F&S science最新文献_第4页

The dual role of interleukin-22: balancing protection and risk in pregnancy during inflammatory challenges IL-22的双重作用：在妊娠炎症期间平衡保护和风险。

F&S science

Pub Date : 2025-11-01 Epub Date: 2025-09-09 DOI: 10.1016/j.xfss.2025.09.001

Umida Ganieva Ph.D. , Mahmood Bilal Ph.D. , Ana Adam , Cecilia Pena-Rasgado Ph.D. , Wren Michaels Ph.D. , Hector Rasgado-Flores Ph.D. , Amy Thees Ph.D. , Thanh Luu M.D. , Joanne Kwak-Kim M.D. , Svetlana Dambaeva M.D., Ph.D.

Objective

To investigate the potential of recombinant interleukin-22 (rIL-22) in preventing inflammation-triggered abortion.

Design

Experimental animal study using wild type (WT, C57BL/6J) and IL-22 knockout (IL-22⁻/⁻) mice aged 7–14 weeks. Abortion was induced via intraperitoneal injection of lipopolysaccharide (LPS) on gestational day 8.5.

Subjects

Pregnant WT and IL-22⁻/⁻ mice (n = 6/group for tissue collection at gestational day 8.5+48 hours; n = 3/group for monitoring to delivery at gestational day 20.5).

Intervention(s)

Tail vein injection of recombinant IL-22 at varying doses (5, 10, or 20 µg/mouse) prior to LPS administration.

Main Outcome Measure(s)

Pregnancy outcomes (abortion vs. live delivery), endometrial mucosal architecture, epithelial barrier integrity (transepithelial electric resistance), gene expression profiles (qRT-PCR), protein distribution (IHC/IF), and immune marker levels in amniotic fluid (multiplex immunoassay).

Results

Administration of rIL-22 to IL-22⁻/⁻ mice restored endometrial mucosal architecture to levels similar to WT animals and prevented abortion in a dose-dependent manner. rIL-22 treatment rescued pregnancy in IL-22⁻/⁻ mice at 5–20 µg/mouse, whereas only ≤10 µg/mouse was effective in WT mice. High-dose rIL-22 (20 µg/mouse), when combined with LPS, negatively affected pregnancy outcomes in WT mice, and 20 µg alone resulted in adverse outcomes in both genotypes.

Conclusion

Interleukin-22 emerges as a promising therapeutic target in reproductive medicine, offering potential avenues for preserving pregnancy in the face of inflammatory challenges. However, increased levels of IL-22 could be associated with reproductive failures, emphasizing the delicate balance required for immune regulation. Our study underscores the importance of immune regulation in reproductive health and highlights IL-22 as a novel candidate for improving pregnancy outcomes in inflammatory conditions.

目的：探讨重组IL-22 （IL-22）在预防炎症性流产中的作用。设计：采用野生型（WT, C57BL/6J）和IL-22敲除型（IL-22-/-）小鼠，7-14周龄，在妊娠日（GD） 8.5时通过腹腔注射脂多糖（LPS）诱导流产，模拟妊娠中期的炎症挑战。在LPS给药之前，小鼠通过尾静脉注射不同剂量的rIL-22。lps注射后48小时采集子宫组织（n=6/组），或监测小鼠直至分娩（GD 20.5）（n=3/组）。经上皮电阻评估子宫上皮细胞培养紧密连接的运输完整性。采用qRT-PCR定量检测基因表达水平，免疫组织化学/免疫荧光检测相关抗原的组织分布。采用多重免疫分析法测定羊水中的免疫标记物。结果：给IL-22-/-小鼠注射IL-22不仅使子宫内膜粘膜结构恢复到与WT动物相似的水平，而且以剂量依赖的方式防止流产。虽然IL-22在5、10和20 μg/只小鼠剂量下可挽救IL-22-/-小鼠的妊娠，但只有较低剂量（≤10 μg/只小鼠）对WT小鼠有效。高剂量（20 μg/只）rIL-22后再注射LPS对WT小鼠妊娠结局有负面影响，单独给药20 μg rIL-22对两种基因型小鼠均有不良妊娠结局。结论：IL-22在生殖医学中是一个很有前景的治疗靶点，为在面临炎症挑战时保留妊娠提供了潜在的途径。然而，IL-22水平的升高可能与生殖失败有关，强调了免疫调节所需的微妙平衡。我们的研究强调了免疫调节在生殖健康中的重要性，并强调了il -22作为改善炎症条件下妊娠结局的新候选者。

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引用次数: 0

From the Editor-in-Chief 科学的工作就是用事实代替现象，用论证代替印象。

F&S science

Pub Date : 2025-11-01 Epub Date: 2025-10-10 DOI: 10.1016/j.xfss.2025.10.002

William H. Catherino M.D., Ph.D.

引用次数: 0

Progesterone receptor isoform modulation via enhancer activation regulates progesterone signaling in endometrial stromal cells 通过增强子激活的PGR异构体调节子宫内膜间质细胞中的孕酮信号。

F&S science

Pub Date : 2025-11-01 Epub Date: 2025-08-21 DOI: 10.1016/j.xfss.2025.08.004

Skylar G. Montague Redecke B.Sc. , Austin Bell-Hensley Ph.D. , MyeongJin Yi Ph.D. , Shuyun Li Ph.D. , Francesco J. DeMayo Ph.D.

Objective

To investigate enhancer-mediated regulation of progesterone receptor (PGR) isoforms, PGR-A and PGR-B, in human endometrial stromal cells, and to determine how isoform modulation shapes the progesterone-responsive transcriptome and cistrome relevant to endometrial function.

Design

A clustered regularly interspaced short palindromic repeats–based functional genomic screen was used to identify distal enhancers in telomerase-immortalized human endometrial stromal cells. Subsequent clustered regularly interspaced short palindromic repeats targeting of identified enhancers and the PGR promoter was used to modulate PGR isoform balance and assess functional consequences.

Subjects

None.

Exposure

Engineered endometrial stromal cells were treated with medroxyprogesterone acetate or vehicle.

Main Outcome Measures

PGR isoform expression was assessed by western blot, the progesterone-responsive transcriptome was characterized by bulk ribonucleic acid sequencing, and the PGR cistrome was characterized by Cut&Run.

Results

Two distal PGR enhancers were identified in endometrial stromal cells located approximately 60 and 220 kb upstream of the PGR transcription start site. Clustered regularly interspaced short palindromic repeats–based activation of these enhancers upregulated both PGR-A and PGR-B, whereas promoter activation primarily upregulated PGR-B. Bulk ribonucleic acid sequencing revealed that shifting the PGR isoform balance altered the progesterone-regulated transcriptome: PGR-A/B-equivalent cells exhibited proinflammatory gene signatures, whereas PGR-B-dominant cells demonstrated suppression of inflammatory signaling and altered cell cycle programs. The PGR Cut&Run profiling revealed distinct genomic binding patterns associated with each isoform profile. Integration of the PGR cistrome with chromatin interaction maps suggested that these isoforms directly regulate distinct gene subsets involved in inflammation and fibrosis. Mechanistically, estrogen receptor alpha (ESR1) indirectly activated PGR-A expression, potentially through recruitment of Forkhead box protein O1 (FOXO1) at the distal enhancer, suggesting a noncanonical, enhancer-mediated mechanism of PGR regulation.

Conclusions

Distal enhancers regulate the PGR isoform balance and shape the progesterone-responsive transcriptome in human endometrial stromal cells. This enhancer-mediated mechanism expands current models of PGR regulation beyond promoter-level control and may offer potential therapeutic targets to restore normal progesterone responsiveness in conditions marked by PGR isoform imbalance.

目的：研究增强子介导的孕激素受体（PGR）异构体PGR- a和PGR- b在人子宫内膜基质细胞中的调节作用，并确定异构体调节如何影响与子宫内膜功能相关的孕激素应答转录组和胞质。设计：使用基于crispr的功能基因组筛选来鉴定端粒酶永生化的人子宫内膜基质细胞中的远端增强子。随后使用CRISPR靶向鉴定的增强子和PGR启动子来调节PGR异构体平衡并评估功能后果。主题:没有。暴露：工程子宫内膜基质细胞用醋酸甲羟孕酮（MPA）或载体处理。主要观察指标：western blot检测PGR亚型表达，bulk RNA测序检测黄体酮应答转录组，Cut&Run检测PGR胞浆。结果：在位于PGR转录起始位点上游约60和220 kb的子宫内膜基质细胞中发现了两个远端PGR增强子。基于crispr的这些增强子的激活上调了PGR-A和PGR-B，而启动子的激活主要上调了PGR-B。大量RNA测序显示，改变PGR异构体平衡改变了黄体酮调节的转录组：PGR- a / b等效细胞表现出促炎基因特征，而PGR- b优势细胞表现出炎症信号抑制和细胞周期程序改变。PGR Cut&Run分析揭示了不同的基因组结合模式与每个异构体剖面相关。PGR细胞质与染色质相互作用图谱的整合表明，这些同工型直接调节参与炎症和纤维化的不同基因亚群。从机制上讲，ESR1间接激活了PGR- a的表达，可能是通过远端增强子FOXO1的募集，提示PGR调控的非规范、增强子介导的机制。结论：在人子宫内膜基质细胞中，远端增强子调节PGR异构体平衡并塑造黄体酮应答转录组。这种增强子介导的机制扩展了目前的PGR调节模型，超出了启动子水平的控制，并可能提供潜在的治疗靶点，以恢复PGR异构体失衡的正常黄体酮反应性。

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引用次数: 0

Transcriptome landscapes induced by estetrol, estradiol, and ethinylestradiol in primary human endometriotic stromal cells 在原发性人子宫内膜异位症基质细胞中由雌二醇、雌二醇和乙炔雌二醇诱导的转录组景观。

F&S science

Pub Date : 2025-11-01 Epub Date: 2025-06-25 DOI: 10.1016/j.xfss.2025.06.002

Marie Nakajima M.D. , Gentaro Izumi M.D., Ph.D. , Kaori Koga M.D., Ph.D. , Vita Silvana M.D. , Mohammed Elsherbini M.D., Ph.D. , Atsushi Okumura M.Sc. , Michihito Wada Ph.D. , Satoru Yamanaka Ph.D. , Yasushi Hirota M.D., Ph.D. , Yutaka Osuga M.D., Ph.D.

Objective

To compare the transcriptomes induced by estrogens used in combined oral contraceptives (COCs): estetrol (E4), ethinylestradiol (EE), and estradiol (E2) in human primary endometriotic stromal cells.

Design

Endometriotic stromal cells were cultured in the presence of E4 (10⁻⁶ mol/L), E2 (10⁻⁸ mol/L) or EE (10⁻⁹ mol/L) for 24 hours. Transcriptomes induced by E4 were compared with those induced by E2 and EE.

Subjects

Endometriotic stromal cells were obtained from patients with ovarian endometriomas.

Main Outcome Measures

Transcriptomes were examined using RNA sequencing analysis, and messenger RNA levels were measured using reverse transcription-quantitative polymerase chain reaction.

Results

Estetrol treatment resulted in 114 differentially expressed genes compared with the control (|log2fold change| >0.2). The number was higher than those differentially expressed between E4 and E2 (82 genes) and between E4 and EE (75 genes). Of the differentially expressed genes between E4 and the control, 79% (90 genes) changed in the same manner as in EE and E2. In contrast, 21% (24 genes) of the genes changed in an E4-preferential manner.

Conclusion

The present study elucidated that although E4 induces the expression of specific unique genes in endometriotic stromal cells distinct from those regulated by other estrogens, the overall gene expression profile is largely analogous to that induced by other estrogens. When differences in progestin are not considered, it is suggested that the COC containing E4 exhibits effects on endometriosis comparable with those of other COCs.

目的：比较复方口服避孕药（COCs）中使用的雌激素：雌二醇（E4）、炔雌醇（EE）和雌二醇（E2）在人原发性子宫内膜异位症基质细胞（ESCs）中诱导的转录组。设计：ESCs在E4 （10-6 mol/L）、E2 （10-8 mol/L）或EE （10-9 mol/L）存在下培养24小时。将E4诱导的转录组与E2和EE诱导的转录组进行比较。对象：从卵巢子宫内膜异位瘤患者身上获得ESCs。主要结果测量：使用RNA测序分析检测转录组，使用逆转录定量聚合酶链反应（RT-qPCR）测量mRNA水平。结果：与对照组相比，E4处理导致114个基因差异表达（|log2倍变化| >0.2）。E4与E2（82个基因）、E4与EE（75个基因）的差异表达量高于E4与E2（82个基因）的差异表达量。在E4与对照组的差异表达基因中，79%（90个基因）的变化方式与EE和E2相同。相比之下，21%（24个基因）的基因以e4优先的方式发生变化。结论：本研究表明，虽然E4在ESCs中诱导了不同于其他雌激素调控的特异性独特基因的表达，但其整体基因表达谱与其他雌激素诱导的基因表达谱在很大程度上相似。如果不考虑黄体酮的差异，则表明含有E4的COC对子宫内膜异位症的影响与其他COC相当。

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引用次数: 0

Impact of a microfluidic-based selection device on sperm deoxyribonucleic acid fragmentation and intracytoplasmic sperm injection cycles 基于微流体的选择装置对精子DNA断裂和icsi周期的影响。

F&S science

Pub Date : 2025-11-01 Epub Date: 2025-05-19 DOI: 10.1016/j.xfss.2025.05.002

Maria Luisa Pardiñas M.Sc. , Rocío Rivera-Egea Ph.D. , Jose Maria de los Santos Ph.D. , Carmen Vidal Ph.D. , Juan Giles Ph.D. , David Ortega-Jaen M.Sc. , Julia Gil M.Sc. , Angel Martin Ph.D. , Thamara Viloria Ph.D. , Maria Jose de los Santos Ph.D.

Objective

To determine the impact of a microfluidic-based sperm selection device on sperm parameters and embryo variables compared with the conventional swim-up method in sibling oocytes.

Design

Prospective observational study.

Subjects

A total of 345 oocytes were recruited from 27 couples undergoing intracytoplasmic sperm injection, including both own (n = 195) and donation (n = 150) cycles. None of the patients presented severe male factor.

Exposure

Semen sample was divided into 2 groups and processed using swim-up or the microfluidic sperm selection device (MSSD) ZyMōt. Half of the oocytes were inseminated with swim-up–selected spermatozoa, and the rest were inseminated with MSSD-selected spermatozoa. Embryo development was followed by time-lapse. Sperm deoxyribonucleic acid (DNA) fragmentation was measured using the sperm chromatin dispersion test, analyzed with ImageJ. A Mann-Whitney U test was performed for statistical analysis.

Main Outcome Measures

Sperm DNA fragmentation levels, sperm parameters, fertilization rates, embryo morphokinetics, and rate of usable blastocysts.

Results

Sperm DNA fragmentation was significantly lower in the MSSD group than in the swim-up group (20.3% vs. 11%), indicating a better selection. Analyzing separately the oocytes from the patient's own cycles, MSSD showed a significantly higher rate of usable blastocysts per fertilized oocyte. However, this difference was not observed using donated oocytes or when both cycles were combined. Embryos from the swim-up group showed a significant delay in time of pronuclear appearance and morula formation compared with those from the MSSD group, being more marked in donated oocytes. No significant differences were observed in the fertilization rate and the remaining morphokinetic times.

Conclusion

This study provides valuable information on the use of MSSD for noninvasive sperm selection. When MSSD was used, we observed a reduction in sperm DNA fragmentation and an enhancement in the number of usable embryos in our own cycles. These findings could be compatible with a reduced capacity to repair sperm damage due to poorer oocyte quality caused by advanced age.

目的：研究基于微流体的精子选择装置对兄弟姐妹卵母细胞精子参数和胚胎变量的影响，并与传统的游动方法进行比较。设计：前瞻性观察研究。对象：共从27对接受卵浆内单精子注射（ICSI）的夫妇中招募345个卵母细胞，包括自己的（n=195）和捐赠的（n=150）周期。所有患者均未出现严重的男性因素。干预/暴露：精液样本分为两组，分别使用(1)游泳或(2)微流控精子选择器（MSSD） ZyMōt进行处理。一半的卵母细胞用向上游动选择的精子受精，其余的卵母细胞用向上游动选择的精子受精。胚胎发育被延时跟踪。用精子染色质分散试验测定SDF，用ImageJ分析。采用Mann-Whitney U检验进行统计分析。主要观察指标：精子DNA碎片（SDF）水平、精子参数、受精率、胚胎形态动力学和可用囊胚率。结果：与游泳组相比，MSSD组的SDF显著降低(20.3%比11%；P=0.004)，说明选择较好。单独分析患者自身周期的卵母细胞，MSSD显示每个受精卵母细胞的可用囊胚率显着提高（P=0.041）。然而，使用捐赠的卵母细胞或两个周期联合使用时，没有观察到这种差异（P < 0.05）。与MSSD组相比，游泳组胚胎的原核出现时间（tPNa）和桑葚胚形成时间（tM）明显延迟，在捐赠卵母细胞中更为明显（P0.05）。结论：本研究为MSSD在无创精子选择中的应用提供了有价值的信息。当使用MSSD时，我们观察到在我们自己的周期中SDF的减少和可用胚胎的数量的增加。这些发现可能与高龄导致的卵母细胞质量下降导致的精子修复能力下降相一致。

{"title":"Impact of a microfluidic-based selection device on sperm deoxyribonucleic acid fragmentation and intracytoplasmic sperm injection cycles","authors":"Maria Luisa Pardiñas M.Sc. , Rocío Rivera-Egea Ph.D. , Jose Maria de los Santos Ph.D. , Carmen Vidal Ph.D. , Juan Giles Ph.D. , David Ortega-Jaen M.Sc. , Julia Gil M.Sc. , Angel Martin Ph.D. , Thamara Viloria Ph.D. , Maria Jose de los Santos Ph.D.","doi":"10.1016/j.xfss.2025.05.002","DOIUrl":"10.1016/j.xfss.2025.05.002","url":null,"abstract":"<div><h3>Objective</h3><div>To determine the impact of a microfluidic-based sperm selection device on sperm parameters and embryo variables compared with the conventional swim-up method in sibling oocytes.</div></div><div><h3>Design</h3><div>Prospective observational study.</div></div><div><h3>Subjects</h3><div>A total of 345 oocytes were recruited from 27 couples undergoing intracytoplasmic sperm injection, including both own (n = 195) and donation (n = 150) cycles. None of the patients presented severe male factor.</div></div><div><h3>Exposure</h3><div>Semen sample was divided into 2 groups and processed using swim-up or the microfluidic sperm selection device (MSSD) ZyMōt. Half of the oocytes were inseminated with swim-up–selected spermatozoa, and the rest were inseminated with MSSD-selected spermatozoa. Embryo development was followed by time-lapse. Sperm deoxyribonucleic acid (DNA) fragmentation was measured using the sperm chromatin dispersion test, analyzed with ImageJ. A Mann-Whitney <em>U</em> test was performed for statistical analysis.</div></div><div><h3>Main Outcome Measures</h3><div><span>Sperm DNA fragmentation levels, sperm parameters, fertilization rates, embryo morphokinetics, and rate of usable </span>blastocysts.</div></div><div><h3>Results</h3><div><span>Sperm DNA fragmentation was significantly lower in the MSSD group than in the swim-up group (20.3% vs. 11%), indicating a better selection. Analyzing separately the oocytes from the patient's own cycles, MSSD showed a significantly higher rate of usable blastocysts per fertilized oocyte. However, this difference was not observed using donated oocytes or when both cycles were combined. Embryos from the swim-up group showed a significant delay in time of pronuclear appearance and </span>morula formation compared with those from the MSSD group, being more marked in donated oocytes. No significant differences were observed in the fertilization rate and the remaining morphokinetic times.</div></div><div><h3>Conclusion</h3><div>This study provides valuable information on the use of MSSD for noninvasive sperm selection. When MSSD was used, we observed a reduction in sperm DNA fragmentation and an enhancement in the number of usable embryos in our own cycles. These findings could be compatible with a reduced capacity to repair sperm damage due to poorer oocyte quality caused by advanced age.</div></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"6 4","pages":"Pages 377-386"},"PeriodicalIF":0.0,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144121521","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Alcohol alters blood-testis barrier function in an in vitro model 酒精在体外模型中改变血睾丸屏障功能

F&S science

Pub Date : 2025-11-01 Epub Date: 2025-07-29 DOI: 10.1016/j.xfss.2025.07.005

R. Clayton Edenfield Ph.D. , Samuel B. Potter B.S. , Krista S. Crow Ph.D. , In Ki Cho Ph.D. , Kristen F. Easley Ph.D. , Nathalia L.M. Lara Ph.D. , Elizabeth S. Waters B.S. , Jason C. Hedges M.D., Ph.D. , Jamie O. Lo M.D. , Ina Dobrinski Ph.D. , Michael Koval Ph.D. , Charles A. Easley Ph.D.

Objective

This study aims to investigate the impact of alcohol on blood-testis barrier (BTB) integrity using a novel in vitro model and to elucidate potential nonhormonal mechanisms underlying alcohologenic reversible azoospermia.

Design

Primary rhesus macaque Sertoli cells were exposed to ethanol to evaluate dose-dependent effects on BTB integrity. Barrier function was assessed through electrical resistance and permeability assays, with recovery evaluated after a 48-hour withdrawal period. Gene expression of Sertoli cells and tight junction markers and cytokine levels associated with barrier degradation were analyzed.

Subjects

Primary rhesus macaque Sertoli cells were used.

Exposure

Clinically relevant concentrations (10, 60, and 100 mM), equivalent to 1–2 drinks to requiring hospitalization, of ethanol were used.

Main Outcome Measures

The main outcome measures were BTB integrity and permeability, assessed via transepithelial electrical resistance and Dye Flux assays, recovery of barrier function after ethanol withdrawal, gene expression changes in Sertoli cells and tight junction markers, and cytokine levels associated with barrier impairment. These metrics evaluated the functional and molecular impacts of in vitro ethanol exposure on the BTB.

Results

Ethanol exposure was associated with a dose-dependent decrease in BTB integrity, with partial recovery observed at lower concentrations (10 and 60 mM) after 48 hours of withdrawal, but not at 100 mM. Additionally, ethanol increased the expression of key Sertoli cells and tight junction genes such as SOX9 and CLDN3, and elevated cytokines associated with barrier degradation at higher concentrations.

Conclusion

Clinically relevant ethanol concentrations reversibly disrupt BTB function through a nonhormonal mechanism, with partial recovery at lower concentrations. These findings provide novel insights into the role of BTB dysfunction in alcohologenic reversible azoospermia.

目的：本研究旨在通过一种新的体外模型研究酒精对血睾丸屏障完整性的影响，并阐明酒精源性可逆性无精子症的潜在非激素机制。设计：将原代恒河猴的支持细胞暴露于乙醇中，以评估其对BTB完整性的剂量依赖性影响。通过电阻和渗透性测试评估屏障功能，并在48小时停药后评估恢复情况。分析支持细胞和紧密连接标志物的基因表达以及与屏障降解相关的细胞因子水平。实验对象：采用原代恒河猴支持细胞。暴露：使用临床相关浓度（10、60和100 mM），相当于1-2杯需要住院治疗的乙醇。主要结局指标：主要结局指标是BTB的完整性和通透性，通过经上皮电阻和染料通量测定来评估，乙醇戒断后屏障功能的恢复，支持细胞和紧密连接标记物的基因表达变化，以及与屏障损伤相关的细胞因子水平。这些指标评估了体外乙醇暴露对BTB的功能和分子影响。结果：乙醇暴露与BTB完整性的剂量依赖性下降有关，在48小时后，在较低浓度（10和60 mM）下观察到部分恢复，但在100 mM下没有。此外，乙醇增加了关键的支持细胞和紧密连接基因（如SOX9和CLDN3）的表达，并在高浓度下升高了与屏障降解相关的细胞因子。结论：临床相关的乙醇浓度通过非激素机制可逆地破坏BTB功能，并在较低浓度下部分恢复。这些发现为BTB功能障碍在酒精源性可逆性无精子症中的作用提供了新的见解。

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引用次数: 0

Fertilin peptide decreases embryo resorption and improves live birth rate in mouse 受精肽降低胚胎吸收，提高小鼠活产率。

F&S science

Pub Date : 2025-11-01 Epub Date: 2025-08-08 DOI: 10.1016/j.xfss.2025.08.001

Sophie Favier Ph.D. , Audrey L’Hostis M.Sc. , Rémi Pierre Ph.D. , Isabelle Lagoutte Ph.D. , Eric Vicaut M.D., Ph.D. , Daniel Vaiman Ph.D. , Jean Philippe Wolf M.D., Ph.D

Objective

To study the effect of the mouse Fertilin peptide, on mouse in vitro and in utero embryo development.

Design

The Fertilin peptide, a cyclic tripeptide derived from a loop of the ADAM2 protein, is already known to provide healthy pups over three generations of mice. The present prospective randomized study analyzes the in vitro and in vivo mouse embryo development using ultrasound examination, and birth rates, considering embryo resorption in mice as the equivalent of miscarriage in humans.

Subjects

B6CBA F1 female mice were crossed with C57BL6/J male mice. A total of 458 day 2 embryos were retrieved and involved in the study: 150 for in vitro study, 66 for immunofluorescence, and 242 were transferred in vivo.

Exposure

Two-cell embryos were incubated in an embryoscope without or with the Fertilin peptide at 100 μM. The embryos were then either followed up in vitro until blastocyst formation, or were transferred in utero on day 3. Their development was analyzed using ultrasound examination. Birth rates were registered.

Main Outcome Measures

The main outcomes are: (1) in vitro kinetics of embryo development in the control and Fertilin group, (2) in vivo development of embryos analyzed by ultrasound analysis. The pregnant female mice were kept separately so that newborns could be counted after they gave birth (3).

Results

In mice, Fertilin 100 μM accelerated blastocyst formation in vitro, and reduced embryo resorption in vivo from 48.6% to 33.0%. Consistently, the live birth rate was increased from 42% to 63%.

Conclusion

This study demonstrates that Fertilin peptide accelerates embryo development in vitro, reduces the resorption rate in utero, and increases the live birth rate in mice.

In the mouse model, Fertilin appears to be the first molecule able to significantly reduce miscarriage in vivo and improve live birth rate. The human molecule is currently being evaluated in an ongoing prospective, randomized, multicenter clinical trial (NCT:04954274).

目的：研究小鼠受精肽对小鼠体外和宫内胚胎发育的影响。设计：受精肽是一种从ADAM2蛋白的一个环中提取的环状三肽，已知可以为三代以上的小鼠提供健康的幼崽。本前瞻性随机研究利用超声检查分析小鼠体外和体内胚胎发育和出生率，认为小鼠胚胎吸收相当于人类流产。实验对象：B6CBA F1雌性小鼠与C57BL6/J雄性小鼠杂交。共取出458个第2天胚胎参与研究：150个用于体外研究，66个用于免疫荧光研究，242个用于体内移植。干预（用于研究临床试验）或暴露（用于观察性研究）：两个细胞胚胎在100μM下不含或含Fertilin肽的胚胎镜中孵育。然后，这些胚胎要么在体外随访直到囊胚形成，要么在第3天在子宫内移植。采用超声检查分析其发展情况。登记了出生率。主要观察指标：主要观察指标有：(1)对照组和Fertilin组胚胎体外发育动力学；(2)超声分析胚胎体内发育情况。怀孕的雌性老鼠被分开饲养，以便在它们分娩后可以计数新生儿(3)。结果：在小鼠实验中，Fertilin 100μM能促进体外胚胎形成，使体内胚胎吸收率从48.6%降低到33.0% (p)结论：本研究表明，Fertilin肽能促进体外胚胎发育，降低子宫内吸收率，提高小鼠活产率。解释：在小鼠模型中，Fertilin作为第一个能够显著减少体内流产和提高活产率的分子出现。目前，该分子正在一项前瞻性、随机、多中心临床试验中进行人体测试（NCT:04954274）。

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引用次数: 0

Utilization of a three-dimensional in vitro co-culture system to characterize embryonic mechanisms associated with implantation 利用三维体外共培养系统，描述与植入相关的胚胎机制。

F&S science

Pub Date : 2025-08-01 Epub Date: 2025-03-19 DOI: 10.1016/j.xfss.2025.03.001

Keelee J. McCarty Ph.D., Blair McCallie Ph.D., William B. Schoolcraft M.D., Mandy Katz-Jaffe Ph.D.

<div><h3>Objective</h3><div>Implantation success is dependent on timely molecular signaling to establish embryonic apposition, adhesion, and invasion. In an effort to elucidate this critical period in human reproduction, the objective of this study was to use a novel, time-lapse three-dimensional (3D) in vitro co-culture system to characterize the timing of blastocyst development on the initial stages of implantation.</div></div><div><h3>Design</h3><div>Endometrial biopsies<span> were collected from fertile oocyte<span> donors to generate individual 3D wells of separated monolayers of luminal endometrial epithelial and stromal cells<span><span> for co-culture with an individual blastocyst (n = 72; </span>maternal age = 36.3 ± 5.1 years). After 72 hours of co-culture (CytoSMART Lux3 time-lapse imaging system), blastocysts were evaluated for stage of implantation and separated into two groups: No implantation (no adhesion or invasion) and implantation (complete adhesion and invasion).</span></span></span></div></div><div><h3>Subjects</h3><div>N/A.</div></div><div><h3>Exposure</h3><div>N/A.</div></div><div><h3>Main Outcome Measures</h3><div>Immunohistochemistry<span> and targeted quantitative real-time polymerase chain reaction gene expression were performed on individual blastocysts and on exosome-purified small RNAs derived from supernatant.</span></div></div><div><h3>Results</h3><div><span><span>After successful implantation into the endometrial epithelium<span><span>, correctly timed blastocysts experienced greater duration of apposition and adhesion, delayed onset of invasion, and increased number of spontaneous blastocyst collapse compared with slower developing blastocysts. Additionally, targeted </span>gene expression analysis revealed an upward trend of implantation-promoting genes GATA </span></span>binding protein 3, </span>octamer binding transcription factor 4<span><span><span><span>, and cell death regulatory gene<span> caspase </span></span>protein 3 in correctly timed blastocysts compared with slower developing blastocysts. Interestingly, as blastocysts became more attached to the epithelium, a downward trend of </span>developmental genes<span><span> caudal-related homeobox 2 and </span>bone morphogenic protein 15 was observed. A downward trend of hsa-miR-1-3p and an upward trend of hsa-miR-34b-5p was observed in the supernatant of co-cultured blastocysts that achieved successful implantation in co-culture. Top Kyoto Encyclopedia of Genes and Genomes pathways impacted by these </span></span>microRNAs<span><span> were axon guidance, ubiquitin-mediated </span>proteolysis<span><span>, and neurotrophin </span>signaling pathway.</span></span></span></div></div><div><h3>Conclusion</h3><div>Time-lapse 3D in vitro co-culture revealed that the timing of blastocyst development is critical to the initial stages of implantation. The ability of the trophectoderm to expand, orient, and initiate apposition may contribute to the higher likelihood of

目的：胚胎着床的成功依赖于及时的分子信号来建立胚胎的附着、粘附和侵袭。为了阐明人类生殖的这一关键时期，本研究的目的是利用一种新颖的延时3D体外共培养系统来表征胚胎着床初始阶段囊胚发育的时间。设计：从可育卵母细胞供者处收集子宫内膜活检，生成单独的三维孔，由分离的单层腔内内膜上皮细胞和基质细胞组成，与单个囊胚共培养(n = 72；产妇年龄= 36.3±5.1岁)。共培养72h后（CytoSMART Lux3延时成像系统），评估囊胚着床阶段，并将其分为未着床组（无粘连、无侵袭）和着床组（完全粘连、无侵袭）。受试者：N/A干预（随机对照试验）或暴露（观察性研究）：N/A主要结果测量：对单个囊胚和从上清提取的外泌体纯化的小rna进行免疫组织化学和靶向qPCR基因表达。结果：在成功植入子宫内膜上皮后，与发育较慢的囊胚相比，正确时间的囊胚经历了更长的附着和粘附时间，侵袭延迟，自发性囊胚塌陷的数量增加。此外，靶向基因表达分析显示，与发育较慢的囊胚相比，在正确的时间内，促着床基因GATA3、OCT4和细胞死亡调控基因CASP3的表达呈上升趋势。有趣的是，随着囊胚越来越靠近上皮，发育基因CDX2和BMP15呈下降趋势。共培养成功着床的囊胚上清液中hsa-miR-1-3p呈下降趋势，hsa-miR-34b-5p呈上升趋势。受这些microRNA影响的主要KEGG通路是轴突引导、泛素介导的蛋白水解和神经营养因子信号通路。结论：延时3D体外共培养显示囊胚发育时间对着床初期至关重要。滋养外胚层的扩张、定向和启动对抗的能力可能有助于通过改变基因表达和调控途径提高成功的可能性。

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引用次数: 0

Impact of CYP19A1 genetic variations on polycystic ovary syndrome: findings from a case-control study CYP19A1基因变异对多囊卵巢综合征的影响：一项病例对照研究的结果

F&S science

Pub Date : 2025-08-01 Epub Date: 2025-03-28 DOI: 10.1016/j.xfss.2025.03.005

Hiral Chaudhary Ph.D. , Jalpa Patel Ph.D. , Nayan K. Jain Ph.D. , Sonal Panchal M.D. , Purvi Nanavati M.D. , Mala Singh Ph.D. , Naresh Laddha Ph.D. , Rushikesh Joshi Ph.D.

Objective

To study the association between CYP19A1 genetic variants and the risk of developing polycystic ovary syndrome (PCOS). The study explored the relationship between the candidate gene CYP19A1 and hyperandrogenism, as well as its interplay with obesity, in PCOS patients compared with healthy controls.

Design

A case-control study with genetic association analysis by tetra-primer amplification refractory mutation system polymerase chain reaction and biochemical analysis.

Subjects

204 women (113 PCOS patients and 91 healthy controls) were included in the present study.

Exposure

CYP19A1 variants (rs700519 and rs2236722) in PCOS women.

Main Outcome Measures

Genotypic and allelic frequencies of CYP19A1 variants (rs2236722 and rs700519) and their impact on androgen metabolism and obesity markers.

Results

The genotypic and allelic frequency of rs2236722 showed statistically significant differences between PCOS cases and controls. A significant association was observed under the dominant model, with an odds ratio of 0.34 (95% confidence interval, 0.16–0.66), as well as under the heterozygous model, where the odds ratio was 2.58 (95% confidence interval, 1.34–4.97). However, rs700519 did not reveal any significant association between the groups. A noticeable statistical difference was observed in the levels of total testosterone, dehydroepiandrosterone sulfate , prolactin, luteinizing hormone/follicle-stimulating hormone , Estradiol/total testosterone ratio, body mass index, and waist-to-hip ratio between the case and control groups . However, no variations in clinical variables were observed among genotypes within the PCOS group.

Conclusion

Our study demonstrates that the CYP19A1 rs2236722 polymorphism significantly correlates with PCOS risk, although rs700519 showed no significant association. The findings suggest that altered aromatase activity linked to rs2236722 may contribute to the hyperandrogenic phenotype observed in PCOS patients. These results enhance our understanding of the genetic basis of PCOS and may have implications for personalized treatment approaches.

目的：探讨CYP19A1基因变异与PCOS发病风险的关系。该研究探讨了候选基因CYP19A1与多囊卵巢综合征患者与健康对照者之间的关系，以及它与肥胖的相互作用。设计：采用Tetra ARMS PCR和生化分析进行遗传关联分析的病例对照研究。对象：204名女性（113名多囊卵巢综合征患者和91名健康对照）纳入本研究。主要结局指标：CYP19A1变异（rs2236722和rs700519）的基因型和等位基因频率及其对雄激素代谢和肥胖标志物的影响。结果：rs2236722基因型及等位基因频率在PCOS病例与对照组间差异有统计学意义(p)结论：本研究表明CYP19A1 rs2236722多态性与PCOS风险显著相关，而rs700519多态性与PCOS风险无显著相关性。研究结果表明，与rs2236722相关的芳香酶活性的改变可能有助于PCOS患者观察到的高雄激素表型。这些结果增强了我们对多囊卵巢综合征遗传基础的理解，并可能对个性化治疗方法产生影响。

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引用次数: 0

Relationship between early and mid-lateal phase progesterone levels and the DNA methylation of WOI-related genes in cervical secretions in fresh and frozen IVF cycles 孕激素水平与新鲜和冷冻IVF周期宫颈分泌物中woi相关基因DNA甲基化的关系

F&S science

Pub Date : 2025-08-01 Epub Date: 2025-06-25 DOI: 10.1016/j.xfss.2025.06.001

Cemil Kaya M.D. , Kübra Nur Kaplan İlhan M.Sc. , Bala Gur Dedeoglu Ph.D. , Mehmet Erdem M.D. , Ahmet Erdem M.D.

Objective

To study the association between serum progesterone (P4) levels on the day of embryo transfer (ET) and the deoxyribonucleic acid methylation of window of implantation-related genes in cervical secretions, in both fresh and frozen ET cycles among women undergoing in vitro fertilization (IVF) treatment.

Design

Single-center retrospective cohort study.

Subjects

Women undergoing ET cycles.

Exposure

Cervicovaginal materials have been used as a noninvasive surrogate in investigations of endometrial conditions and progesterone levels on the day of ET.

Main Outcome Measures

Progesterone levels and methylation changes.

Results

A total of 40 eligible women undergoing IVF-ET cycles were evaluated. On the basis of methylation analyses of CXCL14, EMCN, RARRES1, PAEP, THBS2, and GPX3, no statistically significant correlation was found with the median serum P4 levels on the day of ET. When participants were grouped into quartiles on the basis of P4 levels on the day of ET, no statistically significant differences were observed in the methylation levels of CXCL14, SERPING1, EMCN, RARRES1, PAEP, THBS2, and GPX3. The median P4 levels were significantly lower in the thawed ET group (median, 21.4 ng/mL; interquartile range, 13.3–24.0) than in the fresh transfer group (median, 24.0 ng/mL; interquartile range, 18.0–45.0). No significant differences in gene methylation levels were observed between fresh and thawed transfer types, except for THBS2, which showed significantly lower methylation levels in the thawed group than in the fresh group. Although the median P4 levels were lower on day 3 transfers (16.6 ng/mL) than on day 5 transfers (22.4 ng/mL), the difference was not statistically significant. Similarly, no significant differences in methylation levels were detected when comparing day 3 vs. day 5 ET groups.

Conclusion

No association was found between early and mid-luteal phase P4 levels and methylation changes of implantation-related genes in cervical secretions during both fresh and frozen IVF cycles.

目的：研究体外受精（IVF）妇女胚胎移植当日血清孕酮水平与新鲜和冷冻胚胎移植周期宫颈分泌物中植入窗口（WOI）相关基因DNA甲基化的关系。设计：单中心回顾性队列研究对象：接受胚胎移植周期的妇女。暴露：宫颈阴道材料已被用作子宫内膜状况和孕激素水平的非侵入性替代物。主要结局指标：黄体酮水平、甲基化变化。结果：共评估了40名接受体外受精（IVF）胚胎移植周期的符合条件的妇女。通过对CXCL14、EMCN、RARRES1、PAEP、THBS2、GPX3的甲基化分析，发现胚胎移植当天血清中位孕酮（P4）水平与CXCL14、EMCN、RARRES1、PAEP、THBS2、GPX3的甲基化差异无统计学意义（p < 0.05）。当参与者根据胚胎移植当天的P4水平分为四分位数时，CXCL14、SERPING1、EMCN、RARRES1、PAEP、THBS2和GPX3的甲基化水平无统计学差异（所有比较p < 0.05）。解冻胚胎移植组P4水平中位数显著降低(中位数：21.4 ng/mL；四分位数间距[IQR]: 13.3-24.0)与新鲜转移组相比(中位数：24.0 ng/mL；IQR: 18.0-45.0) （p = 0.039）。除THBS2基因甲基化水平在解冻组显著低于新鲜组（p = 0.013）外，新鲜和解冻转移类型之间的基因甲基化水平无显著差异。虽然第3天的P4中位数水平（16.6 ng/mL）低于第5天的22.4 ng/mL，但差异无统计学意义（p < 0.05）。同样，在比较第3天和第5天胚胎移植组时，甲基化水平没有显著差异。结论：在新鲜和冷冻试管婴儿周期中，黄体前期和中期孕酮水平与宫颈分泌物中植入相关基因的甲基化变化没有关联。

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引用次数: 0