Reproductive toxicology最新文献

{"title":"New approach methodologies for assessing developmental toxicity of pharmaceuticals: Case examples and future directions","authors":"Nicola Powles-Glover ,&nbsp;Allen R. Kaczor ,&nbsp;Steven Van Cruchten ,&nbsp;Christopher J. Bowman ,&nbsp;Kimberly C. Brannen ,&nbsp;Claudia Demarta-Gatsi ,&nbsp;Isabelle LeConte ,&nbsp;Amer Jamalpoor ,&nbsp;Fumito Mikashima ,&nbsp;Shermaine Mitchell-Ryan ,&nbsp;Dinesh Stanislaus ,&nbsp;Peter Theunissen ,&nbsp;Belen Tornesi ,&nbsp;Ronald L. Wange ,&nbsp;Connie L. Chen","doi":"10.1016/j.reprotox.2025.109035","DOIUrl":"10.1016/j.reprotox.2025.109035","url":null,"abstract":"<div><div>Several new approach methodologies (NAMs) for developmental toxicity (Dev Tox) testing are being used by pharmaceutical companies for derisking or for exploring Dev Tox mechanisms. Regulatory adoption of these NAMs-based approaches as being adequate for Dev Tox risk assessment has been more challenging, due, in part, to dynamic changes in the conceptus and placenta throughout development and the impact of the pharmaceutical on the mother’s physiology, which may also have an embryo-fetal impact. Still, there is currently a recognition by Health Authorities that there are certain contexts-of-use under which Dev Tox NAMs can provide information that is adequate to inform risk. This has been adopted in the 3rd revision of the ICH S5 guideline, which provides a path to qualify Dev Tox NAMs for regulatory decision making. Despite this opportunity, pharmaceutical companies rarely submit Dev Tox NAMs to Health Authorities for qualification or with intent to support regulatory decision making. This may be in part due to the need for a greater understanding of the biological relationship between currently available Dev Tox NAMs and in vivo outcome, applicability domain, translatability, predictivity, and that these NAMs do not cover the complete scope of embryo-fetal development. Furthermore, there is a lack of Dev Tox NAMs data visibility to Health Authorities. To use Dev Tox NAMs for regulatory decision making, more data sharing with Health Authorities and further understanding the applicability domain of these methodologies are needed.</div></div>","PeriodicalId":21137,"journal":{"name":"Reproductive toxicology","volume":"137 ","pages":"Article 109035"},"PeriodicalIF":2.8,"publicationDate":"2025-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144966862","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":"Chikusetsusaponin IVa ameliorates paroxetine-induced Leydig cells (TM3 cells) injury via the Nrf2/HO-1 signaling pathway","authors":"Qianqian Huang ,&nbsp;Haiying Wu ,&nbsp;Xiangxin Xiao ,&nbsp;Zhen Lu ,&nbsp;Xiuping Fan ,&nbsp;Wenhong Cao ,&nbsp;Suqing Liu ,&nbsp;Xiaoming Qin","doi":"10.1016/j.reprotox.2025.109039","DOIUrl":"10.1016/j.reprotox.2025.109039","url":null,"abstract":"<div><div>Paroxetine (PRX) exhibits significant toxic effects on the male reproductive system. Previous animal studies have demonstrated that <em>Pfaffia glomerata</em> extract can ameliorate PRX-induced sexual dysfunction in male mice, but its active components and underlying mechanisms remain unclear. Chikusetsusaponin IVa (CHS-IVa), a major saponin component of <em>Pfaffia glomerata</em> with well-documented antioxidant and anti-apoptotic activities, has become a research focus. This study aimed to investigate whether CHS-IVa could mitigate PRX-induced injury in mouse Leydig cells (TM3 cells) by regulating oxidative stress, apoptosis, and androgen synthesis pathways. To achieve this, a PRX-induced injury model in TM3 cells was established and subjected to comprehensive evaluation using CCK-8 assay for cell viability, ELISA for sex hormone levels, DCFH-DA fluorescent probe for reactive oxygen species (ROS) detection, and RT-qPCR/Western blot for mRNA and protein expression analysis. Results showed that compared to PRX group, CHS-IVa (6.25 μg/mL) significantly increased cell viability by 12.9 % (<em>p</em> &lt; 0.05); activated Nrf2/HO1 signaling pathway, reducing intracellular ROS levels by at least 21.9 % (<em>p</em> &lt; 0.05), thereby alleviating oxidative stress injury; upregulated mRNA expression of androgen synthesis-related genes (StAR, CYP11a1, CYP17a1, LHr) by over 2-fold (<em>p</em> &lt; 0.05), with maximal increases in testosterone, dihydrotestosterone and luteinizing hormone levels by 8.4 %, 50.4 % and 13.0 % respectively (<em>p</em> &lt; 0.05); enhanced anti-apoptotic factor Bcl-2 mRNA and protein expression by up to 3.4-fold and 1.6-fold (<em>p</em> &lt; 0.05), while reducing pro-apoptotic factor Bax mRNA and protein expression by 40.5 % and 44.6 % (<em>p</em> &lt; 0.05), and decreasing Caspase-3 mRNA expression by 61.5 % (<em>p</em> &lt; 0.05), ultimately reducing PRX-induced abnormal apoptosis in TM3 cells. In conclusion, CHS-IVa serves as the key active component in <em>Pfaffia glomerata</em> that protects against PRX-induced reproductive toxicity by ameliorating injury in TM3 cells through multiple mechanisms.</div></div>","PeriodicalId":21137,"journal":{"name":"Reproductive toxicology","volume":"137 ","pages":"Article 109039"},"PeriodicalIF":2.8,"publicationDate":"2025-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144916715","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":"Abamectin disrupts sperm function through the alteration of PKA activity and tyrosine phosphorylation in boar spermatozoa","authors":"Claudine Uwamahoro ,&nbsp;Jae-Hwan Jo ,&nbsp;Seung-Ik Jang ,&nbsp;Eun-Ju Jung ,&nbsp;Woo-Jin Lee ,&nbsp;Jeong-Won Bae ,&nbsp;Daehyun Kim ,&nbsp;Junkoo Yi ,&nbsp;Sangsu Shin ,&nbsp;Joonho Moon ,&nbsp;Woo-Sung Kwon","doi":"10.1016/j.reprotox.2025.109040","DOIUrl":"10.1016/j.reprotox.2025.109040","url":null,"abstract":"<div><div>The potential toxicity of abamectin (ABM), a macrocyclic lactone insecticide widely used in agriculture, in non-target organisms has raised concerns. Therefore, this study aimed to investigate the effects of ABM on sperm function, focusing on motility, kinematics, ATP levels, viability, protein kinase A (PKA) activity, and tyrosine phosphorylation. Boar spermatozoa were incubated with various ABM concentrations (0, 1, 5, 10, 15, 25, 50, and 100 µM) under capacitating conditions and assessed through computer-assisted sperm analysis, Hoechst 33258/chlortetracycline staining, ATP and viability assays, and Western blotting. The results revealed a dose-dependent decline in sperm motility and ATP levels starting at 10 µM. Sperm capacitation was significantly altered at concentrations of at least 25 µM, with an increase in capacitated spermatozoa and a decrease in non-capacitated spermatozoa. However, overall cell viability remained unaffected. Western blot analysis showed abnormal alterations in PKA substrates and tyrosine-phosphorylated proteins. These findings suggest ABM disrupts sperm function and key fertilization pathways, indicating potential risks to male fertility.</div></div>","PeriodicalId":21137,"journal":{"name":"Reproductive toxicology","volume":"137 ","pages":"Article 109040"},"PeriodicalIF":2.8,"publicationDate":"2025-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144925166","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 herbicide mixtures on the fertilizing capacity of bovine sperm","authors":"Diogo Ferreira Bicca ,&nbsp;Rafaela Dalmolin Menezes ,&nbsp;Laura Rohde Brondani ,&nbsp;Monike Quirino ,&nbsp;Larissa Thaísa Weide ,&nbsp;Fábio Gallas Leivas ,&nbsp;Daniela dos Santos Brum ,&nbsp;Francielli Weber Santos Cibin","doi":"10.1016/j.reprotox.2025.109037","DOIUrl":"10.1016/j.reprotox.2025.109037","url":null,"abstract":"<div><div>Herbicides are the most commonly used pesticide type worldwide. In Brazil, glyphosate-, dichlorophenoxyacetic acid (2,4-D)-, and atrazine-based pesticide formulations are intensively applied to crops, and mixtures of these compounds occur frequently in the environment. Owing to their proximity to these areas and management practices, bovines are exposed to these pesticide mixtures, and their impact on their health is unknown. In this study, we evaluated the <em>in vitro</em> effects of herbicide mixtures on bovine sperm. Semen from four bulls was prepared as a pool and divided into groups: control, dimethyl sulfoxide (DMSO), glyphosate (Gly; 50 µg/mL), 2,4-D (0.11 µg/mL), atrazine (Atz; 0.0107 µg/mL), Gly + 2,4-D (GD), Gly + Atz (GA), Atz+ 2,4-D (AD) and Gly + 2,4-D + Atz (GDA). Sperm cells were evaluated after 3 h of incubation with the different treatments at 37℃. Results showed that the Gly, 2,4-D, and AD groups decreased progressive motility, mean path velocity, and beat cross frequency compared to the control group. Similarly, Gly and 2,4-D reduced curvilinear and straight-line velocities, and 2,4-D affected the amplitude of lateral head displacement. Although no differences in reactive species levels were detected, an overall reduction in antioxidant capacity was observed. Membrane integrity, acrosome damage, and mitochondrial membrane potential results did not differ significantly among the groups. However, the mixture diminished sperm fertilization capacity in groups GD, AD, and GDA, when compared to the control. No effects appeared in the DMSO group. Herbicides showed distinct impacts on bovine sperm, emphasizing the importance of evaluating pesticide mixtures thoroughly.</div></div>","PeriodicalId":21137,"journal":{"name":"Reproductive toxicology","volume":"137 ","pages":"Article 109037"},"PeriodicalIF":2.8,"publicationDate":"2025-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144896738","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":"Eugenol mitigates glyphosate-induced testicular toxicity via modulation of oxidative stress, ER stress, RAGE/NLRP3 and PI3K/AKT signaling pathways in rats","authors":"Tuba Dogan ,&nbsp;Ismail Bolat ,&nbsp;Samet Tekin ,&nbsp;Burak Cınar ,&nbsp;Esra Aktas Senocak ,&nbsp;Omercan Alat ,&nbsp;Mesut Bünyami Halici","doi":"10.1016/j.reprotox.2025.109038","DOIUrl":"10.1016/j.reprotox.2025.109038","url":null,"abstract":"<div><div>Glyphosate (GLY), a widely used herbicide, has been implicated in male reproductive toxicity through mechanisms involving oxidative stress, apoptosis, and disrupted cellular signaling. This study evaluated the protective effects of eugenol (EU), a natural antioxidant, against GLY-induced testicular damage in rats. Thirty-five adult male rats were divided into five groups: Control, EU (100 mg/kg), GLY (150 mg/kg), GLY+EU (50 mg/kg), and GLY+EU (100 mg/kg). Treatments were administered orally for seven days. Testicular tissue was analyzed histologically (H&amp;E staining), and PI3K/AKT signaling was assessed via immunohistochemistry and immunofluorescence. Oxidative stress markers (MDA, GSH, SOD, CAT, GPx) were measured biochemically. Gene expressions related to ER stress, RAGE, and NLRP3 were evaluated by qRT-PCR, while apoptotic (Bax, Bcl-2, Caspase-3), inflammatory (Beclin-1, NF-κB, TNF-α), and antioxidant (Keap1, Nrf2) protein levels were analyzed by western blotting. Co-treatment with EU, especially at 100 mg/kg, significantly ameliorated GLY-induced testicular toxicity by reducing oxidative and ER stress, inhibiting inflammation and apoptosis, and modulating the PI3K/AKT pathway. These findings highlight the potential of eugenol as a protective agent against GLY-induced reproductive toxicity through its regulatory effects on multiple molecular pathways.</div></div>","PeriodicalId":21137,"journal":{"name":"Reproductive toxicology","volume":"137 ","pages":"Article 109038"},"PeriodicalIF":2.8,"publicationDate":"2025-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144889614","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":"Estrogen receptor 1 (ESR1) promotes Di-butyl phthalate (DBP)-induced hypospadias by regulating mitophagy through PINK1-Parkin axis to inhibit ferroptosis","authors":"Jinping Liu ,&nbsp;Yi Cheng ,&nbsp;Enfu Huang ,&nbsp;Zhixiang Liu ,&nbsp;Yun Zhou","doi":"10.1016/j.reprotox.2025.109036","DOIUrl":"10.1016/j.reprotox.2025.109036","url":null,"abstract":"<div><div>This study reveals how Di-butyl phthalate (DBP), an estrogen-mimicking environmental pollutant, induces hypospadias by inhibiting ferroptosis through ESR1 activation and PINK1-Parkin-dependent mitophagy. Utilizing a prenatal DBP-exposed fetal rat hypospadias model, we observed significant downregulation of pro-ferroptotic ACSL4 and upregulation of anti-ferroptotic GPX4/SLC7A11 in urethral tissues, alongside elevated oxidative stress markers (MDA, Fe²⁺) and reduced glutathione (GSH). In vitro experiments using rat urethral plate fibroblasts (RUPFs) demonstrated that DBP enhanced ferroptosis resistance and promoted proliferation at concentrations below 200 μM. Mechanistically, DBP activated ESR1, which triggered mitophagy via the PINK1-Parkin pathway, reducing mitochondrial damage and reactive oxygen species (ROS) accumulation, thereby suppressing ferroptosis. Inhibition or silencing of ESR1 reversed these effects, restoring ferroptosis sensitivity and oxidative stress. These findings unveil a novel ESR1-mitophagy-ferroptosis regulatory axis, linking DBP exposure to hypospadias pathogenesis. The study not only elucidates a previously unrecognized molecular pathway underlying phthalate-induced congenital malformations but also identifies ESR1 and mitophagy as potential therapeutic targets. By integrating in vivo and in vitro approaches, this work advances the understanding of environmental endocrine disruptors’ role in developmental toxicity and provides actionable insights for mitigating their health impacts, aligning with current priorities in reproductive and environmental health research.</div></div>","PeriodicalId":21137,"journal":{"name":"Reproductive toxicology","volume":"137 ","pages":"Article 109036"},"PeriodicalIF":2.8,"publicationDate":"2025-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144908971","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":"Study on the mechanism of different doses of lycium barbarum polysaccharides affecting sperm motility in Drosophila melanogaster","authors":"Jingjing Yu ,&nbsp;Wenjun Fu ,&nbsp;Xinyi Wang ,&nbsp;Xiang Zhang ,&nbsp;Kun Xiong ,&nbsp;Yuan Wang","doi":"10.1016/j.reprotox.2025.109034","DOIUrl":"10.1016/j.reprotox.2025.109034","url":null,"abstract":"<div><div>Lycium barbarum, a Solanaceae plant, yields bioactive polysaccharides (LBP) in its dried mature fruits. LBP has demonstrated protective effects on the reproductive system. This study took <em>Drosophila melanogaster</em> as the experimental subject to investigate the dose-dependence of LBP on male reproductive capacity. The flies were orally administered LBP (0–2000 mg/L) by the CAFE method. Twenty-four hours later, sperm motility, ovulation rate and testicular gene expression were evaluated. Key results: LBP modulates sperm motility and female ovulation rate in a dose-dependent manner. The dose-response curves of <em>oamb, gish, zeste</em> and <em>cg9465</em> show an inverted U-shaped trend. LBP regulates spermatogenic genes through OAMB receptor. The PKA signal was positively correlated with histone methyltransferase (<em>zeste</em>), indicating that there are other regulatory pathways. Two-phase dose effects were observed: LBP enhanced motility at 0–150 mg/kg and 450–1000 mg/kg, while inhibited motility at 150–450 mg/kg and &gt; 1000 mg/kg. In conclusion, the reproductive effects of LBP follow nonlinear dynamic laws. This complexity further highlights the necessity of conducting in-depth studies on each component of LBP in order to accurately assess its reproductive toxicity and optimize its therapeutic potential.</div></div>","PeriodicalId":21137,"journal":{"name":"Reproductive toxicology","volume":"137 ","pages":"Article 109034"},"PeriodicalIF":2.8,"publicationDate":"2025-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144864238","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":"Beauvericin disrupts G2/M transition and induces meiotic arrest during mouse oocyte maturation","authors":"Yejin Kim ,&nbsp;Yu-Jin Jo ,&nbsp;Seung-Bin Yoon ,&nbsp;Jeongwoo Kwon ,&nbsp;Hyeong-Ju You ,&nbsp;Changsic Youn ,&nbsp;Young-Kug Choo ,&nbsp;Ji-Su Kim","doi":"10.1016/j.reprotox.2025.109032","DOIUrl":"10.1016/j.reprotox.2025.109032","url":null,"abstract":"<div><div>Beauvericin (BEA) is a mycotoxin produced by fungi of the genus <em>Fusarium</em> that causes adverse toxic effects in humans and livestock. Previous studies have demonstrated that BEA causes reproductive toxicity in pigs and juvenile sheep. However, the effects of BEA on meiotic resumption and the underlying mechanisms remain unclear. In this study, we investigated the molecular mechanisms underlying meiotic failure caused by the toxic effects of BEA on fully grown immature mouse oocytes. Exposure to BEA led to DNA damage, affected phosphatidylinositol 3-kinase/protein kinase B/phosphodiesterase 3 A (PI3K/AKT/PDE3A)-mediated cAMP signaling, and inhibited cyclin-dependent kinase (CDK) complex (MPF) activation by modulating CDK1 activity through altered expression of Wee1 and CDC25B. These disruptions ultimately led to germinal vesicle arrest during <em>in vitro</em> oocyte maturation. Our findings suggest that BEA treatment blocks germinal vesicle breakdown by affecting the PI3K/AKT/PDE3A-mediated cAMP-MPF pathway, resulting in the failure of meiotic progression and defects in the maturation of mammalian oocytes.</div></div>","PeriodicalId":21137,"journal":{"name":"Reproductive toxicology","volume":"137 ","pages":"Article 109032"},"PeriodicalIF":2.8,"publicationDate":"2025-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144966937","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":"Role of adipose-derived stem cell exosomes in paclitaxel-induced acute ovarian injury: An experimental approach","authors":"Betul Yalcın ,&nbsp;Tugce K. Kalkan ,&nbsp;Zeynep B. Gonen ,&nbsp;Eda Koseoglu ,&nbsp;Gozde O. Onder ,&nbsp;Nur S. Gokdemir ,&nbsp;Munevver Baran ,&nbsp;Arzu Yay","doi":"10.1016/j.reprotox.2025.109033","DOIUrl":"10.1016/j.reprotox.2025.109033","url":null,"abstract":"<div><div>Paclitaxel (PTL) is commonly used in cancer therapy at varying doses and durations, often in combination with other chemotherapeutic agents. However, achieving therapeutic efficacy typically requires high doses, which are associated with considerable toxicity. Adipose-derived stem cells have shown therapeutic potential, particularly through the release of extracellular vesicles known as exosomes. This study investigated the potential protective effects of exosomes derived from adipose-derived mesenchymal stem cells (AMSC-Exos) in a rat model of PTL-induced acute ovarian injury. Twenty-eight rats were assigned to groups: control, PTL (7.5 mg/kg), AMSC-Exos (1 ×10⁶ exosomes), and PTL+AMSC-Exos (7.5 mg/kg PTL + 1 × 10⁶ exosomes). Three days after the administration, ovarian tissues were harvested for histological and biochemical analysis. Hematoxylin and eosin (H&amp;E) and Masson's Trichrome (MT) staining revealed significant histopathological deterioration in the cortex and medulla of ovarian tissue in the PTL group compared to the PTL+AMSC-Exos group. Exosome treatment following PTL administration resulted in upregulation of VEGF and downregulation of HIF-1α, NF_KB-p65, and IL-1β immunostaining intensities. Additionally, AMH immunostaining intensity was increased in primary, preantral, and secondary follicles. Levels of TNF-α, IL-1β, and IL-6 were significantly lower in the exosome treated group than in the PTL group, according to the results of the ELISA. These findings demonstrate that AMSC-Exos exhibited beneficial effects against PTL-induced acute ovarian damage by reducing histopathological alterations, inflammation, and HIF-1α expression, while enhancing VEGF expression and ovarian reserve. AMSC-Exos may represent a promising therapeutic approach for preventing chemotherapy-induced ovarian toxicity.</div></div>","PeriodicalId":21137,"journal":{"name":"Reproductive toxicology","volume":"137 ","pages":"Article 109033"},"PeriodicalIF":2.8,"publicationDate":"2025-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144880143","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":"All-trans Retinoic Acid regulates cellular senescence of mouse embryonic palatal mesenchyme (MEPM) cells in developing cleft palates","authors":"Yang Ma ,&nbsp;Shiyi Huang ,&nbsp;Yingshi Liang ,&nbsp;Yuebin Zhu ,&nbsp;Haofeng Huang ,&nbsp;Bingna Zhang ,&nbsp;Lungang Shi","doi":"10.1016/j.reprotox.2025.109030","DOIUrl":"10.1016/j.reprotox.2025.109030","url":null,"abstract":"<div><h3>Background</h3><div>Non-syndromic cleft lip and palate is one of the most common congenital craniofacial malformations, however, its mechanism is not well understood. A number of relevant studies have confirmed that the cleft lip and palate is caused by the interaction of environmental and genetic factors. Retinoic acid is one of the metabolites of vitamin A and is involved in various physiological functions in the body, which is essential for the regulation of cell growth and differentiation and the maintenance of normal human development. However, excessive intake of All-trans Retinoic Acid (atRA) is one of the etiological factors contributing to the development of cleft palate. Cell senescence is a hot topic and a new direction in recent years, which is related to the occurrence of various tumors and diseases. From the perspective of cellular senescence, this study aimed to verify the mechanism of action of atRA-induced cleft palate in model of mouse embryonic palatal mesenchyme (MEPM) cells. This study aims to verify whether the pathogenesis of the atRA-induced cleft palate occurs because the cellular senescence of MEPM cells via 53/p21 signaling pathway.</div></div><div><h3>Methods</h3><div>The palate tissues of atRA-induced cleft palate were obtained for section staining and western blotting test. MEPM cells were obtained from palate tissues and cultured in vitro, and then MEPM cells in vitro were treated with atRA. To verify that the atRA could affect the cell proliferative ability and cellular senescence of MEPM cells through the p53/p21 pathway, which resulted in the failure of palatal fusion in embryonic mice, leading to cleft palate. The senescence-associated β-Galactosidase staining, CCK-8 activity assay, cell cycle analysis and western blotting test were used.</div></div><div><h3>Results</h3><div>In atRA-induced cleft palate group, tissue sections of the palate showed that there was no change in TUNEL apoptosis fluorescence staining. However, there was increased cellular senescence in MEPM cells treated with atRA as characterized by enhancing senescence-associated β-galactosidase (SA-β-Gal) activity, reducing cell proliferation, inducing MEPM cells cell cycle arrest at G1 phase and increasing expression of the senescence markers p53 and p21. p53/p21 signaling pathway was up-regulated, which could induce the cells to undergo senescence, resulting in a decrease of cell proliferative ability.</div></div><div><h3>Conclusions</h3><div>Our experimental results have shown that atRA could increase cell senescence through the p53/p21 signaling pathway in MEPM cells and diminish cell activity and proliferation ability, inducing the occurrence of cellular senescence and resulting in cleft palate in the embryonic mouse. From the viewpoint of cellular senescence, this study is intended to expand the mechanism of action of atRA-induced cleft palate as well as to provide new ideas for the study of the etiology of cleft palate.</div></div>","PeriodicalId":21137,"journal":{"name":"Reproductive toxicology","volume":"137 ","pages":"Article 109030"},"PeriodicalIF":2.8,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144859564","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}