Reproductive biology最新文献

{"title":"Chronic ozone exposure does not alter sexual behavior but modulates oxidative stress and early testicular apoptosis in adult male rats","authors":"Ahmet Yardimci ,&nbsp;Nazife Ulker Ertugrul ,&nbsp;Ramazan Fazil Akkoc ,&nbsp;Nalan Kaya Tektemur ,&nbsp;Ahmet Tektemur ,&nbsp;Elif Erdem Guzel ,&nbsp;Osman Hanbeyoglu ,&nbsp;Ahmet Kavakli ,&nbsp;Murat Ogeturk","doi":"10.1016/j.repbio.2026.101185","DOIUrl":"10.1016/j.repbio.2026.101185","url":null,"abstract":"<div><div>Ozone (O₃) has been used to treat various diseases for many years, with most preclinical studies focusing on its effects in conditions such as testicular torsion and ischemia–reperfusion injury; however, its impact on male reproductive function, particularly sexual behavior, remains largely unexplored. This study aimed to evaluate the effects of chronic O₃ exposure on sexual behavior, reproductive parameters, oxidative stress, and apoptosis in adult male rats. Animals were assigned to a vehicle group (n = 7), which received saline, or an O₃-treated group (n = 7), which received intraperitoneal injections of an O₂/O₃ mixture (1 mL containing 150 µg/kg O₃) three times per week for eight weeks. Behavioral assessments conducted at the end of the treatment period showed that chronic O₃ exposure did not alter appetitive or consummatory sexual behaviors; however, it significantly reduced serum testosterone levels, increased serum total oxidant status (TOS), and decreased testicular hydrogen peroxide (H₂O₂) levels, suggesting a hormetic response. Additionally, O₃ treatment altered apoptotic markers without causing histopathological damage, indicating the onset of early-stage apoptosis. Overall, O₃ exposure did not adversely affect sexual behavior independently of testosterone levels in adult male rats, but its induction of oxidative stress and early apoptosis highlights the need for further studies to clarify underlying mechanisms and establish long-term safety.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 2","pages":"Article 101185"},"PeriodicalIF":2.5,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146145278","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Adipogenesis and myogenesis transcriptome in Piau local breed conceptuses and its comparison with Commercial pig data","authors":"Tânia Fernandes Martins ,&nbsp;Susana Amaral Teixeira ,&nbsp;Lucas Lima Verardo ,&nbsp;Jane de Oliveira Peixoto ,&nbsp;Simone Eliza Facioni Guimarães","doi":"10.1016/j.repbio.2026.101182","DOIUrl":"10.1016/j.repbio.2026.101182","url":null,"abstract":"<div><div>The growth and development of skeletal muscle, along with intramuscular fat deposition, determine the quantity and quality of pork. The Brazilian Piau breed, which has not undergone genetic improvement, exhibits slower growth, greater fat deposition, and increased disease resistance compared to Commercial lines, making it a valuable genetic resource and a relevant model for studying the molecular regulation of myogenesis and adipogenesis. This study aimed to analyze RNA sequencing (RNA-seq) data from Piau pigs to identify differentially expressed genes and biological processes during two stages of intrauterine development, at 25 and 35 days of gestation. Furthermore, gene expression profiles of Piau conceptuses were compared to those of a Commercial line to investigate transcriptional differences between genetic groups. RNA-seq data from seventeen Piau conceptuses (8 embryos and 9 fetuses) were analyzed and compared with public data from a Commercial pig line (Large White × Landrace × Duroc). Differential expression analysis was performed in R, considering FDR &lt; 0.05 and |log2FC| ≥ 2.0, and functional enrichment was evaluated in Cytoscape. In the Piau breed, 43 gene ontology terms showed significant enrichment, mainly related to myogenesis and adipogenesis. In the Commercial breed, 65 enriched terms were identified, predominantly associated with myogenesis. In total, 41 genes showed differential expression in Piau and 55 in Commercial between gestational stages. Key genes associated with adipogenesis and myogenesis were identified, some with functions dependent on the genetic group. These findings contribute to the understanding of molecular differences between genetic groups and support future breeding strategies.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 2","pages":"Article 101182"},"PeriodicalIF":2.5,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146168607","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Investigating trophoblast invasion and angiogenesis expression changes in a caloric deficient mouse model of fetal growth restriction","authors":"James R. Bardill ,&nbsp;Caitlin R. Eason ,&nbsp;Holly Wood ,&nbsp;Courtney Breckenfelder ,&nbsp;Lauren T. Gallagher ,&nbsp;Madison Crew ,&nbsp;Anis Karimpour-Fard ,&nbsp;Carmen C. Sucharov ,&nbsp;Theresa L. Powell ,&nbsp;Clyde J. Wright ,&nbsp;S.Christopher Derderian","doi":"10.1016/j.repbio.2026.101179","DOIUrl":"10.1016/j.repbio.2026.101179","url":null,"abstract":"<div><div>Fetal growth restriction (FGR) is a severe pregnancy complication often caused by placental insufficiency. Proper trophoblast invasion is essential for placental development and function, ensuring adequate nutrient and oxygen supply to the developing fetus. Dysregulation impairs placental perfusion, leading to FGR. This study uses a calorie-restricted mouse model to investigate genes/molecular mechanisms regulating trophoblast invasion across gestational timepoints. Pregnant mice received either a standard or 50 % calorie-restricted diet from E8.5. Placentas and invasion sites were analyzed at E10.5, E12.5, E14.5, E16.5, and E17.5. mRNA sequencing and RT/qPCR examined trophoblast invasion-related genes (<em>Mmp2, Mmp9, Efna1, Rac1, Rras, Ascl2, Tfap2c, Prl7b1</em>) and angiogenesis genes (<em>Vegfa, Vegfb, Pdgf, Akt3</em>). Immunohistochemistry of trophoblast cells (cytokeratin 8, CK8) and endothelial cell markers (endomucin, CD31, CCD105, VEGFR2) was performed. Statistical analysis used Student’s t-test. Caloric restriction significantly reduced fetal/placental weights from E12.5, with persistent growth restriction at E16.5, and E17.5. IHC at E17.5 showed reduced decidual depth, trophoblast invasion distance, and trophoblast quantity within the decidua. This impaired growth was accompanied by reduced expression of trophoblast invasion genes (<em>Mmp2, Mmp9, Efna1, Rac1, Rras, Ascl2, Tfap2c, Prl7b1</em>) in FGR placentas, with a reduction in CK8 trophoblast staining. Angiogenesis reduction in FGR was demonstrated with reduced <em>Vegfa, Vegfb,</em> and <em>Akt3</em> and supported by reduced CD31, CD105, and VEGF2 endothelial cell markers A caloric-restriction mouse model replicates key FGR pathophysiology, including reduced fetal/placental growth, downregulation of trophoblast invasion genes, impaired trophoblast invasion into the decidua, and reduced placenta angiogenesis. These findings offer molecular insights into placental insufficiency that merits further exploration regarding FGR pathogenesis.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 2","pages":"Article 101179"},"PeriodicalIF":2.5,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146038959","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Amygdalin inhibits 17β-estradiol-induced mesenchymal transition through TGF- β/Smad signaling in normal and endometriotic endometrial epithelial cells","authors":"Fang Hu,&nbsp;Jie Zheng,&nbsp;Guoqiong Zhang,&nbsp;Xiaofeng Wen","doi":"10.1016/j.repbio.2026.101184","DOIUrl":"10.1016/j.repbio.2026.101184","url":null,"abstract":"<div><div>Endometriosis is a common gynecological disorder characterized by chronic pelvic pain, infertility, and systemic symptoms, with currently limited treatment options. Amygdalin, a cyanogenic glycoside derived from bitter apricot kernels, has shown potential in endometriosis management, though its molecular mechanism remains unclear. In this study, we investigated the effects of amygdalin on epithelial‑mesenchymal transition (EMT) in both normal and endometriotic endometrial epithelial cells and explored the underlying signaling pathways involved. We found that amygdalin inhibited the proliferation of endometriotic eutopic epithelial cells (EECs) and suppressed 17β‑estradiol‑induced proliferation, invasion, migration, angiogenesis, inflammatory cytokine secretion, and expression of EMT‑related markers in normal epithelial cells (NECs). Mechanistically, amygdalin blocked 17β‑estradiol‑induced EMT in NECs by inhibiting the TGF‑β pathway, as confirmed by TGF‑β knockdown and pharmacological inhibition. Notably, these findings were further validated in primary endometrial epithelial cells isolated from patients with endometriosis, where amygdalin significantly suppressed cell proliferation and downregulated key proteins within the TGF‑β signaling pathway. Taken together, our results demonstrate that amygdalin attenuates 17β‑estradiol‑driven migration, invasion, and EMT in endometrial epithelial cells by modulating TGF‑β signaling. Its efficacy in patient‑derived cells further supports its potential as a therapeutic agent for endometriosis.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 2","pages":"Article 101184"},"PeriodicalIF":2.5,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146189582","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"L-carnitine or melatonin supplementation during vitrification and warming mitigates oxidative stress and improves cryotolerance in immature bovine cumulus-oocyte complexes","authors":"Vivian A.P. Alfradique,&nbsp;Thais de A. Oliveira,&nbsp;Thamiris E.C. Silva,&nbsp;Gicele S. Apolinario,&nbsp;Ribrio Ivan T.P. Batista,&nbsp;Joanna M.G. Souza-Fabjan","doi":"10.1016/j.repbio.2026.101180","DOIUrl":"10.1016/j.repbio.2026.101180","url":null,"abstract":"<div><div>Vitrification of immature cumulus-oocyte complexes (COCs) is a valuable tool in assisted reproductive technologies, but it often induces oxidative stress, reduces viability, and compromises developmental potential. This study investigated the effects of supplementing vitrification and warming solutions with non-enzymatic antioxidants — glutathione (GSH, 5 mM), <span>L</span>-carnitine (LC, 3.03 mM), or melatonin (MLT, 10⁻⁶ mM) — on nuclear maturation, oxidative stress, early apoptosis, cryosurvival, and gene expression. COCs were vitrified and warmed in the presence or absence (negative control) of antioxidants; non-vitrified COCs served as fresh controls. Vitrification significantly reduced (P &lt; 0.05) viability, survival, and nuclear maturation rates compared to fresh COCs. However, MLT increased viability, and LC enhanced nuclear maturation (P &lt; 0.05 vs negative control). Gap junction activity and early apoptosis were not significantly affected by antioxidant treatment. Intracellular GSH and ROS levels were restored in antioxidant-treated groups, comparable to fresh controls (P &gt; 0.05), whereas the negative control had increased ROS and decreased GSH (P &lt; 0.05). In cumulus cells, expression of mitochondrial genes <em>ATP6</em> and <em>ATP8</em>—key for oxidative phosphorylation—was downregulated in the negative control. Moreover, LC and MLT supplementation counteracted this effect, upregulating <em>SOD2</em> (mitochondrial superoxide dismutase) and S<em>OD1</em> (cytosolic isoform), respectively. In oocytes, both LC and MLT upregulated SOD1 (P &lt; 0.05). In summary, these findings suggest that LC and MLT supplementation improve cryotolerance and mitigate oxidative stress in vitrified immature bovine COCs by modulating antioxidant defenses and mitochondrial gene expression.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 2","pages":"Article 101180"},"PeriodicalIF":2.5,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145980662","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metabolic–endocrine remodelling of the testis under polystyrene nanoplastic exposure: Intervention by organ-specific phytocomplexes of Nelumbo nucifera","authors":"Putri Ayu Ika Setiyowati ,&nbsp;Febriyansyah Saputra ,&nbsp;Firli Rahmah Primula Dewi ,&nbsp;Mochammad Aqila Herdiansyah ,&nbsp;Vuanghao Lim ,&nbsp;Alfiah Hayati","doi":"10.1016/j.repbio.2026.101183","DOIUrl":"10.1016/j.repbio.2026.101183","url":null,"abstract":"<div><div>Polystyrene nanoplastics (PS-NPs) are emerging endocrine-disrupting pollutants that can accumulate systemically and impair male reproduction by inducing oxidative stress, hormonal dysregulation, and disrupted spermatogenesis. <em>Nelumbo nucifera</em> (lotus) is a medicinal aquatic plant rich in polyphenols and flavonoids with antioxidant and anti-inflammatory properties. This study evaluated the protective effects of <em>N. nucifera</em> leaf, flower, and rhizome extracts against PS-NP-induced testicular dysfunction in <em>Rattus norvegicus</em>. Male rats were orally exposed to PS-NPs (200 µg/kg bw) and co-treated with <em>N. nucifera</em> extracts (200 mg/kg bw) for 55 days, quercetin an effective free-radical scavenger and anti-inflammatory, was utilized as a reference. PS-NP exposure resulted in testicular accumulation and pronounced oxidative and inflammatory injury, evidenced by increased malondialdehyde (MDA) and elevated pro-inflammatory cytokines (IL-1β, IL-6, TNF-α) alongside reduced antioxidant enzyme activities (SOD, CAT). PS-NPs also perturbed testicular energy metabolism (GLUT1, HK1, PKM, GYS1, GLUT8), suppressed steroidogenic genes (HSD3β, HSD17β), reduced serum luteinizing hormone (LH), and impaired sperm quality. Co-treatment with <em>N. nucifera</em> extracts mitigated these alterations, with the rhizome extract demonstrating the most significant enhancement. This tendency aligned with the rhizome's elevated total phenolic content and ABTS radical-scavenging activity, corresponding with a reduced testicular PS-NP accumulation. Molecular docking further supported mechanistic relevance, showing high predicted affinities of lanosterol and cycloartenol toward SOD, CAT, LH receptor (LHR), and androgen receptors. These findings demonstrate that <em>N. nucifera</em>, especially the rhizome exerts potent antioxidant, anti-inflammatory, and endocrine-restorative effects against nanoplastic-induced testicular toxicity, highlighting its potential as a multitarget phytotherapeutic agent.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 2","pages":"Article 101183"},"PeriodicalIF":2.5,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146145373","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"HDAC2 regulates oocyte maturation in Tan sheep via its strong association with regulating H4K12 acetylation and EREG secretion","authors":"Junli He ,&nbsp;Yaxiu Xu ,&nbsp;Xiangyan Wang ,&nbsp;Zhipeng Qi ,&nbsp;Donghuan Lv ,&nbsp;Hongyuan Song ,&nbsp;Yujie Yan ,&nbsp;Jinghua Wang ,&nbsp;Sirui Min ,&nbsp;Guoliang Xia ,&nbsp;Qiang Chen ,&nbsp;Xinfeng Liu","doi":"10.1016/j.repbio.2026.101186","DOIUrl":"10.1016/j.repbio.2026.101186","url":null,"abstract":"<div><div>This study aimed to explore the potential mechanism of histone deacetylase 2 (HDAC2) in follicular development and oocyte maturation of Tan sheep, and clarify whether it affects oocyte maturation by responding to luteinizing hormone (LH) signals, regulating histone acetylation, and promoting epidermal growth factor (EGF)-like factor secretion. The effect of HDAC2 inhibitor on oocyte maturation rate was evaluated via the Cumulus-oocyte complex (COC) and granulosa cell co-culture system. Related indicators were detected using immunohistochemistry, immunofluorescence, reverse transcription-quantitative polymerase chain reaction (RT-qPCR), Western blotting, and enzyme-linked immunosorbent assay (ELISA). Results showed that HDAC2 was localized in the nuclei of granulosa cells and oocytes in all stages of Tan sheep follicles, with its expression significantly upregulated during follicular development (<em>P &lt; 0.001</em>). RT-qPCR and Western blotting results confirmed that FSH significantly upregulated HDAC2 mRNA and protein expression in granulosa cells (<em>P &lt; 0.05</em> or <em>P &lt; 0.01</em>), while LH significantly downregulated HDAC2 mRNA and protein levels after 2-hour treatment (<em>P &lt; 0.01</em> or <em>P &lt; 0.0001</em>). Treatment with 20 μM CAY10683, a specific HDAC2 inhibitor, significantly increased the acetylation level of histone H4 lysine 12 (H4K12ac) in granulosa cells (<em>P &lt; 0.001</em>), and specifically promoted the secretion and expression of epiregulin (EREG) in granulosa cells at both mRNA and protein levels (<em>P &lt; 0.01</em> or <em>P &lt; 0.001</em>). Additionally, CAY10683 enhanced the in vitro maturation rate of oocytes. In conclusion, HDAC2 affects Tan sheep oocyte maturation by responding to LH signals, and the results suggest a potential link between H4K12 acetylation and EREG secretion in this process, providing a basis for studying the epigenetic regulation of sheep oocytes.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 2","pages":"Article 101186"},"PeriodicalIF":2.5,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146184041","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Decreased autophagic activity in spermatozoa from infertile men adversely affects early embryonic development","authors":"Mehmet Alper Arslan ,&nbsp;Aysın Pınar Türkmen ,&nbsp;Ramazan Aşcı","doi":"10.1016/j.repbio.2025.101177","DOIUrl":"10.1016/j.repbio.2025.101177","url":null,"abstract":"<div><div>Oligoasthenoteratozoospermia (OAT) is a complex reproductive health problem characterized by impaired sperm parameters. Autophagy serves critical functions in diverse stages of male reproduction and has been shown to be functionally active in mature ejaculate spermatozoa from healthy men. The present study aims to investigate the level of autophagic activity in spermatozoa from OAT patients and its association with embryo development following intracytoplasmic sperm injection (ICSI). 42 OAT men with healthy spouses and 21 age-matched fertile controls were included in the study. Following semen analysis, semen samples were processed for sperm protein isolation and protein concentrations were measured by BCA assay. Autophagic activity levels in spermatozoa, i.e. LC3 conversion, p62 and Beclin-1 protein levels, were measured by Western blotting. In vitro fertilization was performed by routine ICSI. Fertilization and embryo development rates were calculated accordingly. Spermatozoal autophagic activity was significantly decreased in OAT patients compared to fertile controls, as demonstrated by concurrently decreased LC3-II and increased p62 protein levels. Beclin-1 levels did not significantly change between the two groups. Evaluation of p62 correlation and LC3-II regression analyses with ICSI success rates revealed that low spermatozoal autophagic activity is significantly associated with a decline in day 3 embryo development. Our findings reveal an adverse late paternal effect on early embryonic development, which suggests an involvement of sperm DNA damage as a consequence of decreased sperm autophagy. Understanding the interplay between sperm autophagy and embryo quality will help develop novel therapeutic strategies to increase sperm autophagic activity in OAT patients in the future.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 2","pages":"Article 101177"},"PeriodicalIF":2.5,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145908771","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Hexavalent chromium inhibits testosterone synthesis in bovine testicular leydig cells through mitochondrial damage via BNIP3","authors":"Hongxia Li ,&nbsp;Jia Liu ,&nbsp;Liwei Huang ,&nbsp;Yu Cheng ,&nbsp;Guoqing Zhao ,&nbsp;Xiaolong Pan ,&nbsp;Lijuan Huang ,&nbsp;Pengkang Song ,&nbsp;Le Zhao ,&nbsp;Xuanqi Yu ,&nbsp;Juan Xiong ,&nbsp;Qing Tian ,&nbsp;Xi Wang ,&nbsp;Xiaoyu Li ,&nbsp;Ruigao Song","doi":"10.1016/j.repbio.2026.101181","DOIUrl":"10.1016/j.repbio.2026.101181","url":null,"abstract":"<div><div>This study investigated the molecular mechanisms underlying the inhibitory effects of hexavalent chromium (Cr(Ⅵ)) exposure on testosterone synthesis in bovine testicular Leydig cells. First, bovine Leydig cells were isolated and purified, cultured after purity identification, and the effect of different dose gradients of Cr(Ⅵ) on cell viability after 12 h of exposure was detected using the CCK-8 method. Cell proliferation was assessed using a cell proliferation assay kit. Testosterone levels were measured by ELISA. Intracellular reactive oxygen species (ROS) levels were detected using the DCFH-DA fluorescent probe. Cell autophagy was examined by the monodansylcadaverine (MDC) method. Changes in mitochondrial membrane potential were determined using the JC-1 probe. Ultrastructural changes in mitochondria were observed via transmission electron microscopy. The expression of key genes involved in testosterone synthesis, cell autophagy, mitochondrial fusion, mitochondrial fission, and mitophagy was detected by qRT-PCR. Differentially expressed genes were screened through RNA-seq. Our results demonstrate that Cr(Ⅵ) treatment significantly suppressed testosterone synthesis. Concurrently, Cr(Ⅵ) enhanced intracellular MDC levels and induced mitochondrial dysfunction, characterized by ultrastructural damage, reduced mitochondrial membrane potential, and disrupted mitochondrial dynamics. RNA sequencing analysis revealed that Cr(Ⅵ) activates <em>BNIP</em>3 recruitment and high expression within cells. Subsequent <em>BNIP</em>3 silencing using siRNA interference significantly attenuated Cr(Ⅵ)-induced abnormalities in the expression of mitochondrial genes and intracellular ROS accumulation. Moreover, <em>BNIP</em>3 expression knockdown markedly upregulated the mRNA expression of key steroidogenic genes and the level of testosterone in cell supernatant. These findings suggest that Cr(Ⅵ) inhibits testosterone synthesis in Leydig cells through <em>BNIP</em>3-mediated mitochondrial damage. This study identifies <em>BNIP</em>3 as a potential therapeutic target and provides a theoretical foundation for addressing Cr(Ⅵ)-associated male reproductive dysfunction.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 2","pages":"Article 101181"},"PeriodicalIF":2.5,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146038520","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Branched chain amino acid sufficiency is necessary for proper luteinizing hormone response and testosterone synthesis","authors":"Yeva Shamailova ,&nbsp;Saad A. Farooq ,&nbsp;Megan E. Gilmore ,&nbsp;Timothy J. Stanek ,&nbsp;Esther M. Lopez ,&nbsp;Berish B. Wetstein ,&nbsp;Emily T. Mirek ,&nbsp;Tracy G. Anthony ,&nbsp;Elizabeth M. Snyder","doi":"10.1016/j.repbio.2025.101094","DOIUrl":"10.1016/j.repbio.2025.101094","url":null,"abstract":"<div><div>Testosterone production by testicular Leydig cells (steroidogenesis) is vital to male fertility and overall male health. Information about how nutrition influences Leydig cell steroidogenesis is lacking. Branched chain amino acids (BCAAs – leucine, isoleucine, and valine) are essential amino acids and important regulators of protein synthesis and energy production. Circulating and tissue BCAA levels are tightly regulated by the enzyme branched chain a-keto acid dehydrogenase kinase (BCKDK), which inhibits their catabolism. This work explored how BCAAs, and especially leucine, modulate male fertility and testosterone production. In a mutant mouse model of <em>Bckdk</em>, breeding analysis showed reduced male fertility and circulating testosterone. Further, morphological evaluation demonstrated testicular and epididymal abnormalities consistent with abnormal testicular androgen signaling. Fertility was partially rescued by feeding a high protein diet while circulating testosterone was not. In wild type testes, Leydig cells were the primary cell type to express BCKDK. Leveraging a primary interstitial cell culture, cell survival and apoptosis analyses demonstrated Leydig cells are highly sensitive to leucine deprivation and this sensitivity is enhanced under steroidogenesis stimulating conditions. Lastly, using the same primary cell culture system, testosterone production was shown to be lost under leucine deprivation. In total, this work demonstrates Leydig cells are uniquely sensitive to BCAA status under steroidogenesis stimulation and that regulated BCAA catabolism may be important for optimal male fertility.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 1","pages":"Article 101094"},"PeriodicalIF":2.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145418812","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}