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Enzymatic and Genetic Evaluation of Butyrylcholinesterase in Male Infertility. 男性不育症中丁基胆碱酯酶的酶学和遗传学评价。
Pub Date : 2026-01-07 DOI: 10.1016/j.xfss.2025.12.008
Rabia Habib, Khulah Sadia, Syed Nurulain, Zuha Sarwar, Aleena Rafique, Mahnoor Atequite, Sabir Hussain, Maria Arshad, Ammara Younas, Sajid Mehmood, Syed Shah

Objective: To investigate the potential role of butyrylcholinesterase enzyme activity and tentative association of BChE gene SNPs polymorphisms (rs3495, rs1803274) with risk of male infertility. Male infertility is a highly prevalent multifactorial condition with complex heterogeneous phenotypic spectrum. A significant global variation has been observed in the prevalence of male infertility contributing to approximately 20-70% of overall infertility. An interplay of genetic, hormonal, environmental and lifestyle factors are involved in the development of male infertility. Butyrylcholinesterase (BChE), an enzyme involved in oxidative stress and sperm function may play a role in infertility, but its genetic and enzymatic profiles in male infertility are unexplored.

Study design: A case control/observational study was conducted enrolling fifty-five fertile and infertile male individuals.

Subjects: Sixty-five fertile and infertile males volunteers were included in the study EXPOSURE: Volunteers were clinically diagnosed for infertility.

Main outcome measures: Plasma BChE activity was estimated spectrophotometrically by Ellman's method. SNP genotyping of BChE gene variants was performed by PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) and Tetra ARMS-PCR (tetra-primer Amplification Refractory Mutation System- PCR).

Results: Infertile group exhibited significantly lower BChE activity as compared to the fertile group (p≤0.01). In addition, notable genetic association of BChE variants with risk of male infertility was identified. BChE rs3495 showed significant correlation in dominant model (OR = 21.67, p = 0.0001) and allelic distribution (OR = 3.30, p = 0.0002) while rs1803274 showed robust association in all models (p = <0.0001).

Conclusion: This is the first study linking BChE variants and reduced BChE enzymatic activity with male infertility. These findings suggest BChE as a potential biomarker and therapeutic target for idiopathic infertility.

目的:探讨丁基胆碱酯酶活性和BChE基因snp多态性(rs3495、rs1803274)与男性不育风险的关系。男性不育症是一种非常普遍的多因素疾病,具有复杂的异质性表型谱。男性不育症的患病率在全球范围内存在显著差异,约占总体不育症的20-70%。遗传、激素、环境和生活方式等因素的相互作用与男性不育的发展有关。丁基胆碱酯酶(BChE)是一种参与氧化应激和精子功能的酶,可能在不育中发挥作用,但其在男性不育中的遗传和酶谱尚不清楚。研究设计:进行病例对照/观察性研究,纳入55名可生育和不育男性个体。研究对象:65名有生育能力和不育的男性志愿者被纳入研究。暴露:志愿者被临床诊断为不育。主要观察指标:血浆BChE活性用Ellman法分光光度法测定。采用PCR- rflp(聚合酶链反应-限制性片段长度多态性)和Tetra ARMS-PCR(四引物扩增难突变系统-PCR)对BChE基因变异进行SNP基因分型。结果:不育组BChE活性明显低于可育组(p≤0.01)。此外,BChE变异与男性不育风险的显著遗传关联被确定。BChE rs3495在显性模型(OR = 21.67, p = 0.0001)和等位基因分布(OR = 3.30, p = 0.0002)中表现出显著相关性,而rs1803274在所有模型中表现出显著相关性(p =结论:这是首次将BChE变异和BChE酶活性降低与男性不育联系起来的研究。这些发现表明BChE是特发性不孕症的潜在生物标志物和治疗靶点。
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引用次数: 0
Functional stability of the second mitotic spindle in blastomeres and its association with multinucleation repair 卵裂球第二纺锤体的功能稳定性及其与多核修复的关系。
Pub Date : 2026-01-01 DOI: 10.1016/j.xfss.2025.10.003
Taichi Sakaguchi M.D. , Hiromitsu Shirasawa M.D., Ph.D. , Yuki Ono M.D., Ph.D. , Kazumasa Takahashi M.D., Ph.D. , Mayumi Goto A.S. , Motonari Okabe M.D., Ph.D. , Chika Ariake M.D. , Akari Tsuya M.D. , Takeo Hirakawa M.D., Ph.D. , Takuya Iwasawa M.D., Ph.D. , Ayaka Fujishima M.D., Ph.D. , Yohei Onodera M.D., Ph.D. , Tae Sugawara M.D., Ph.D. , Kenichi Makino M.D., Ph.D. , Hiroshi Miura M.D., Ph.D. , Noritaka Fukunaga M.D., Ph.D. , Yoshimasa Asada M.D., Ph.D. , Yukiyo Kumazawa M.D., Ph.D. , Yukihiro Terada M.D., Ph.D.

Objective

To study mitotic spindle morphology and chromosomal segregation in second mitosis.

Design

Live-cell imaging of deoxyribonucleic acid (DNA) and tubulin during the second mitosis of 21 freeze-thawed human two-pronuclear stage embryos was performed to analyze spindle morphology and chromosome segregation dynamics. Furthermore, chromosomal aneuploidy was assessed in all cells of the embryos that developed to the blastocyst stage.

Subjects

Twenty-one freeze-thawed human two-pronuclear embryos.

Exposure

We analyzed live-imaging videos of DNA and microtubules during the second division of 21 human 2-cell embryonic blastomeres. We further analyzed the association between the results of preimplantation genetic testing for aneuploidy of all cells in the observed embryos after development into blastocysts and the nuclear status at the early embryonic stage.

Main Outcome Measures

Spindle morphology during the second mitosis, chromosomal segregation patterns, nuclear status of daughter blastomeres, and chromosomal aneuploidy of all cells in blastocysts derived from the observed embryos.

Results

Multinucleation rate in daughter nuclei was significantly lower after the second than after the first mitosis (22% vs. 56%). Neither defocusing of spindle poles—which was prominent in the first mitosis—nor chromosomal segregation abnormalities, such as misalignment of metaphase chromosomes and lagging chromosomes during chromosomal segregation, were observed. There was no association between the nuclear status at the 2- and 4-cell stages of the observed embryos that had progressed to the blastocyst stage and the preimplantation genetic testing for aneuploidy results of all cells in the blastocysts.

Conclusion

Observations of mitotic spindle morphology and chromosomal segregation behavior revealed that second mitosis is less prone to chromosomal segregation errors than first mitosis. This suggests that segregation errors occurring during the first mitosis may be corrected during the second mitosis. Furthermore, there was no association between the nuclear status during early embryonic divisions and the chromosomal status of blastocyst cells, suggesting the presence of a mechanism that corrects chromosomal abnormalities during early development.
目的:研究二次有丝分裂纺锤体形态和染色体分离。设计:对21个冷冻解冻的人类双核(2PN)期胚胎进行第二次有丝分裂时的DNA和微管蛋白活细胞成像,分析纺锤体形态和染色体分离动力学。此外,在发育到囊胚期的所有胚胎细胞中,染色体非整倍性被评估。研究对象:21个冷冻解冻的人类2PN胚胎。暴露:我们分析了21个人类2细胞胚胎卵裂球第二次分裂期间DNA和微管的实时成像视频。我们进一步分析了胚胎发育成囊胚后所有细胞的着床前非整倍体基因检测结果与胚胎早期核状态的相关性。主要观察指标:第二次有丝分裂时的纺锤体形态,染色体分离模式,子卵裂球的核状态,以及从观察到的胚胎中获得的囊胚中所有细胞的染色体非整倍体。结果:第二次有丝分裂后的子核多核率明显低于第一次有丝分裂后的子核多核率(22% vs. 56%: p = 0.03)。在第一次有丝分裂中没有纺锤体极离焦,也没有观察到染色体分离异常,如染色体分离过程中中期染色体和滞后染色体的错位。观察到的已进入囊胚期的2细胞期和4细胞期胚胎的核状态与囊胚中所有细胞的PGT-A结果之间没有关联。结论:对有丝分裂纺锤体形态和染色体分离行为的观察表明,第二次有丝分裂比第一次有丝分裂更不容易发生染色体分离错误。这表明在第一次有丝分裂期间发生的分离错误可能在第二次有丝分裂期间得到纠正。此外,胚胎早期分裂期间的核状态与囊胚细胞的染色体状态之间没有关联,这表明存在一种纠正早期发育期间染色体异常的机制。
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引用次数: 0
Environmental factors accelerate epigenetic aging of sperm via mechanistic target of rapamycin/blood-testis barrier mechanism 环境因素通过雷帕霉素/血睾丸屏障机制加速精子表观遗传老化。
Pub Date : 2026-01-01 DOI: 10.1016/j.xfss.2025.10.001
Olatunbosun Arowolo Ph.D. , Oladele A. Oluwayiose Ph.D. , Jiahui Zhu M.D. , Oleg Sergeyev M.D., Ph.D. , Emily Houle M.S. , J Richard Pilsner Ph.D. , Alexander Suvorov Ph.D., Sc.D.

Objective

To identify a common molecular mechanism of sperm epigenome reprograming by environmental factors by testing whether mechanistic target of rapamycin (mTOR)–dependent and mTOR-independent disruption of the blood-testis barrier (BTB) by environmental factors (heat stress [HS] and cadmium [Cd] exposure, respectively) accelerate epigenetic aging of sperm in a mouse model.

Design

Using Infinium methylation array, we developed a murine sperm epigenetic clock model and used it to assess epigenetic age shifts in adult mice exposed to environmental factors for the duration of two spermatogenesis cycles.

Subjects

C57BL/6 mice.

Exposure

Short-term acute intermittent whole-body HS protocol was designed to mimic human HS during heat waves. The Cd exposure group was treated with 1 μL/g body weight of water solution of CdCl2.

Main Outcome Measures

The activation of mTOR complexes by both stressors was analyzed using enzyme-linked immunosorbent assay. Effects of HS, Cd, and aging on sperm deoxyribonucleic acid methylation were compared using bioinformatic tools.

Results

We demonstrate that both mTOR-dependent BTB disruption by HS and mTOR-independent BTB disruption by Cd exposure accelerate sperm epigenetic aging, resulting in similar changes to sperm deoxyribonucleic acid methylation patterns, including changes in methylation of genes involved in embryonic development and neurodevelopment.

Conclusion

The mTOR/BTB mechanism is a novel pathway through which environmental and other stressors influence sperm epigenetic aging. This pathway is a potential therapeutic target for the mitigation of environmental effects on epigenetic programs in sperm.
背景:精子表观遗传学研究表明,精子DNA甲基化模式可能受到一系列环境因素、健康状况和衰老的影响。研究还表明,这些变化转化为不良的妊娠结局和后代的不利健康。既往研究表明,rapamycin (mTOR)/血睾丸屏障(BTB)机制的机制靶点参与了精子表观遗传衰老速率的调控,其中mTOR复合物1活性的增加打开了BTB并加速了表观遗传衰老,而mTOR复合物2活性的增加则产生了相反的结果。目的:通过检测环境因素(分别为热应激[HS]和镉[Cd]暴露)对BTB的mtor依赖型和mtor非依赖型破坏是否会加速小鼠精子表观遗传衰老,从而确定受环境因素影响的精子表观基因组重编程的共同分子机制。设计:利用Infinium甲基化阵列,我们建立了一个小鼠精子表观遗传时钟模型,并利用它来评估暴露于环境因素的成年小鼠在两个精子发生周期内的表观遗传年龄变化。实验对象:C57BL/6小鼠。暴露:短期急性间歇性全身HS方案被设计来模拟热浪期间的人类HS。Cd暴露组以1 ul/g体重的CdCl2水溶液处理。主要观察指标:采用ELISA法分析两种应激源对mTOR复合物的激活作用。利用生物信息学工具比较HS、Cd和衰老对精子DNA甲基化的影响。研究结果表明,HS对mtor依赖性BTB的破坏和Cd暴露对mtor非依赖性BTB的破坏都加速了精子的表观遗传老化,导致精子DNA甲基化模式发生类似的变化,包括与胚胎发育和神经发育有关的基因甲基化的变化。结论:mTOR/BTB机制是环境和其他应激因素影响精子表观遗传老化的新途径。该途径是减轻环境对精子表观遗传程序影响的潜在治疗靶点。
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引用次数: 0
Lactobacillus reuteri protects against heat stress-induced testicular dysfunction by modulating oxidative stress and autophagy pathways in rats 罗伊氏乳杆菌通过调节氧化应激和自噬途径防止大鼠热应激诱导的睾丸功能障碍
Pub Date : 2026-01-01 DOI: 10.1016/j.xfss.2025.10.004
Samira Parsaeinejad M.Sc. , Vahid Nejati Ph.D. , Mazdak Razi Ph.D. , Amir Tokmechi Ph.D.

Objective

To investigate the potential of Lactobacillus reuteri (Lr) in ameliorating heat stress (HS)–induced testicular damage through modulation of oxidative stress and autophagy.

Design

Experimental controlled study involving adult male Wistar rats exposed to HS and/or probiotic treatment.

Subjects

Adult male Wistar rats divided into six groups: control, Lr-only (LrP-sole), HS-only (37 °C and 40 °C), and Lr-treated HS groups.

Intervention

Heat stress was induced by immersing the scrotal area in a water bath at 37 °C or 40 °C for 20 minutes daily over 42 days. Lactobacillus reuteri probiotic (5 × 108 CFU/mL) was administered orally throughout the experimental period.

Main Outcome Measures

Sperm count, motility, viability, nuclear maturity, and deoxyribonucleic acid integrity; testicular histopathology; total antioxidant capacity and total oxidant status; expression of autophagy-related genes (p62, Atg7, Beclin-1, and LC3-I) by quantitative reverse transcriptase polymerase chain reaction; and LC3-I/II protein levels by immunohistochemistry.

Results

Heat stress significantly impaired spermatogenesis, spermiogenesis, and sperm quality, with elevated oxidative and autophagic stress, particularly at 40 °C. L. reuteri probiotic treatment significantly improved sperm parameters, restored spermatogenic activity, reduced total oxidant status, and enhanced antioxidant capacity. Moreover, Lr modulated the expression of p62, Atg7, Beclin-1, and LC3-I/II, indicating regulation of autophagic pathways and mitigation of oxidative stress within the testicular microenvironment.

Conclusion

Lactobacillus reuteri confers robust protection against HS-induced testicular dysfunction, likely through modulation of oxidative stress and autophagy. These findings highlight the therapeutic potential of probiotics as a noninvasive intervention to counteract heat-induced male infertility.
目的探讨罗伊氏乳杆菌(Lactobacillus reuteri, Lr)通过调节氧化应激和自噬来改善热应激(HS)诱导的睾丸损伤的潜力。实验对照研究涉及暴露于HS和/或益生菌治疗的成年雄性Wistar大鼠。实验对象成年雄性Wistar大鼠分为6组:对照组、纯lr组(单lr组)、纯HS组(37°C和40°C)和低lr处理HS组。干预:将阴囊区域浸泡在37°C或40°C的水浴中,每天20分钟,持续42天,诱导热应激。整个试验期间口服罗伊氏乳杆菌益生菌(5 × 108 CFU/mL)。主要观察指标:精子数量、活力、活力、核成熟度和脱氧核糖核酸完整性;睾丸组织病理学;总抗氧化能力和总氧化状态;通过定量逆转录酶聚合酶链反应表达自噬相关基因(p62、Atg7、Beclin-1和lc3 -1);和LC3-I/II蛋白水平。结果低温胁迫显著损害精子发生、精子发生和精子质量,氧化和自噬应激升高,特别是在40°C时。罗伊氏乳杆菌益生菌处理显著改善精子参数,恢复生精活性,降低总氧化状态,增强抗氧化能力。此外,Lr还可调节p62、Atg7、Beclin-1和LC3-I/II的表达,表明其可调节睾丸微环境中的自噬途径并减轻氧化应激。结论罗伊氏乳杆菌对hs诱导的睾丸功能障碍具有强大的保护作用,可能通过调节氧化应激和自噬来实现。这些发现强调了益生菌作为一种非侵入性干预来对抗热诱导的男性不育的治疗潜力。
{"title":"Lactobacillus reuteri protects against heat stress-induced testicular dysfunction by modulating oxidative stress and autophagy pathways in rats","authors":"Samira Parsaeinejad M.Sc. ,&nbsp;Vahid Nejati Ph.D. ,&nbsp;Mazdak Razi Ph.D. ,&nbsp;Amir Tokmechi Ph.D.","doi":"10.1016/j.xfss.2025.10.004","DOIUrl":"10.1016/j.xfss.2025.10.004","url":null,"abstract":"<div><h3>Objective</h3><div>To investigate the potential of <em>Lactobacillus reuteri</em> (Lr) in ameliorating heat stress (HS)–induced testicular damage through modulation of oxidative stress and autophagy.</div></div><div><h3>Design</h3><div>Experimental controlled study involving adult male Wistar rats exposed to HS and/or probiotic treatment.</div></div><div><h3>Subjects</h3><div>Adult male Wistar rats divided into six groups: control, Lr-only (LrP-sole), HS-only (37 °C and 40 °C), and Lr-treated HS groups.</div></div><div><h3>Intervention</h3><div>Heat stress was induced by immersing the scrotal area in a water bath at 37 °C or 40 °C for 20 minutes daily over 42 days. <em>Lactobacillus reuteri</em> probiotic (5 × 10<sup>8</sup> CFU/mL) was administered orally throughout the experimental period.</div></div><div><h3>Main Outcome Measures</h3><div>Sperm count, motility, viability, nuclear maturity, and deoxyribonucleic acid integrity; testicular histopathology; total antioxidant capacity and total oxidant status; expression of autophagy-related genes (p62, Atg7, Beclin-1, and LC3-I) by quantitative reverse transcriptase polymerase chain reaction; and LC3-I/II protein levels by immunohistochemistry.</div></div><div><h3>Results</h3><div>Heat stress significantly impaired spermatogenesis, spermiogenesis, and sperm quality, with elevated oxidative and autophagic stress, particularly at 40 °C. <em>L. reuteri</em> probiotic treatment significantly improved sperm parameters, restored spermatogenic activity, reduced total oxidant status, and enhanced antioxidant capacity. Moreover, Lr modulated the expression of p62, Atg7, Beclin-1, and LC3-I/II, indicating regulation of autophagic pathways and mitigation of oxidative stress within the testicular microenvironment.</div></div><div><h3>Conclusion</h3><div><em>Lactobacillus reuteri</em> confers robust protection against HS-induced testicular dysfunction, likely through modulation of oxidative stress and autophagy. These findings highlight the therapeutic potential of probiotics as a noninvasive intervention to counteract heat-induced male infertility.</div></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"7 1","pages":"Pages 35-48"},"PeriodicalIF":0.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145915423","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
Dose-dependent modulation of luteinizing hormone receptor expression in human granulosa cells and progesterone production during ovarian stimulation 卵巢刺激过程中人颗粒细胞LH受体表达和黄体酮产生的剂量依赖性调节。
Pub Date : 2026-01-01 DOI: 10.1016/j.xfss.2025.11.002
Barbara Lawrenz Ph.D. , Sahar Ghafari D.V.M. , Erkan Kalafat Ph.D. , Araz Raberi Ph.D. , Jonalyn Edades M.Sc. , Bhanu Kalra Ph.D. , Noreen Hourani M.Sc. , Virginia Ferracuti M.Sc. , Asina Bayram M.Sc. , Human Fatemi Ph.D.

Objective

To assess the impact of different recombinant follicle-stimulating hormone (recFSH) dosages in ovarian stimulation (OS) on luteinizing hormone (LH)–receptor and FSH-receptor expression in granulosa cells (GCs) and progesterone level.

Design

Prospective interventional study (07/2023–02/2024).

Subjects

Six volunteers with regular cycles and normal ovarian reserve undergoing two OS cycles for oocyte vitrification as a means of elective fertility preservation.

Exposure

Different recFSH dosages (150 IU vs. 300 IU) in two OS cycles, performed in a gonadotropin-releasing hormone–antagonist protocol.

Main Outcome Measures

The LH receptor expression on GCs.

Results

Volunteers had a median age of 34 years (range: 28–36), a median antimüllerian hormone–level of 2.28 ng/mL (interquartile range [IQR]: 2.41–3.35) and a median antral follicle count of 13.5 (IQR:12–14) in the 150 IU and 13 (12–15) in the 300 IU OS cycle. Median total recFSH dosages were 1650 IU (IQR: 1350–1800IU)/3150 IU (IQR: 3000–3300) in the 150 IU/300 IU OS cycle, respectively. The number of retrieved cumulus-oocyte-complexes was (median and IQR) 11.5 (10–14)/12.5 (7–15), the number of mature oocytes (median and IQR) of 10.5 (7–12)/10 (6–12), respectively, without a statistically significant difference.
Besides the stimulation start, serum FSH levels of the 300 IU-cycle were significantly higher compared with the 150 IU-cycle. Estradiol levels were significantly higher in the 300 IU-group on day 5 and at follicle size of 14–16 mm. Progesterone (P4) levels were not statistically significantly different between the stimulation cycles.
The LH receptor expression in the GCs at oocyte-pick up was significantly higher and FSH-receptor expression was significantly lower in the 300 IU-group compared with the 150 IU-group. The P4 levels on oocyte-pick up day were significantly associated with LH receptor levels after adjusting for the recFSH dose. Furthermore, a significant and independent association of the recFSH dose between the P4 levels and the LH receptor levels was seen on the day of oocyte retrieval.

Conclusion

Higher recFSH dosages during OS are associated with increased LH receptor expression, and LH receptor count had a significant association with serum P4 levels.
目的:探讨卵巢刺激(OS)中不同重组FSH (rec)剂量对颗粒细胞(GC) lh受体和FSH受体表达及孕酮水平的影响。设计:前瞻性介入研究(2007 /2023- 2024 / 02)。受试者:6名月经周期正常、卵巢储备正常的志愿者,接受2个周期的卵细胞玻璃化冷冻,作为选择性保留生育能力的手段。暴露:在gnrh拮抗剂方案中,在两个OS周期中不同的recFSH剂量(150IU vs 300IU)。主要观察指标:GC中lh受体的表达。结果:志愿者的中位年龄为34岁(范围:28-36岁),抗穆勒氏激素水平中位数为2.28ng/ml (IQR: 2.41-3.35),窦泡计数中位数为13.5 (IQR:12-14), 300iu - os周期中位数为13(12-15)。在150IU / 300IU的os周期中,中位总recfsh剂量分别为1650IU (IQR: 1350-1800IU) / 3150IU (IQR: 3000-3300IU)。检索到的卵丘-卵母细胞复合体数量(中位数(IQR))分别为11.5(10-14)/ 12.5(7-15),成熟卵母细胞数量(中位数和IQR)分别为10.5(7-12)/ 10(6-12),差异无统计学意义。除了刺激开始外,300iu周期的血清fsh水平显著高于150iu周期(p结论:OS期间较高的refsh剂量与LH受体表达增加有关,LH受体计数与血清p4水平显著相关。
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引用次数: 0
Complementary therapeutic actions of Shikonin and Indole propionic acid ameliorate diabetic infertility via distinct antioxidant and metabolic pathways 紫草素和吲哚丙酸通过不同的抗氧化和代谢途径改善糖尿病不孕症的互补治疗作用。
Pub Date : 2026-01-01 DOI: 10.1016/j.xfss.2025.11.001
Imran Tarique Ph.D., Ali Haider M.S., Fatima Nawazish M.S.

Objective

To evaluate the effect of Shikonin, a potent direct reactive oxygen species (ROS) scavenger, and Indole propionic acid (IPA), a microbiome-derived metabolite, on epididymis and sperm morphology and function in diabetic rats.

Design

A rat model of Type 2 diabetes mellitus (T2DM) was induced in male Wistar rats using a high-fat diet and streptozotocin (50 mg/kg). The animals were subsequently divided into seven groups: Control, T2DM, T2DM + IPA (50 mg/kg), T2DM + Shikonin (0.5 mg/kg), T2DM + Metformin (50 mg/kg), IPA-only, and Shikonin-only. Treatments were administered orally for 4 weeks. We assessed histological integrity, sperm motility, and viability (via computer-assisted sperm analysis), serum lipid profiles, epididymal antioxidant activity (superoxide dismutase, malondialdehyde, and catalase), and apoptotic marker (cytochrome C, caspase-9, and caspase-3) and metabolic signature (fibroblast growth factor 21 [FGF21]) expression via quantitative reverse transcription polymerase chain reaction.

Subjects

Male Wistar albino rats (n = 42, Age: 8–12 weeks old, 200 ± 2.4 g).

Intervention

Streptozotocin, Shikonin, Indole propionic acid.

Main Outcome Measures

Type 2 diabetes mellitus profoundly impairs male fertility, with oxidative stress in the epididymis being a key driver of sperm damage.

Results

Streptozotocin-induced diabetic rats exhibited increased oxidative damage, characterized by an impaired antioxidant system, high expression of apoptotic markers, and reduced metabolic gene expression. These effects ultimately damaged epididymis tissue and resulted in poor sperm quality. Shikonin and IPA treatments significantly attenuated oxidative stress and apoptosis, restored antioxidant activity and lipid profiles, and improved sperm parameters. Both treatments enhanced metabolic balance, as indicated by improved FGF21 expression, and restored epididymal histoarchitecture. Shikonin was superior in improving the epididymal somatic index and sperm motility, whereas IPA was more effective in normalizing FGF21 expression and lipid profiles, highlighting its primary metabolic role.

Conclusion

Our findings reveal that Shikonin and IPA protect against diabetic reproductive damage through complementary pathways. This work not only introduces promising natural therapeutic strategies but also provides a mechanistic framework for their combined or targeted use in managing diabetic infertility.
目的:探讨紫草素和微生物代谢物吲哚丙酸对糖尿病大鼠附睾和精子形态及功能的影响。设计:采用高脂饮食和链脲佐菌素(50 mg/kg)诱导雄性Wistar大鼠2型糖尿病模型。随后将动物分为7组:对照组、T2DM组、T2DM + IPA组(50 mg/kg)、T2DM +紫草素组(0.5 mg/kg)、T2DM +二甲双胍组(50 mg/kg)、单用IPA组和单用紫草素组。口服治疗4周。我们通过qRT-PCR评估了组织完整性、精子活力和活力(通过CASA)、血脂谱、附睾抗氧化活性(SOD、MDA、过氧化氢酶)、凋亡标志物(细胞色素C、caspase-9、caspase-3)和代谢特征(FGF21)的表达。动物(S):雄性Wistar白化大鼠(n = 42,年龄:8-12周龄,200±2.4 g)干预(S):链脲佐菌素,紫草素,吲哚丙酸主要结果测量(S): T2DM严重损害男性生育能力,附睾氧化应激是精子损伤的关键驱动因素。结果:链脲佐菌素诱导的糖尿病大鼠表现出增加的氧化损伤,其特征是抗氧化系统受损,凋亡标记物高表达,代谢基因表达降低。这些影响最终损害附睾组织,导致精子质量差。紫草素和吲哚丙酸处理显著减轻了氧化应激和细胞凋亡,恢复了抗氧化活性和脂质谱,改善了精子参数。通过改善FGF-21的表达,这两种治疗都增强了代谢平衡,并恢复了附睾组织结构。紫草素在改善附睾体指数和精子活力方面具有优势,而IPA在调节FGF21表达和脂质谱方面更为有效,突出了其主要的代谢作用。结论:紫草素和IPA通过互补途径对糖尿病生殖损伤具有保护作用。这项工作不仅介绍了有前途的自然治疗策略,而且为其联合或靶向治疗糖尿病性不孕症提供了机制框架。
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引用次数: 0
Deciphering estrogen receptor alpha–driven transcription in human endometrial stromal cells via transcriptome, cistrome, and integration with chromatin landscape 通过转录组、池质和染色质景观整合,解读人类子宫内膜基质细胞中esr1驱动的转录。
Pub Date : 2026-01-01 DOI: 10.1016/j.xfss.2025.08.002
Skylar G. Montague Redecke B.Sc. , Austin Bell-Hensley Ph.D. , Shuyun Li Ph.D. , MyeongJin Yi Ph.D. , Akshadha Jain , Abdull J. Massri Ph.D. , Francesco J. DeMayo Ph.D.

Objective

To investigate estrogen receptor gene 1 (ESR1) and estrogen-driven transcription in human endometrial stromal cells.

Design

RNA sequencing (RNA-seq) and Cleavage Under Targets and Release Using Nuclease (Cut&Run) were performed on telomerase-immortalized human endometrial stromal cells with Clustered Regularly Interspaced Short Palindromic Repeats–mediated ESR1 activation. Hi-C-based chromatin architecture analysis (H3K27ac HiChIP) was conducted in primary endometrial stromal cells.

Subjects

Biopsies from two healthy, reproductive-aged volunteers with regular menstrual cycles and no history of gynecological malignancies.

Exposure

The ESR1-activated and control endometrial stromal cells were treated with estradiol (E2) or vehicle. Primary endometrial stromal cells were treated with vehicle or a decidualization cocktail.

Main Outcome Measures

Differential gene expression analysis (RNA-seq) identified ligand-independent and -dependent ESR1 activity. Cut&Run profiled ESR1 genomic binding in ESR1-activated cells. H3K27ac HiChIP mapped hormone-induced changes in chromatin looping in primary cells.

Results

Among seven tested guide RNAs (gRNA), the ESR1-3 gRNA induced robust ESR1 activation and restored E2 responsiveness. Bulk RNA-seq revealed both ligand-dependent and -independent ESR1 transcriptional programs regulating inflammation, proliferation, and cancer-related pathways. Notably, 72% of differentially expressed genes overlapped with genes active in human endometrial tissue during the proliferative estrogen-dominant phase, supporting their physiological relevance. The Cut&Run-seq identified genome-wide ESR1 binding sites, with most binding sites located at distal regulatory elements. Integration of Cut&Run data with H3K27ac HiChIP chromatin loops linked distal ESR1 binding sites to gene promoters, including genes involved in decidualization (e.g., FOXO1) and endometrial cancer (e.g., ERRFI1, NRIP1, and EPAS1). Functional assays showed that ESR1 promotes cell viability and, in the presence of E2, enhances migration.

Conclusion

The CRISPR-mediated ESR1 activation restores estrogen responsiveness in endometrial stromal cells. Combined transcriptomic, cistromic, and chromatin architecture analyses reveal ESR1’s role in regulating decidualization and inflammation-related gene networks, with relevance to endometrial pathologies including endometrial cancer. This model serves as a powerful tool to study estrogen signaling in endometrial stromal cell biology and related pathologies.
目的:探讨ESR1基因在人子宫内膜基质细胞中的表达及雌激素驱动的转录。设计:通过crispr介导的ESR1激活,对端粒酶永生化的人子宫内膜基质细胞进行RNA-seq和Cut&Run。在原发性子宫内膜间质细胞中进行了基于hi - c的染色质结构分析(H3K27ac HiChIP)。研究对象:两名健康的育龄志愿者,月经周期规律,无妇科恶性肿瘤史。暴露:esr1激活和对照的子宫内膜基质细胞用雌二醇(E2)或对照物处理。原发子宫内膜间质细胞用载体或去个体化混合物处理。主要结果测量:差异基因表达分析(RNA-seq)鉴定了与配体无关和依赖的ESR1活性。Cut&Run分析了ESR1激活细胞中的ESR1基因组结合。H3K27ac HiChIP绘制了原代细胞中激素诱导的染色质环变化。结果:在7个测试的引导rna (gRNA)中,ESR1-3 gRNA诱导了ESR1的强大激活并恢复了E2的响应性。Bulk RNA-seq揭示了配体依赖性和非依赖性ESR1转录程序调节炎症、增殖和癌症相关途径。值得注意的是,在增殖性雌激素优势期,72%的差异表达基因与人子宫内膜组织中活跃的基因重叠,支持它们的生理相关性。Cut&Run-seq鉴定了全基因组ESR1结合位点,大多数结合位点位于远端调控元件。将Cut&Run数据与H3K27ac HiChIP染色质环结合,将远端ESR1结合位点与基因启动子连接,包括与去个体化相关的基因(如FOXO1)和子宫内膜癌(如ERRFI1、NRIP1、EPAS1)。功能分析显示ESR1促进细胞活力,并在E2存在下增强迁移。结论:crispr介导的ESR1激活可恢复子宫内膜基质细胞的雌激素反应性。转录组学、胞浆学和染色质结构的综合分析揭示了ESR1在调节脱个体化和炎症相关基因网络中的作用,并与子宫内膜癌等子宫内膜病理相关。该模型为研究雌激素信号在子宫内膜间质细胞生物学和相关病理中的作用提供了有力的工具。
{"title":"Deciphering estrogen receptor alpha–driven transcription in human endometrial stromal cells via transcriptome, cistrome, and integration with chromatin landscape","authors":"Skylar G. Montague Redecke B.Sc. ,&nbsp;Austin Bell-Hensley Ph.D. ,&nbsp;Shuyun Li Ph.D. ,&nbsp;MyeongJin Yi Ph.D. ,&nbsp;Akshadha Jain ,&nbsp;Abdull J. Massri Ph.D. ,&nbsp;Francesco J. DeMayo Ph.D.","doi":"10.1016/j.xfss.2025.08.002","DOIUrl":"10.1016/j.xfss.2025.08.002","url":null,"abstract":"<div><h3>Objective</h3><div>To investigate estrogen receptor gene 1 (ESR1) and estrogen-driven transcription in human endometrial stromal cells.</div></div><div><h3>Design</h3><div>RNA sequencing (RNA-seq) and Cleavage Under Targets and Release Using Nuclease (Cut&amp;Run) were performed on telomerase-immortalized human endometrial stromal cells with Clustered Regularly Interspaced Short Palindromic Repeats–mediated ESR1 activation. Hi-C-based chromatin architecture analysis (H3K27ac HiChIP) was conducted in primary endometrial stromal cells.</div></div><div><h3>Subjects</h3><div>Biopsies from two healthy, reproductive-aged volunteers with regular menstrual cycles and no history of gynecological malignancies.</div></div><div><h3>Exposure</h3><div>The ESR1-activated and control endometrial stromal cells were treated with estradiol (E2) or vehicle. Primary endometrial stromal cells were treated with vehicle or a decidualization cocktail.</div></div><div><h3>Main Outcome Measures</h3><div>Differential gene expression analysis (RNA-seq) identified ligand-independent and -dependent ESR1 activity. Cut&amp;Run profiled ESR1 genomic binding in ESR1-activated cells. H3K27ac HiChIP mapped hormone-induced changes in chromatin looping in primary cells.</div></div><div><h3>Results</h3><div>Among seven tested guide RNAs (gRNA), the ESR1-3 gRNA induced robust ESR1 activation and restored E2 responsiveness. Bulk RNA-seq revealed both ligand-dependent and -independent ESR1 transcriptional programs regulating inflammation, proliferation, and cancer-related pathways. Notably, 72% of differentially expressed genes overlapped with genes active in human endometrial tissue during the proliferative estrogen-dominant phase, supporting their physiological relevance. The Cut&amp;Run-seq identified genome-wide ESR1 binding sites, with most binding sites located at distal regulatory elements. Integration of Cut&amp;Run data with H3K27ac HiChIP chromatin loops linked distal ESR1 binding sites to gene promoters, including genes involved in decidualization (e.g., <em>FOXO1</em>) and endometrial cancer (e.g., <em>ERRFI1</em>, <em>NRIP1</em>, and <em>EPAS1</em>). Functional assays showed that ESR1 promotes cell viability and, in the presence of E2, enhances migration.</div></div><div><h3>Conclusion</h3><div>The CRISPR-mediated ESR1 activation restores estrogen responsiveness in endometrial stromal cells. Combined transcriptomic, cistromic, and chromatin architecture analyses reveal ESR1’s role in regulating decidualization and inflammation-related gene networks, with relevance to endometrial pathologies including endometrial cancer. This model serves as a powerful tool to study estrogen signaling in endometrial stromal cell biology and related pathologies.</div></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"7 1","pages":"Pages 74-90"},"PeriodicalIF":0.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144849965","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
Euploid but failing to implant? Insights from trophectoderm transcriptomics of euploid blastocysts using an in vitro model 整倍体但未能植入?利用体外模型观察整倍体囊胚的滋养外胚层转录组学。
Pub Date : 2026-01-01 DOI: 10.1016/j.xfss.2025.09.003
David Ortega-Jaén M.Sc. , Antonio Capalbo Ph.D. , Ángel Martín Ph.D. , Julia Gil M.Sc. , María Luisa Pardiñas M.Sc. , Carlos Mora-Martinez Ph.D. , Mireia Boluda-Navarro Ph.D. , Amparo Mercader Ph.D. , María José de los Santos Ph.D.

Objective

To investigate transcriptomic differences in mural trophectoderm (TE) cells of euploid human blastocysts on the basis of their ability to remain attached in extended in vitro culture and to identify gene expression profiles associated with competence for implantation.

Design

Prospective in vitro experimental cohort study involving ribonucleic acid sequencing of mural TE biopsies from euploid blastocysts and extended culture up to day 11.

Subjects

Fifteen euploid blastocysts donated by 16 couples undergoing intracytoplasmic sperm injection and preimplantation genetic testing for aneuploidy. Transcriptomic comparison focused on 10 euploid blastocysts, classified as attached (n = 5) or unattached (n = 5) after extended in vitro culture, and the rest were excluded for various reasons.

Exposure

Development outcome of euploid blastocysts during extended in vitro culture (adhesion vs. non-adhesion to the culture surface on day 11), used as the basis for differential gene expression analysis.

Main Outcome Measures

Identification of differentially expressed genes and deregulated molecular pathways between attached and unattached euploid embryos, including functional enrichment analysis to explore their potential roles in embryo viability and implantation.

Results

Transcriptomic analysis revealed 85 differentially expressed genes between unattached and attached euploid blastocysts, including genes involved in cellular adhesion (e.g., SPON2 and VTN), immune modulation (HLA-G and IL12A), metabolism (NNMT and CYP3A7), and mitochondrial function. Unattached embryos displayed down-regulation of ribosome biogenesis, mitochondrial proteins, and chromosomal segregation pathways and up-regulation of genes related to immune response and cytoskeletal organization.

Conclusion

The study demonstrates that mural TE gene expression differs significantly between euploid blastocysts that attach and those that do not, highlighting molecular pathways potentially linked to implantation success. Despite human implantation typically occurring through the polar TE, mural TE transcriptomic profiles may provide predictive value for embryo selection and deepen our understanding of early implantation biology. Further validation through proteomic studies and larger cohorts is needed to confirm these candidate biomarkers.
目的:研究整倍体人胚泡壁养外胚层(TE)细胞在体外培养中保持附着能力的转录组学差异,并鉴定与着床能力相关的基因表达谱。设计:前瞻性体外实验队列研究,涉及整倍体囊胚壁TE活检的RNA测序和延长培养至第11天。患者:16对夫妇捐赠的15个整倍体囊胚接受卵胞浆内单精子注射(ICSI)和着床前非整倍体基因检测(PGT-A)。转录组学比较集中在10个整倍体囊胚上,在体外延长培养后分为附着囊胚(n=5)和未附着囊胚(n=5),其余囊胚因各种原因被排除。主要观察指标:鉴定附着和未附着整倍体胚胎之间的差异表达基因(DEGs)和解除调控的分子通路,包括功能富集分析,以探讨其在胚胎存活和着床中的潜在作用。结果:转录组学分析显示,未附着和附着的整倍体囊胚之间存在85个deg,包括参与细胞粘附(如SPON2、VTN)、免疫调节(HLA-G、IL12A)、代谢(NNMT、CYP3A7)和线粒体功能的基因。未附着的胚胎显示核糖体生物发生、线粒体蛋白和染色体分离途径下调,与免疫反应和细胞骨架组织相关的基因上调。结论:该研究表明,壁TE基因表达在附着和未附着的整倍体囊胚之间存在显著差异,突出了与着床成功相关的潜在分子途径。尽管人类胚胎植入通常是通过极性TE进行的,但壁TE转录组谱可能为胚胎选择提供预测价值,并加深我们对早期植入生物学的理解。需要通过蛋白质组学研究和更大的队列进一步验证来确认这些候选生物标志物。
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引用次数: 0
Transvaginal shear wave elastography for measuring uterine myometrial and leiomyoma stiffness: a protocol pilot study 经阴道横波弹性成像测量子宫肌瘤和平滑肌瘤硬度:一项方案试点研究。
Pub Date : 2026-01-01 DOI: 10.1016/j.xfss.2025.09.002
Hannah T. Ryles M.D. , Sumeyra Agambayev M.S. , Mervat Omran Ph.D. , Chuanhong Liao M.S. , Samar Alkhrait M.D. , Ayman Al-Hendy M.D. , Ryan Longman M.D. , Jacques Abramowicz M.D. , Obianuju Sandra Madueke-Laveaux M.D.

Objective

Recent studies suggest that the stiffness of uterine leiomyomas may be related to their growth and behavior. Shear wave elastography (SWE), a quantitative method of measuring tissue stiffness, has been increasingly studied for use in gynecologic conditions. However, no protocols have been proposed for its use in this clinical setting. This study aimed to establish a standardized protocol for transvaginal SWE to measure uterine myometrial and leiomyoma stiffness and assess the reproducibility and reliability of SWE measurements. We also assessed myometrial vs. leiomyoma stiffness, compared myometrial stiffness in participants with and without leiomyomas, and assessed menstrual phase effects on stiffness values.

Design

Transvaginal SWE ultrasound measurements of myometrial and leiomyoma stiffness were obtained. All transvaginal SWE examinations were performed by an individual sonographer. Independent raters calculated stiffness measurements. Myometrial and leiomyoma stiffness were compared across menstrual phases.

Subjects

Twenty-seven premenopausal women, 16 with leiomyomas and 11 without, were enrolled. Seventeen participants completed examinations during multiple menstrual phases.

Exposure

Tissue type (myometrium/leiomyoma), leiomyoma status (presence/absence of leiomyomas), and menstrual phase (follicular/luteal).

Main Outcome Measures

Interrater and test-retest reliability of SWE measurements. Myometrial and leiomyoma shear wave values.

Results

We successfully designed a streamlined protocol for obtaining SWE measurements in participants with and without leiomyomas. Fifty-one myometrial and 38 leiomyoma stiffness values measured by 2 independent raters achieved excellent interrater reliability: the intraclass correlation coefficients between the 2 raters were 0.990 and 0.994, respectively. Fifty myometrial and 42 leiomyoma stiffness measurements calculated by a single rater at 2 time points at least 90 days apart achieved excellent test-retest reliability: intraclass correlation coefficients of 0.992 and 0.995. The median stiffness values for leiomyomas were significantly higher than those for the surrounding myometrium (48.1 [interquartile range, 39.7–59.5] vs. 31.6 [interquartile range, 22.9–46.9] kPa). There was no significant change in myometrial or leiomyoma median stiffness values between the menstrual phases.

Conclusion

Transvaginal ultrasound SWE showed excellent reproducibility and reliability in measuring myometrial and leiomyoma stiffness. Leiomyoma stiffness was significantly higher than that for the surrounding myometrium. Together, these findings support SWE’s potential clinical utility in leiomyoma management.
目的:最近的研究表明子宫平滑肌瘤的硬度可能与其生长和行为有关。横波弹性成像是一种测量组织刚度的定量方法,在妇科疾病中的应用研究越来越多。然而,在这种临床环境中,没有提出任何方案。本研究旨在1)建立经阴道剪切波弹性成像测量子宫肌瘤和平滑肌瘤硬度的标准化方案;2)评估剪切波弹性成像测量结果的可重复性和可靠性。我们还评估了子宫肌瘤与平滑肌瘤的硬度,比较了有和没有平滑肌瘤的参与者的子宫肌瘤硬度,并评估了月经期对硬度值的影响。设计:经阴道SWE超声测量子宫肌瘤和平滑肌瘤的硬度。所有经阴道SWE检查均由单个超声医师进行。独立评级计算刚度测量。子宫肌瘤和平滑肌瘤僵硬度在月经期间进行比较。受试者:27名绝经前妇女,16名有平滑肌瘤,11名无平滑肌瘤。17名参与者在多个月经期完成了测试。暴露(s): 1。组织类型(子宫肌瘤/平滑肌瘤)平滑肌瘤状态(有无平滑肌瘤)月经期(卵泡/黄体)主要结局指标:横波弹性成像测量的间测和重测可靠性。子宫肌瘤和平滑肌瘤横波值。结果:我们成功地设计了一种简化的方案,用于在有和没有平滑肌瘤的参与者中获得剪切波弹性成像测量。结果表明:经阴道超声剪切波弹性成像测量子宫肌瘤和平滑肌瘤硬度值具有良好的重现性和可靠性,其中51个子宫肌瘤硬度值和38个子宫肌瘤硬度值具有良好的组间信度,组内相关系数为0.990。平滑肌瘤的僵硬度明显高于周围的肌层。总之,这些发现支持剪切波弹性成像在平滑肌瘤治疗中的潜在临床应用。
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引用次数: 0
Enhanced sperm cryopreservation using zinc- and copper-doped hydroxyapatite with glutathione and raffinose: effects on motility, viability, deoxyribonucleic acid integrity, and oxidative stress markers 添加谷胱甘肽和棉子糖的锌和铜掺杂羟基磷灰石增强精子低温保存:对精子活力、活力、脱氧核糖核酸完整性和氧化应激标志物的影响
Pub Date : 2026-01-01 DOI: 10.1016/j.xfss.2025.10.005
Asma Mahmoudi D.V.M. , Mazdak Razi Ph.D. , Marzieh Jalilpour Ph.D. , Ali Shalizar Jalali Ph.D.

Objective

To study the protective effects of zinc- and copper-doped hydroxyapatite (H) combined with glutathione (GSH) and raffinose (R) on mouse sperm motility, viability, deoxyribonucleic acid (DNA) integrity, and oxidative stress markers before and after cryopreservation.

Design

Experimental laboratory-controlled study evaluating the efficacy of combined cryoprotective formulations during sperm cryopreservation.

Subjects

Sperm samples collected from 10 healthy adult albino mice were used for in vitro cryopreservation experiments.

Intervention

Samples were divided into six groups: a control group using a commercial cryopreservation medium and five experimental groups supplemented with H, GSH, and R, either alone or in combination (H+GSH+R). All samples were cryopreserved at −196°C for 72 hours and subsequently thawed for analysis.

Main Outcome Measures

Postthaw sperm motility, viability, and DNA fragmentation were evaluated, along with biochemical markers of oxidative stress, including total antioxidant capacity, total oxidant status, lipid peroxidation, protein oxidation, and enzymatic activities of superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase.

Results

The H+GSH+R group demonstrated significantly higher motility and lower DNA fragmentation and mortality compared with the control. Total antioxidant capacity increased prefreezing, although total oxidant status was markedly reduced postthaw. Lipid and protein oxidation decreased significantly, and antioxidant enzyme activities were enhanced in the H+GSH+R group.

Conclusion

Zinc- and copper-doped H combined with GSH and R effectively improves sperm cryosurvival by enhancing antioxidant defenses and reducing oxidative damage. This formulation provides a promising cryoprotective strategy for optimizing assisted reproductive technologies.
目的研究锌和铜掺杂羟基磷灰石(H)联合谷胱甘肽(GSH)和棉子糖(R)对小鼠精子活力、活力、脱氧核糖核酸(DNA)完整性和氧化应激标志物冷冻前后的保护作用。目的:通过实验室对照研究,评价联合冷冻保护制剂在精子冷冻保存中的效果。目的:对10只健康成年白化病小鼠进行体外冷冻保存实验。干预将样品分为6组:对照组使用商业冷冻保存培养基,5个实验组添加H、GSH和R,单独或联合(H+GSH+R)。所有样品在- 196°C低温保存72小时,随后解冻分析。主要观察指标:评估解冻后精子活力、活力和DNA片段,以及氧化应激的生化指标,包括总抗氧化能力、总氧化状态、脂质过氧化、蛋白质氧化、超氧化物歧化酶、过氧化氢酶、谷胱甘肽过氧化物酶和谷胱甘肽还原酶的酶活性。结果与对照组相比,H+GSH+R组小鼠运动能力明显提高,DNA断裂率和死亡率明显降低。冷冻前总抗氧化能力增加,但解冻后总抗氧化能力明显降低。H+GSH+R组脂质和蛋白质氧化显著降低,抗氧化酶活性增强。结论锌、铜掺杂H与GSH、R联合可有效提高精子的抗氧化防御能力,减少氧化损伤,提高精子低温存活能力。该配方为优化辅助生殖技术提供了一种有前途的冷冻保护策略。
{"title":"Enhanced sperm cryopreservation using zinc- and copper-doped hydroxyapatite with glutathione and raffinose: effects on motility, viability, deoxyribonucleic acid integrity, and oxidative stress markers","authors":"Asma Mahmoudi D.V.M. ,&nbsp;Mazdak Razi Ph.D. ,&nbsp;Marzieh Jalilpour Ph.D. ,&nbsp;Ali Shalizar Jalali Ph.D.","doi":"10.1016/j.xfss.2025.10.005","DOIUrl":"10.1016/j.xfss.2025.10.005","url":null,"abstract":"<div><h3>Objective</h3><div>To study the protective effects of zinc- and copper-doped hydroxyapatite (H) combined with glutathione (GSH) and raffinose (R) on mouse sperm motility, viability, deoxyribonucleic acid (DNA) integrity, and oxidative stress markers before and after cryopreservation.</div></div><div><h3>Design</h3><div>Experimental laboratory-controlled study evaluating the efficacy of combined cryoprotective formulations during sperm cryopreservation.</div></div><div><h3>Subjects</h3><div>Sperm samples collected from 10 healthy adult albino mice were used for in vitro cryopreservation experiments.</div></div><div><h3>Intervention</h3><div>Samples were divided into six groups: a control group using a commercial cryopreservation medium and five experimental groups supplemented with H, GSH, and R, either alone or in combination (H+GSH+R). All samples were cryopreserved at −196°C for 72 hours and subsequently thawed for analysis.</div></div><div><h3>Main Outcome Measures</h3><div>Postthaw sperm motility, viability, and DNA fragmentation were evaluated, along with biochemical markers of oxidative stress, including total antioxidant capacity, total oxidant status, lipid peroxidation, protein oxidation, and enzymatic activities of superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase.</div></div><div><h3>Results</h3><div>The H+GSH+R group demonstrated significantly higher motility and lower DNA fragmentation and mortality compared with the control. Total antioxidant capacity increased prefreezing, although total oxidant status was markedly reduced postthaw. Lipid and protein oxidation decreased significantly, and antioxidant enzyme activities were enhanced in the H+GSH+R group.</div></div><div><h3>Conclusion</h3><div>Zinc- and copper-doped H combined with GSH and R effectively improves sperm cryosurvival by enhancing antioxidant defenses and reducing oxidative damage. This formulation provides a promising cryoprotective strategy for optimizing assisted reproductive technologies.</div></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"7 1","pages":"Pages 13-26"},"PeriodicalIF":0.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145915422","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}
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F&S science
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