrial functions, sperm and oocyte quality, blastocyst and embryo formation rate, implantation success, whereas the intragastric, intraperitoneal or oral administration of NAD⁺ precursors prevents or attenuates these detrimental effects. Similarly, the supplementation of NAD⁺ precursors can protect the oocytes and sperms against the cryopreservation process, aging and toxic compounds in the <em>in vitro</em> and also enhances blastocyst and embryo formation <em>in vitro</em>. This review study also revealed that the ability of NAD⁺ precursors-loaded drug delivery systems to prevent reproductive defects has not yet been investigated in literature. Therefore, we recommend the development of NAD⁺ precursor-loaded drug delivery systems targeting reproductive system organs and/or cell organelles (mitochondria, endoplasmic reticulum and nucleus). To achieve this, hormone receptors in testicular and ovarian cells can be targeted. Similarly, triphenylphosphonium (TPP⁺) can be used to specifically target mitochondria.</div></div>","PeriodicalId":21137,"journal":{"name":"Reproductive toxicology","volume":"138 ","pages":"Article 109067"},"PeriodicalIF":2.8,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145107891","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Silica nanoparticles inhibit the development of mouse preantral follicle in vitro 3D culture via a decrease in cell junctions and an increase in cell apoptosis","authors":"Haoxiang Wang ,&nbsp;Rongrong Ye ,&nbsp;Tuba Latif Virk ,&nbsp;Qi Liu ,&nbsp;Yuguo Yuan ,&nbsp;Fenglei Chen","doi":"10.1016/j.reprotox.2025.109066","DOIUrl":"10.1016/j.reprotox.2025.109066","url":null,"abstract":"<div><div>Silica nanoparticles (SNPs) are promising nanomaterial with desirable chemical and physical properties for their applications in agriculture, biomedicine and others. There are serious concerns regarding the biosafety of SNPs. SNPs can induce follicular atresia, leading to ovarian toxicity. However, studying only oocytes or ovarian granulosa cells seems insufficient to fully reveal the impact of SNPs on the development of follicles. An <em>in vitro</em> 3D culture system of the follicles was established to investigate mouse ovarian toxicity of SNPs. Follicular development was evaluated through observation of follicular morphology, measurement of follicular diameter, number of mature follicles and oocyte maturation. The secretion of 17-β estradiol (E₂) was measured by ELISA. Microfilament distribution was examined by phalloidin-iFluor, and mitochondria distribution was detected by Mitotracker Red. The results showed that SNPs disrupted ovarian granulosa cell arrangement, inhibited their growth, and hindered follicular antrum formation and oocyte maturation. SNPs significantly increased cell apoptosis in ovarian granulosa cells and elevated <em>Caspase-3</em> mRNA level. Additionally, SNPs significantly reduced E₂ secretion and <em>STAR</em>, <em>CYP11A1</em>, and <em>CYP19A1</em> mRNA levels. Furthermore, SNPs caused mitochondria to distribute unevenly, forming clusters or even diffusely within the oocytes. SNPs also shortened the length and reduced the number of microvilli in the follicles. Meanwhile, both CX37 protein and mRNA level were significantly decreased in the oocytes. In summary, SNPs induced follicular atresia, at least partly, via inhibition of oocyte maturation and damage of the gap functions between the oocytes and ovarian granulosa cells. This study confirmed the toxicity of SNPs on follicles using <em>in vitro</em> 3D culture of preantral follicles, providing a new perspective for the study of SNP-induced ovarian toxicity.</div></div>","PeriodicalId":21137,"journal":{"name":"Reproductive toxicology","volume":"138 ","pages":"Article 109066"},"PeriodicalIF":2.8,"publicationDate":"2025-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145102857","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Inulin attenuates reproductive toxicity caused by prenatal and lactation GenX exposure through modulating the hypothalamic–pituitary–testicular (HPT) axis via the LH/cAMP/StAR pathway in male mice","authors":"Ya-Qi Chen ,&nbsp;Yu-Kui Chen ,&nbsp;Jin-Jin Zhang ,&nbsp;Qin-Yao Zhang ,&nbsp;Yu Liu ,&nbsp;Ji-Hong Wang ,&nbsp;Qi Wang ,&nbsp;Xiao-Li Xie","doi":"10.1016/j.reprotox.2025.109065","DOIUrl":"10.1016/j.reprotox.2025.109065","url":null,"abstract":"<div><div>With persistent and bioaccumulation concerns, hexafluoropropylene oxide dimer acid (HFPO-DA or GenX) has been detected in surface water, plants, and biota. However, its male reproductive toxicity is poorly understood. In this study, pregnant 8-week-old C57BL/6 J female mice (n = 36) were administered Milli-Q water, GenX (2 mg/kg/day), GenX with inulin (5 g/kg/day) or inulin by gavage until weaning (PND28) to evaluate the effects on the reproductive function of the male offspring. Early-life GenX exposure resulted in decreased sperm quality, impaired testicular tissue structure, and reduced serum testosterone and luteinizing hormone (LH) levels in male mice (<em>p</em> &lt; 0.05), suggesting reproductive toxicity and disturbance of the hypothalamic<img>pituitary<img>testicular axis. Consistent with the enrichment of the cAMP pathway shown by transcriptomics analysis, decreased protein expression of StAR, CYP11A1, CYP17A1, 3β-HSD, 17β-HSD and PKA (<em>p</em> &lt; 0.05) and overexpression of NPY (<em>p</em> &lt; 0.05) were also detected in the GenX group. Inulin alleviated testicular structure injury resulting from GenX exposure and increased serum testosterone and LH levels as well as protein expression in the cAMP pathway. Taken together, our results suggest that prenatal and lactation GenX exposure reduces testosterone synthesis in male mice by modulating the LH-mediated cAMP pathway, leading to male reproductive disorders, whereas inulin intervention might attenuate reproductive toxicity.</div></div>","PeriodicalId":21137,"journal":{"name":"Reproductive toxicology","volume":"138 ","pages":"Article 109065"},"PeriodicalIF":2.8,"publicationDate":"2025-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145092490","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Different types of nanoplastics exert varying degrees of toxicity on human sperm at semen-related concentrations","authors":"Yujie Wang ,&nbsp;Yanfan Cui ,&nbsp;Yixuan Li ,&nbsp;Tao Luo ,&nbsp;Jiyan Li","doi":"10.1016/j.reprotox.2025.109064","DOIUrl":"10.1016/j.reprotox.2025.109064","url":null,"abstract":"<div><div>Previous research on the toxicity of nanoplastics (NPs) on male reproduction have centered on polystyrene (PS), a diverse range of NPs types has been identified in human semen and testes, warranting further investigation. In the present study, we employed pyrolysis gas chromatography-mass spectrometry (Py-GCMS) to quantify the presence of multiple NPs in human semen. Subsequently, we assessed the impact of various NPs at semen-related concentrations on human sperm functions and physiological parameters. Polypropylene (PP, 0.19–33.85 μg/mL), polyethylene (PE, 3.53–865.55 μg/mL), and polyethylene terephthalate (PET, 305.74 μg/mL) were detected in human semen. Exposure to PE at concentrations of 0.5 and 1 mg/mL significantly reduced sperm viability, motility, penetration ability, and mitochondrial membrane potential (MMP), while also inducing oxidative stress. At 1 mg/mL, PET decreased sperm motility, penetration ability and MMP, though it did not affect viability or reactive oxygen species (ROS) levels. In contrast, PP exhibited no adverse effects on sperm functions and physiological parameters at semen-related concentrations. Our findings demonstrate that different types of NPs exert varying degrees of toxicity on human sperm at semen-related concentrations, with PE exhibiting the highest toxicity and PP showing no observable toxicity. Moreover, exposure to mixture of NPs more accurately reflects human daily exposure scenarios. We evaluated the effects of mixed NPs on sperm function and observed that, with increasing co-incubation time, the NPs mixture induced more pronounced impairments in sperm viability and other parameters compared to exposure to any single type of NPs.</div></div>","PeriodicalId":21137,"journal":{"name":"Reproductive toxicology","volume":"138 ","pages":"Article 109064"},"PeriodicalIF":2.8,"publicationDate":"2025-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145092455","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Editorial - Special issue: Environmental chemicals and the mammary gland.","authors":"Laura N Vandenberg, Suzanne E Fenton, Sofie Christiansen","doi":"10.1016/j.reprotox.2025.109062","DOIUrl":"10.1016/j.reprotox.2025.109062","url":null,"abstract":"","PeriodicalId":21137,"journal":{"name":"Reproductive toxicology","volume":" ","pages":"109062"},"PeriodicalIF":2.8,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145086902","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Integrative computational dissection of monobenzyl phthalate spermatotoxicity: High-affinity binding disrupts ACE2-CYP17A1 coregulation in non-obstructive azoospermia","authors":"Guangqiang Zhu ,&nbsp;Chunlin Tan ,&nbsp;Yugen Li","doi":"10.1016/j.reprotox.2025.109063","DOIUrl":"10.1016/j.reprotox.2025.109063","url":null,"abstract":"<div><div>Phthalates, as typical environmental endocrine disruptors, can lead to reproductive toxicity by disrupting human endocrine homeostasis with their metabolites. However, the complexity of human co-exposure to multiple metabolites, confounding biases in traditional studies, and reverse causality issues obscure the causative contributions and pathogenic mechanisms of key toxic metabolites. This study employs a multi-omics integrative strategy, leveraging Mendelian Randomization (MR) to identify Monobenzyl phthalate (MBzP) as a causative risk factor for testicular damage (β = 1.26, P = 0.002). Focusing on Non-obstructive azoospermia (NOA), we integrated network toxicology to identify 15 shared molecular targets. By combining interpretable machine learning, we discovered that core targets ACE2/CYP17A1 are significantly overexpressed in Sertoli cells. Gene Set Enrichment Analysis (GSEA) of single genes revealed critical pathway differentiation: low expression states activate spermatogenesis pathways, while high expression drives inflammatory-apoptotic networks (TGF-β, p53 pathways, interferon response). Molecular docking and dynamics simulation (MDS) confirmed that MBzP forms stable complexes with ACE2 (binding energy = −7.2 kcal/mol) and CYP17A1 (binding energy = −7.5 kcal/mol), interfering with their physiological functions through allosteric effects. This study for the first time elucidates the MBzP-ACE2/CYP17A1 interaction-inflammatory/apoptotic cascade activation-spermatogenesis inhibition molecular axis, providing new targets for precise intervention in male infertility.</div></div>","PeriodicalId":21137,"journal":{"name":"Reproductive toxicology","volume":"138 ","pages":"Article 109063"},"PeriodicalIF":2.8,"publicationDate":"2025-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145081446","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effects of PD-1 inhibitors on ovarian function-based on two-sample mendelian randomization and visualization experimental validation","authors":"Jiaxin Zheng ,&nbsp;Na Zhu ,&nbsp;Chang Liu ,&nbsp;Xiang Li ,&nbsp;Keming Zhang ,&nbsp;Zhuo Yang ,&nbsp;Danbo Wang ,&nbsp;Bo Liu","doi":"10.1016/j.reprotox.2025.109057","DOIUrl":"10.1016/j.reprotox.2025.109057","url":null,"abstract":"<div><div>Although immune checkpoint inhibitors (ICIs) have transformed cancer treatment by improving survival, their ovarian safety remains uncertain. This study combined Mendelian randomization (MR) and experimental validation to assess the impact of PD-1 inhibitors on ovarian function. MR analysis used summary statistics from large European-ancestry genome-wide association studies (GWAS), applying the inverse-variance weighted (IVW) method, supported by sensitivity analyses. For in vitro experiments, mouse follicles were cultured with 10 µg/ml PD-1 inhibitor ch15mt (clinically relevant concentration), 200 nM doxorubicin (DOX), or PBS control. Follicular morphology was evaluated via diameter measurements; endocrine function by estradiol (E2) quantification using ELISA. Real-time cytoplasmic Ca²⁺ dynamics were monitored using FRET-based Cyto-Ca2 + probes for high-resolution stress assessment. MR results showed no significant association between genetically predicted PD-1 levels and risks of premature ovarian insufficiency, infertility, or alterations in ovarian hormones including AMH and E2. Sensitivity analyses confirmed MR robustness. In vitro, PD-1 inhibition did not affect follicular size or E2 secretion. Notably, DOX induced rapid Ca²⁺ elevation, while PD-1 inhibitor treatment had no detectable effect on Ca²⁺ fluctuations. This first integrative MR and experimental study demonstrates that PD-1 inhibitors at clinically relevant concentrations lack acute ovarian toxicity. While further work is needed to assess long-term effects, these findings demonstrate that standard anti-PD-1 immunotherapy regimens do not compromise follicular viability or endocrine function, strongly supporting their safety in fertility-sparing oncology protocols.</div></div>","PeriodicalId":21137,"journal":{"name":"Reproductive toxicology","volume":"138 ","pages":"Article 109057"},"PeriodicalIF":2.8,"publicationDate":"2025-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145058625","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}