Reproductive biology最新文献

Presence of embryos increases uterine blood flow and affects endometrial gene expression in Holstein cows on day seven post-insemination 受精后第7天，胚胎的存在增加了子宫血流量，影响了荷斯坦奶牛子宫内膜基因的表达

IF 2.5 3区生物学 Q3 REPRODUCTIVE BIOLOGY

Reproductive biology

Pub Date : 2024-11-30 DOI: 10.1016/j.repbio.2024.100986

Dragos Scarlet , Susanne E. Ulbrich , Michael Steufmehl , Änne Honnens , Heinrich Bollwein

Superovulation treatment leads to characteristic changes in uterine perfusion during the first three weeks of pregnancy in cattle, but the origin of these changes is unknown. We investigated embryo-induced effects on uterine blood flow volume (uBFV) and pulsatility index (uPI), as well as on steroid hormone concentrations during the first seven days after conception, together with endometrial gene expression at the time of embryo flushing, in Holstein Friesian cows. Animals (n = 8) underwent (a) artificial insemination (AI), (b) superovulation followed by artificial insemination (SOV-AI), and (c) superovulation followed by sham AI (SOV-control) in a Latin square design. Doppler ultrasound examinations and blood collections were performed 12 h before AI, as well as on days 1, 3, 5, and 7 post AI. Embryo flushing followed by endometrial biopsy collection was also performed on day 7 post AI. While uBFV was higher after SOV-AI compared to AI on days −0.5, 3, 5 and 7 post AI (P < 0.05), uPI was not affected (P > 0.05). Progesterone and total estrogen concentrations were increased following superovulation at all timepoints (P < 0.05) but were not affected by the presence of embryos (P > 0.05). Compared to AI treatment, SOV-AI was associated with increased KDR and decreased NOS3 endometrial mRNA expression (both P < 0.05) on day 7 post AI. Taken together, our results demonstrate changes in uterine blood flow during the first seven days after AI, and in endometrial gene expression in response to the presence of embryos at this time.

超排卵治疗导致牛妊娠前三周子宫灌注的特征性变化，但这些变化的来源尚不清楚。我们研究了胚胎诱导对受孕后7天子宫血流量（uBFV）、脉搏指数（uPI）以及类固醇激素浓度的影响，以及胚胎冲洗时子宫内膜基因表达的影响。动物（n = 8）在拉丁方形设计中进行(a)人工授精（AI）， (b)超排卵后人工授精（SOV-AI）， (c)超排卵后假人工授精（sov控制）。术后12 h及术后第1、3、5、7天行多普勒超声检查和采血。在人工授精后第7天进行胚胎冲洗和子宫内膜活检收集。而SOV-AI后的uBFV在AI后- 0.5、3、5和7天高于AI (P <；0.05)， uPI不受影响(P >；0.05)。超排卵后各时间点孕酮和总雌激素浓度均升高(P <；0.05)，但不受胚胎存在的影响(P >；0.05)。与AI治疗相比，SOV-AI与子宫内膜KDR升高和NOS3 mRNA表达降低相关(P <；AI后第7天0.05)。综上所述，我们的研究结果表明，人工授精后的头7天子宫血流量发生了变化，子宫内膜基因表达也发生了变化，以应对此时胚胎的存在。

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引用次数: 0

Selenium triggers AMPK-mTOR pathway to modulate autophagy related to oxidative stress of sheep Leydig cells 硒触发 AMPK-mTOR 通路，调节与绵羊亮德细胞氧化应激有关的自噬作用

IF 2.5 3区生物学 Q3 REPRODUCTIVE BIOLOGY

Reproductive biology

Pub Date : 2024-11-24 DOI: 10.1016/j.repbio.2024.100973

Liang Ma , Kexin Li , Yaru Guo , Jinyu Liu , Jianing Dong , Jun Li , Youshe Ren , Lei Shi

The objective of this study was to investigate the effect of oxidative stress induced by excessive Se on autophagy of sheep Leydig cells and its underlying mechanism. Leydig cells isolated from the testis of 8-month-old sheep were purified using a discontinuous Percoll density gradient. Cells were divided into four treatment groups (0, 2.0, 4.0 and 8.0 μmol/L of Se). After treatment with Se for 48 h, cell proliferation was detected by CCK-8 assay kit. The biochemical methods were used to evaluate the antioxidant status of Leydig cells. The mRNA transcript and protein abundance related to the AMPK-mTOR pathway and autophagy were detected by real-time PCR and western blot analysis.

The results showed that the Leydig cells treated with 8.0 μmol/L Se have the lowest cell viability. The greater ROS content and lower GSH-Px activity were also observed in the Se_8.0 group. The inclusion of 2.0 μmol/L Se in the medium did not affect the autophagy of Leydig cells. However, the relative abundance of ATG5 protein and LC3II/I ratio were elevated in the Se_8.0 group. Oxidative stress induced by excessive Se (8.0 μmol/L) dramatically improved the abundance of key proteins related to AMPK-mTOR pathway and led to an increase of phosphorylated AMPK, mTOR and ULK1. Compared with the Se_8.0 group, compound C could significantly inhibit the key molecules of AMPK-mTOR signaling pathway and mitigate the autophagy of Leydig cells induced by excessive Se. These results indicate that appropriate Se (2.0 μmol/L) can enhance the viability of sheep Leydig cells. Oxidative stress caused by Se excess can induce cell autophagy via activating AMPK-mTOR signaling pathway. The existed crosstalk between autophagy and apoptosis could decide the fate of Leydig cells. This process could play a decisive role in the maintenance of normal male fertility and spermatogenesis by affecting the number of Leydig cells in testis.

本研究旨在探讨过量 Se 诱导的氧化应激对绵羊 Leydig 细胞自噬的影响及其内在机制。采用不连续 Percoll 密度梯度法纯化从 8 个月大的绵羊睾丸中分离出的 Leydig 细胞。细胞被分为四个处理组（0、2.0、4.0 和 8.0 μmol/L Se）。用 Se 处理 48 小时后，用 CCK-8 检测试剂盒检测细胞增殖。生化方法用于评估 Leydig 细胞的抗氧化状态。结果表明，用 8.0 μmol/L Se 处理的 Leydig 细胞存活率最低。Se8.0组的ROS含量更高，GSH-Px活性更低。在培养基中加入 2.0 μmol/L Se 不会影响 Leydig 细胞的自噬。然而，Se8.0 组中 ATG5 蛋白的相对丰度和 LC3II/I 比值升高。过量Se（8.0 μmol/L）诱导的氧化应激显著提高了AMPK-mTOR通路相关关键蛋白的丰度，并导致磷酸化AMPK、mTOR和ULK1的增加。与Se8.0组相比，化合物C能显著抑制AMPK-mTOR信号通路的关键分子，并缓解过量Se诱导的Lydig细胞自噬。这些结果表明，适当的 Se（2.0 μmol/L）可以提高绵羊 Leydig 细胞的活力。Se过量引起的氧化应激可通过激活AMPK-mTOR信号通路诱导细胞自噬。自噬和细胞凋亡之间存在的相互影响可能会决定 Leydig 细胞的命运。这一过程会影响睾丸中 Leydig 细胞的数量，从而在维持男性正常生育能力和精子生成过程中发挥决定性作用。

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引用次数: 0

Empagliflozin improves sperm quality in diabetic rats by lowering oxidative stress and reducing apoptosis-related genes expression Empagliflozin 可通过降低氧化应激和减少细胞凋亡相关基因的表达来改善糖尿病大鼠的精子质量。

IF 2.5 3区生物学 Q3 REPRODUCTIVE BIOLOGY

Reproductive biology

Pub Date : 2024-11-23 DOI: 10.1016/j.repbio.2024.100971

Mina Kiani, Malek Soleimani Mehranjani, Mohammad Ali Shariatzadeh

Type 2 diabetes mellitus (T2DM) negatively impacts sperm health by increasing oxidative stress. Empagliflozin, a type of sodium-glucose cotransporter 2 (SGLT2) inhibitor, not only reduces blood glucose levels in T2DM but also has antioxidant properties. This study aimed to investigate the antioxidant effects of empagliflozin on sperm quality and spermatogenesis indices in rats with T2DM.Male rats were divided into three groups (n = 6): control, diabetes mellitus (DM, induced by streptozotocin + nicotinamide), and DM treated with empagliflozin (10 mg/kg, 56 days, via gavage). Sperm parameters including sperm count, viability, motility, normal morphology, DNA integrity, maturation, tail length, and daily sperm production were analyzed. Additionally, spermatogenesis indices, Bax/Bcl2 ratio and serum levels of superoxide dismutase (SOD), total antioxidant capacity (TAC), and malondialdehyde (MDA) were measured. In the DM group, there was a significant decrease in the mean sperm parameters, SOD, TAC, and spermatogenesis indices, along with a significant increase in the percentage of single-stranded DNA (AO⁺), immature sperm nuclei (AB⁺), MDA levels, and Bax/Bcl2 ratio, compared to the control group. In the DM + empagliflozin group, there was a significant increase in the mean sperm parameters, SOD levels (P˂0.001), and spermatogenesis indices. Additionally, there was a significant decrease in the mean AB+ (P˂0.01), AO⁺, MDA levels (P˂0.001), and Bax/Bcl2 ratio compared to the DM group. The results showed that empagliflozin can mitigate the adverse effects of diabetes and improve sperm quality and increase sperm production in diabetic rats.

2 型糖尿病（T2DM）会增加氧化应激，从而对精子健康产生负面影响。Empagliflozin 是一种钠-葡萄糖共转运体 2（SGLT2）抑制剂，它不仅能降低 T2DM 患者的血糖水平，还具有抗氧化作用。雄性大鼠分为三组（n = 6）：对照组、由链脲佐菌素+烟酰胺诱导的糖尿病组（DM）和经empagliflozin（10 mg/kg，56天，灌胃）治疗的DM组。分析了精子参数，包括精子数量、存活率、活力、正常形态、DNA完整性、成熟度、尾长和每日精子产量。此外，还测定了精子发生指数、Bax/Bcl2 比率以及血清中超氧化物歧化酶（SOD）、总抗氧化能力（TAC）和丙二醛（MDA）的水平。与对照组相比，DM组的平均精子参数、SOD、TAC和精子发生指数显著下降，单链DNA（AO+）、未成熟精子核（AB+）、MDA水平和Bax/Bcl2比值显著增加。在DM+empagliflozin组中，平均精子参数、SOD水平（P˂0.001）和精子发生指数显著增加。此外，与DM组相比，AB+（P˂0.01）、AO+、MDA水平（P˂0.001）和Bax/Bcl2比值均明显下降。结果表明，empagliflozin能减轻糖尿病的不良影响，改善糖尿病大鼠的精子质量，提高精子产量。

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引用次数: 0

Protective effect of tarantula cubensis extract on ischemia-reperfusion injury in rat ovaries: Immunohistochemical, biochemical and histopathological evaluations 狼蛛提取物对大鼠卵巢缺血再灌注损伤的保护作用：免疫组织化学、生物化学和组织病理学评估。

IF 2.5 3区生物学 Q3 REPRODUCTIVE BIOLOGY

Reproductive biology

Pub Date : 2024-11-21 DOI: 10.1016/j.repbio.2024.100970

Zahid Paksoy , Sevim Beyza Ozturk Sarikaya , Nihat Yumusak

Ischemia-reperfusion (IR) injury is a common pathology affecting the ovary. This study aimed to investigate the effect of Tarantula cubensis extract (TCE) on ovarian IR injury. Forty-two female rats were used and randomly divided into 7 groups: Control, Sham, IR, Ischemia+TCE+Reperfusion, TCE+IR, Ischemia+Ethanol+Reperfusion and Ethanol+IR. The control and sham groups did not receive any treatment. All other groups were subjected to 3 h of ischemia + 3 h of reperfusion. A single dose of TCE (0.2 ml Theranekron) was administered subcutaneously either before or after ischemia in the treatment groups. In the last 2 groups, ethanol was applied before or after ischemia. After reperfusion, blood samples were collected and analyzed for biochemical parameters. Additionally, the ovarian tissue was evaluated histopathologically and immunohistochemically. The IR group exhibited more marked hyperemia, hemorrhage, and necrosis. Both treatment groups showed histopathological improvement. Moreover, the recovery rate was greater in the animals given theranekron before ischemia. IR injury caused oxidative stress in the rats. However, Theranekron did not significantly reduce the MDA levels (p > 0.05). On the other hand, SOD levels were significantly increased in the treatment groups (p < 0.01). Compared to the control and sham groups, the IL-1β, IL-6 and TNF-α levels were very high in the IR group. Conversely, the immunoreactivity of these cytokines was further reduced in the animals given theranekron before ischemia. Our findings suggest that TCE may be beneficial in the treatment of ovarian IR damage, especially when Theranekron is given before ischemia.

缺血再灌注（IR）损伤是影响卵巢的一种常见病理现象。本研究旨在探讨虎斑蛙提取物（TCE）对卵巢IR损伤的影响。42只雌性大鼠被随机分为7组：对照组、假组、IR组、缺血+TCE+再灌注组、TCE+IR组、缺血+乙醇+再灌注组和乙醇+IR组。对照组和假组未接受任何治疗。所有其他组均接受 3 小时缺血 + 3 小时再灌注。治疗组在缺血前或缺血后皮下注射单剂量 TCE（0.2 毫升 Theranekron）。最后两组在缺血前或缺血后使用乙醇。再灌注后，收集血液样本并分析生化指标。此外，还对卵巢组织进行了组织病理学和免疫组化评估。红外组表现出更明显的充血、出血和坏死。两个治疗组的组织病理学结果均有所改善。此外，缺血前服用 theranekron 的动物恢复率更高。红外损伤会对大鼠造成氧化应激。但 Theranekron 并未显著降低 MDA 水平（p > 0.05）。另一方面，治疗组的 SOD 水平明显升高（p

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引用次数: 0

Chronic psychological stress induces testicular oxidative stress affecting reproductive behavior in rats 慢性心理压力会诱发影响大鼠生殖行为的睾丸氧化应激。

IF 2.5 3区生物学 Q3 REPRODUCTIVE BIOLOGY

Reproductive biology

Pub Date : 2024-11-20 DOI: 10.1016/j.repbio.2024.100934

Tianfeng Yang , Xinye Liu , Chunyan Kang , Gonglin Hou , Yunyun Shen , Zheqi Liu

The inhibitory effect of chronic psychological stress on reproductive behavior is widely recognized since long. However, the biological mechanisms underlying these effects, especially the cellular biology of the testicular cells, have not been fully investigated. This study aimed to investigate the effects of chronic psychological stress on rat reproductive behavior and its correlation with testicular cell damage and oxidative stress. The results showed that chronic psychological stress led to a decline in the preference scores of male rats for female rats and caused damage to the testicular tissue structure. Subcellular structures were particularly affected in the chronic psychological stress rats. Furthermore, the levels of MDA, NO, and NOS in testicular cells substantially increased under chronic psychological stress conditions. In conclusion, male reproductive behavioral disorders induced by chronic psychological stress are potentially linked to oxidative damage in testicular tissue.

长期心理压力对生殖行为的抑制作用早已得到广泛认可。然而，这些影响的生物学机制，尤其是睾丸细胞的细胞生物学机制尚未得到充分研究。本研究旨在探讨慢性心理应激对大鼠生殖行为的影响及其与睾丸细胞损伤和氧化应激的相关性。结果显示，慢性心理应激导致雄性大鼠对雌性大鼠的偏好评分下降，并对睾丸组织结构造成损伤。慢性心理应激大鼠的亚细胞结构尤其受到影响。此外，在慢性心理应激条件下，睾丸细胞中的 MDA、NO 和 NOS 水平大幅升高。总之，慢性心理应激诱发的男性生殖行为障碍可能与睾丸组织的氧化损伤有关。

引用次数: 0

Unveiling Leydig cell heterogeneity and its role in male infertility: A single-cell transcriptomic study of human testicular tissue 揭示睾丸细胞异质性及其在男性不育症中的作用：人类睾丸组织的单细胞转录组研究。

IF 2.5 3区生物学 Q3 REPRODUCTIVE BIOLOGY

Reproductive biology

Pub Date : 2024-11-19 DOI: 10.1016/j.repbio.2024.100972

Liwei Zhou , Hanchao Liu , Yuming Chen , Lin Hua , Xiaolong Wu , Xintao Gao , Le Mao

Male infertility and impaired spermatogenesis are significant concerns in reproductive health, often linked to disruptions in the cellular and molecular processes within the testis. The cellular composition and transcriptional dynamics of human testicular tissue are crucial for understanding these issues. Previous studies have largely relied on bulk tissue analysis, which obscures the distinct roles and interactions of specific cell types. Here, through a comprehensive single-cell transcriptomic analysis of human testes across various developmental stages and pathological conditions, we reveal the intricate cellular heterogeneity and the molecular mechanisms underlying testicular function. Our study identifies significant disruptions in the differentiation trajectories of Germ cells in conditions such as Klinefelter syndrome (KS), AZFa deletion, and Sertoli-cell-only syndrome (SCOS). We further uncover key transcription factors and regulatory networks governing Leydig cell function, particularly those related to steroidogenesis and hormonal regulation. These findings highlight the organized yet complex cellular and molecular landscape of the testis and uncover critical pathways altered in male infertility. Collectively, our data suggest that targeted therapeutic strategies could be developed to address specific disruptions in testicular cell populations and their associated regulatory networks.

男性不育和精子生成障碍是生殖健康领域的重大问题，通常与睾丸内的细胞和分子过程紊乱有关。人类睾丸组织的细胞组成和转录动态对于了解这些问题至关重要。以往的研究主要依赖于大块组织分析，这掩盖了特定细胞类型的独特作用和相互作用。在这里，我们通过对人类睾丸在不同发育阶段和病理条件下的单细胞转录组进行全面分析，揭示了睾丸功能背后错综复杂的细胞异质性和分子机制。我们的研究发现，在克莱菲尔特综合征（KS）、AZFa缺失和纯绒毛膜细胞综合征（SCOS）等病症中，生殖细胞的分化轨迹出现了明显的中断。我们进一步揭示了支配莱德细胞功能的关键转录因子和调控网络，尤其是与类固醇生成和激素调节相关的转录因子和调控网络。这些发现凸显了睾丸有序而复杂的细胞和分子结构，并揭示了男性不育症中发生改变的关键通路。总之，我们的数据表明，可以针对睾丸细胞群及其相关调控网络的特定紊乱，开发出有针对性的治疗策略。

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引用次数: 0

Elucidating the pathophysiology of polycystic ovary syndrome: Construction and analysis of a ceRNA network in cumulus cells 阐明多囊卵巢综合征的病理生理学：构建并分析积液细胞中的ceRNA网络

IF 2.5 3区生物学 Q3 REPRODUCTIVE BIOLOGY

Reproductive biology

Pub Date : 2024-11-19 DOI: 10.1016/j.repbio.2024.100916

Jingjing Li , Li Fan , Jiajia Wei , Wenjie Huang

Polycystic Ovary Syndrome (PCOS) is a complex endocrine disorder with elusive molecular mechanisms. This study explores the competitive endogenous RNA (ceRNA) regulatory network in the cumulus cells of PCOS patients. ceRNAs are transcripts like mRNAs, miRNAs, and lncRNAs that competitively bind shared miRNAs, regulating gene expression post-transcriptionally. We analyzed mRNA, microRNA (miRNA), and long non-coding RNA (lncRNA) from two cohorts: 12 PCOS patients and 11 healthy controls (dataset GSE10946), and 5 PCOS patients and 5 healthy controls (dataset GSE72274). These microarray datasets, obtained from the Gene Expression Omnibus (GEO), helped us identify differentially expressed mRNAs, miRNAs, and lncRNAs. Our analysis revealed a significant ceRNA network, which may play a crucial role in the pathophysiology of PCOS. In this network, 5 lncRNAs, 3 miRNAs, and 36 mRNAs were identified as differentially expressed. These elements form a complex regulatory schema influencing key cellular processes related to the disease, such as cell cycle regulation and response to estrogen. The HOXA11-AS-hsa-miR-454–3p-CCND2 network emerged as a potentially valuable biomarker for PCOS diagnosis, supported by Receiver Operating Characteristic (ROC) curve analysis indicating strong predictive power. Our findings suggest that the ceRNA interactions in PCOS cumulus cells provide a deeper understanding of the disease's molecular basis and offer new avenues for therapeutic intervention. This in silico study lays the groundwork for further experimental validation of these ceRNA networks as targets for PCOS treatment.

多囊卵巢综合征（PCOS）是一种复杂的内分泌疾病，其分子机制难以捉摸。本研究探讨了多囊卵巢综合征患者积聚细胞中的竞争性内源性 RNA（ceRNA）调控网络。ceRNA 是类似于 mRNA、miRNA 和 lncRNA 的转录本，它们能竞争性地结合共享的 miRNA，从而在转录后调控基因的表达。我们分析了两个队列中的 mRNA、microRNA (miRNA) 和长非编码 RNA (lncRNA)：12名多囊卵巢综合症患者和11名健康对照者（数据集GSE10946），以及5名多囊卵巢综合症患者和5名健康对照者（数据集GSE72274）。这些从基因表达总库（GEO）获得的微阵列数据集帮助我们鉴定了差异表达的 mRNA、miRNA 和 lncRNA。我们的分析揭示了一个重要的 ceRNA 网络，它可能在多囊卵巢综合症的病理生理学中发挥着关键作用。在这个网络中，5个lncRNA、3个miRNA和36个mRNA被鉴定为差异表达。这些元素形成了一个复杂的调控模式，影响着与该疾病相关的关键细胞过程，如细胞周期调控和对雌激素的反应。HOXA11-AS-hsa-miR-454-3p-CCND2网络成为诊断多囊卵巢综合症的潜在有价值的生物标志物，接收者操作特征曲线（ROC）分析表明它具有很强的预测能力。我们的研究结果表明，多囊卵巢综合症积液细胞中的 ceRNA 相互作用有助于深入了解该疾病的分子基础，并为治疗干预提供了新途径。这项硅学研究为进一步实验验证这些作为多囊卵巢综合症治疗靶点的 ceRNA 网络奠定了基础。

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引用次数: 0

Changes in fatty acids, vitamins, cholesterol and amino acid profiles of ram semen by freeze-thawing process 冻融过程中公羊精液脂肪酸、维生素、胆固醇和氨基酸谱的变化。

IF 2.5 3区生物学 Q3 REPRODUCTIVE BIOLOGY

Reproductive biology

Pub Date : 2024-11-19 DOI: 10.1016/j.repbio.2024.100953

İbrahim Halil Güngör , Recep Hakkı Koca , Serap Dayan Cinkara , Tutku Can Acısu , Figen Erdem Erişir , Gözde Arkalı , Şeyma Özer Kaya , Mustafa Sönmez , Seyfettin Gür , Ökkeş Yılmaz , Abdurrauf Yüce , Gaffari Türk

The objective of this study was to examine the impact of freeze-thawing on the levels of oxidative stress, fatty acids, vitamins A, D, E, and K, cholesterol, and amino acids, as well as on spermatological parameters, in ram semen. Semen was collected and pooled from each of the seven rams twice a week for three weeks. The mixed semen was diluted with tris + egg yolk diluent at 38 °C (Group 38 °C) and the temperature was reduced to 5 °C (Group 5 °C). Following the glycerolization-equilibration process (Group G-E), the samples were automatically frozen in liquid nitrogen vapor at −140 °C. The semen samples were thawed 24 h after freezing (Group Frozen-Thawed, F-T). A comparison of Group 38 °C with Group F-T revealed significant differences in several parameters. Motility rates, kinematic values, percentage of membrane integrity (HOS), some PUFA levels, ∑SFA and amino acid levels were significantly lower in Group F-T. Conversely, the ratio of dead and static spermatozoa, lipid peroxidation level, some PUFA levels, ∑MUFA, vitamins A, E and cholesterol levels were significantly higher in Group F-T. The majority of these alterations were also evident in semen samples subjected to G-E treatment. In conclusion, exposure of ram semen to G-E and F-T treatments results in modifications to semen, fatty acid, vitamin, and amino acid profiles, accompanied by elevated levels of lipid peroxidation. Moreover, this study demonstrated, for the first time, that oxidative stress was induced, some amino acid levels were altered, vitamin A and E levels were increased, vitamin D and K levels were not affected, and β-sitosterol levels were decreased after freeze-thawing in ram semen.

本研究旨在考察冻融对公羊精液中氧化应激、脂肪酸、维生素 A、D、E 和 K、胆固醇、氨基酸水平以及精子学参数的影响。七只公羊每周两次采集精液并混合，连续三周。混合后的精液在 38 °C（38 °C组）下用三酸盐+蛋黄稀释液稀释，温度降至 5 °C（5 °C组）。甘油化-校准过程（G-E 组）结束后，样本自动冷冻在 -140 °C 的液氮蒸汽中。精液样本在冷冻 24 小时后解冻（冷冻-解冻组，F-T 组）。38 °C组与F-T组的比较显示，两组在多个参数上存在显著差异。F-T 组的运动率、运动值、膜完整性百分比（HOS）、某些 PUFA 含量、∑SFA 和氨基酸含量均显著低于 F-T 组。相反，死精子和静止精子的比例、脂质过氧化水平、某些 PUFA 水平、∑MUFA、维生素 A、E 和胆固醇水平在 F-T 组明显升高。这些变化在 G-E 组处理的精液样本中也很明显。总之，将公羊精液暴露于 G-E 和 F-T 处理中会导致精液、脂肪酸、维生素和氨基酸谱的改变，并伴随着脂质过氧化水平的升高。此外，本研究还首次证明了公羊精液在冻融后会诱发氧化应激，某些氨基酸水平会发生变化，维生素 A 和 E 水平会升高，维生素 D 和 K 水平不受影响，β-谷甾醇水平会降低。

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引用次数: 0

FOXD1 activates KIFC1 to modulate aerobic glycolysis and reinforce cisplatin resistance of breast cancer FOXD1 激活 KIFC1 以调节有氧糖酵解并增强乳腺癌的顺铂耐药性。

IF 2.5 3区生物学 Q3 REPRODUCTIVE BIOLOGY

Reproductive biology

Pub Date : 2024-11-14 DOI: 10.1016/j.repbio.2024.100969

Haitao Gao, Jing Wang, Jiacai Liu, Huihua Wang, Tiantian Wang, Sha Li, Lili Niu, Ya Wei

Background

Breast cancer (BC) is the most prevalent invasive malignant tumor. Cisplatin (DDP) is a prototype of platinum-based chemotherapy drugs, its resistance severely hinders its clinical application. This project intended to figure out the exact mechanism of KIFC1 in the DDP resistance of BC.

Methods

The levels of KIFC1 and FOXD1 in BC as well as their binding sites were investigated by bioinformatics analysis. The signaling pathways regulated by FOXD1 were analyzed. Chromatin immunoprecipitation (ChIP) and dual-luciferase reporter assays verified the binding relationship between the two. Through quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot (WB), we assessed the expression of FOXD1, KIFC1, and glycolysis-related genes. CCK-8 assay was applied in the determination of cell viability to assess the efficacy of DDP resistance. Extracellular acidification rate (ECAR), glucose consumption, lactate synthesis, Adenosine triphosphate (ATP) content, and oxygen consumption rate (OCR) were measured to evaluate glycolysis.

Results

FOXD1 and KIFC1 were significantly upregulated in BC, with KIFC1 being significantly enriched in the glycolysis pathway. Overexpression of KIFC1 significantly enhanced the DDP resistance of BC cells, while promoting aerobic glycolysis. Mechanistically, FOXD1 was bound to the promoter of KIFC1 to activate its transcription. Its overexpression counteracted the inhibitory effect of KIFC1 knockdown on the DDP resistance of BC cells.

Conclusion

FOXD1 activates the glycolysis pathway by upregulating KIFC1, thereby facilitating BC cells’ DDP resistance. Therefore, the FOXD1/KIFC1 axis linked the glycolysis pathway to DDP resistance and may be a promising new target for reinforcing DDP resistance in BC.

背景：乳腺癌（BC）是发病率最高的浸润性恶性肿瘤。顺铂（DDP）是铂类化疗药物的雏形，其耐药性严重阻碍了其临床应用。本项目旨在研究KIFC1在顺铂耐药过程中的作用机制：方法：通过生物信息学分析研究了 KIFC1 和 FOXD1 在 BC 中的水平及其结合位点。分析了FOXD1调控的信号通路。染色质免疫沉淀（ChIP）和双荧光素酶报告实验验证了两者之间的结合关系。通过定量反转录聚合酶链反应（qRT-PCR）和Western印迹（WB），我们评估了FOXD1、KIFC1和糖酵解相关基因的表达。CCK-8 检测法用于测定细胞活力，以评估 DDP 抗性的有效性。测定细胞外酸化率（ECAR）、葡萄糖消耗、乳酸合成、三磷酸腺苷（ATP）含量和耗氧率（OCR）以评估糖酵解：结果：FOXD1和KIFC1在BC中明显上调，其中KIFC1在糖酵解途径中明显富集。KIFC1的过表达能显著增强BC细胞对DDP的抵抗力，同时促进有氧糖酵解。从机制上讲，FOXD1与KIFC1的启动子结合，激活了其转录。结论：FOXD1能激活KIFC1的转录，而KIFC1的过表达能抵消KIFC1敲除对BC细胞DDP抗性的抑制作用：结论：FOXD1通过上调KIFC1激活糖酵解通路，从而促进BC细胞对DDP的耐药性。因此，FOXD1/KIFC1轴将糖酵解通路与DDP耐药性联系在一起，可能是增强BC细胞DDP耐药性的一个有希望的新靶点。

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引用次数: 0

Investigating the impact of extracellular vesicle addition during IVM on the fertilization rate of equine oocytes following ICSI 研究在体外受精过程中添加细胞外囊泡对 ICSI 后马卵母细胞受精率的影响。

IF 2.5 3区生物学 Q3 REPRODUCTIVE BIOLOGY

Reproductive biology

Pub Date : 2024-11-09 DOI: 10.1016/j.repbio.2024.100967

Julia Gabryś , Natalia Pietras , Wiktoria Kowal-Mierzwa , Elżbieta Karnas , Aneta Andronowska , Agnieszka Nowak , Joanna Kochan , Monika Bugno-Poniewierska

The efficacy of in vitro embryo production (IVEP) in equines is relatively limited compared to other species due to the lack of a reliable superovulation technique, limited availability of cumulus oocyte complexes (COCs), low in vitro oocyte maturation (IVM) and fertilization rates. Extracellular vesicles (EVs), which are nanoparticles involved in intercellular signaling in the ovarian environment, have shown potential as supplements to improve oocyte development during IVM. This study tested the hypothesis that EVs from small (< 20 mm) ovarian follicles could enhance fertilization rates in mares. Follicular fluid was collected postmortem, and EVs were isolated and characterized. The IVM process was conducted with or without EVs (200 µg EV protein/ml). EV internalization during IVM was examined using fluorescent labeling and confocal microscopy. Following intracytoplasmic sperm injection (ICSI), presumptive zygotes were cultured in a time-lapse system. Confocal microscopy confirmed EV internalization by COCs. Nanoparticle tracking analysis showed that obtained EVs were submicron-sized, and flow cytometry identified surface markers CD81 and CD63 on a subpopulation of EVs. Transmission electron microscopy revealed the characteristic disk shape of EV isolates. After culture, 196 oocytes (36.84 %) exhibited a first polar body and were subjected to ICSI. The EV-treated group showed a significantly higher fertilization rate (34.7 % vs. 20.2 %; P < 0.05), reduced degeneration, and increased cleavage efficiency (P < 0.1). Despite early embryonic arrest in both groups, these results suggest that follicular fluid-derived EVs could play a supportive role in equine IVF procedures.

与其他物种相比，马体外胚胎生产（IVEP）的功效相对有限，原因是缺乏可靠的超排卵技术、卵母细胞积聚体（COCs）供应有限、体外卵母细胞成熟（IVM）和受精率较低。细胞外囊泡（EVs）是参与卵巢环境中细胞间信号传导的纳米颗粒，已显示出在体外受精过程中作为改善卵母细胞发育的补充剂的潜力。本研究对来自小卵泡（小于 20 毫米）的 EVs 可提高母马受精率的假设进行了测试。卵泡液是在死后收集的，EVs被分离出来并进行了表征。在有或没有EVs（200微克EV蛋白/毫升）的情况下进行IVM过程。使用荧光标记和共聚焦显微镜检查了IVM过程中EV的内化情况。卵胞浆内精子注射（ICSI）后，在延时系统中培养推定的合子。共聚焦显微镜证实了EV被COCs内化。纳米粒子追踪分析表明，获得的EVs为亚微米大小，流式细胞术在EVs亚群上发现了表面标记CD81和CD63。透射电子显微镜显示，EV 分离物具有特征性的圆盘形状。培养后，196 个卵母细胞（36.84%）出现了第一个极体，并进行了卵胞浆内单精子显微注射。EV处理组的受精率明显更高（34.7 % vs. 20.2 %; P

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