Theriogenology最新文献_第3页

BECN1-mediated autophagy activates the glycolytic pathway to promote lactate synthesis in Sertoli cells of Tibetan sheep becn1介导的自噬激活糖酵解途径，促进藏羊支持细胞乳酸合成

IF 2.5 2区农林科学 Q3 REPRODUCTIVE BIOLOGY

Theriogenology

Pub Date : 2026-01-17 DOI: 10.1016/j.theriogenology.2026.117838

Xingcai Qi , Yi Wu , Qiao Li , Huihui Wang , Youji Ma

As a key regulator of autophagy, Beclin 1 (BECN1) plays an important role in spermatogenesis and the maintenance of testicular microenvironment homeostasis. However, the role of BECN1 in Tibetan sheep testes and its regulatory mechanism in Sertoli cells (SCs) remain unclear. The objective of this study was to elucidate the function of BECN1 in testicular development and to investigate whether BECN1-mediated autophagy regulates lactate synthesis in SCs. We demonstrated that BECN1 is involved in testicular development in Tibetan sheep in vivo. In vitro alteration of BECN1 expression in SCs significantly affected the expression of lactate synthesis-related genes and intracellular lactate levels. Integrated RNA-seq and untargeted metabolomics revealed that BECN1 overexpression activates the glycolytic pathway, resulting in upregulation of key glycolytic genes and accumulation of related metabolites. Moreover, inhibition of the glycolytic gene hexokinase domain component 1 (HKDC1) reversed the BECN1-induced activation of glycolysis and lactate accumulation in SCs. Collectively, these results indicate that BECN1-mediated autophagy promotes lactate synthesis by regulating the glycolytic pathway in SCs. This mechanism represents an important metabolic process during testicular development in Tibetan sheep and provides a theoretical basis for understanding male reproductive function, which may guide the breeding of superior Tibetan rams.

Beclin - 1 （BECN1）作为自噬的关键调控因子，在精子发生和睾丸微环境稳态的维持中起着重要作用。然而，BECN1在藏羊睾丸中的作用及其在支持细胞（SCs）中的调控机制尚不清楚。本研究的目的是阐明BECN1在睾丸发育中的功能，并探讨BECN1介导的自噬是否调节SCs中的乳酸合成。我们在体内证明BECN1参与藏羊睾丸发育。体外实验中，SCs中BECN1表达的改变显著影响了乳酸合成相关基因的表达和细胞内乳酸水平。综合RNA-seq和非靶向代谢组学发现BECN1过表达激活糖酵解途径，导致糖酵解关键基因上调，相关代谢物积累。此外，糖酵解基因己糖激酶结构域成分1 （HKDC1）的抑制逆转了becn1诱导的SCs中糖酵解和乳酸积累的激活。综上所述，这些结果表明becn1介导的自噬通过调节SCs中的糖酵解途径促进乳酸合成。这一机制代表了藏羊睾丸发育过程中一个重要的代谢过程，为了解雄性生殖功能提供了理论依据，为优质藏公羊的选育提供指导。

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引用次数: 0

Aldosterone regulates in vitro maturation of porcine oocytes derived from small follicles via Wnt/ β-catenin 醛固酮通过Wnt/ β-catenin调控猪小卵泡卵母细胞体外成熟。

IF 2.5 2区农林科学 Q3 REPRODUCTIVE BIOLOGY

Theriogenology

Pub Date : 2026-01-16 DOI: 10.1016/j.theriogenology.2026.117836

Wen-Jie Jiang , Chang Lu , Zhong-Le Liu , Xu-Ting Song , Shi-Yu Xiao , Juan Ma , Mei-Yu Qi , Di Liu , Yu-Chang Yao

Aldosterone (ALD) is a mineralocorticoid, an active component of the renin-angiotensin system. It has been reported that the concentration of ALD in large follicles is higher than that in small follicles. However, whether it has a beneficial effect on the maturation and development of oocytes from small follicles is currently unclear. Here, we showed that ALD supplementation during in vitro maturation promotes the maturation and development of oocytes from small follicles. Specifically, we found that 1 μg/mL ALD significantly enhanced oocyte quality by promoting cumulus expansion and polar body extrusion, increasing mitochondrial membrane potential and blastocyst rate, and reducing oxidative stress and early apoptosis. Notably, ALD supplementation enhanced the maturation and development of small follicle oocytes by inhibiting the Wnt/β-catenin pathway rather than the mineralocorticoid receptor (MR). Altogether, our findings demonstrate that ALD represents a viable strategy to enhance the quality of oocytes derived from small follicles, thereby providing a scientific basis for optimizing the in vitro maturation system for porcine oocytes.

醛固酮（ALD）是一种矿物皮质激素，是肾素-血管紧张素系统的活性成分。有报道称，大卵泡中ALD的浓度高于小卵泡。然而，它是否对小卵泡卵母细胞的成熟和发育有有益的影响目前尚不清楚。在这里，我们发现在体外成熟过程中补充ALD可以促进小卵泡卵母细胞的成熟和发育。具体而言，我们发现1 μg/mL ALD通过促进卵丘扩张和极体挤压，增加线粒体膜电位和囊胚率，减少氧化应激和早期凋亡，显著提高卵母细胞质量。值得注意的是，ALD补充通过抑制Wnt/β-catenin通路而不是矿化皮质激素受体（MR）来促进小卵泡卵母细胞的成熟和发育。总之，我们的研究结果表明，ALD是一种提高小卵泡卵母细胞质量的可行策略，从而为优化猪卵母细胞体外成熟系统提供了科学依据。

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引用次数: 0

Nano-delivery system of milk-derived exosomes loaded with Forsythiaside A: Studies on bovine mammary epithelial cells and mastitis induced mice 乳源外泌体负载连翘苷A的纳米递送系统：牛乳腺上皮细胞和乳腺炎诱导小鼠的研究。

IF 2.5 2区农林科学 Q3 REPRODUCTIVE BIOLOGY

Theriogenology

Pub Date : 2026-01-14 DOI: 10.1016/j.theriogenology.2026.117831

Xiaoqing Yang , Yangyang Pan , Meng Wang , Xin Ma , Xiaohong Han , Tianhao Li , Shanshan Yang , Jianying Chang , Hanxun Liu , Sijiu Yu , Yan Cui

Bovine mammary gland health is essential for sustaining milk yield and dairy production efficiency. Currently, antimicrobial resistance resulting from antibiotic treatment of mastitis severely constrains the sustainable development of the dairy industry. Forsythoside A (FTA), a natural phenylethanoid glycoside possessing potent anti-inflammatory and antioxidant activities, exhibits considerable therapeutic potential; however, its clinical translation remains hindered by poor bioavailability. To address this limitation, a milk-derived exosome-based nano-delivery system (FTA-mExos) was engineered. The therapeutic efficacy and underlying mechanisms of FTA-mExos were systematically evaluated in both in vitro and in vivo inflammatory models established using clinically isolated Staphylococcus chromogenes (S. chromogenes). In vitro, FTA-mExos significantly reduced reactive oxygen species (ROS) and malondialdehyde (MDA) levels in S. chromogenes-infected bovine mammary epithelial cells (BMECs), while enhancing glutathione peroxidase (GSH-Px) activity and restoring mitochondrial membrane potential. Mechanistically, FTA-mExos alleviated inflammatory responses by suppressing NF-κB signaling and downregulating the expression of pro-inflammatory cytokines, including IL-6, IL-1β, and TNF-α. In vivo studies revealed that FTA-mExos upregulated the expression of tight junction proteins (ZO-1, Occludin, and Claudin-3), leading to the restoration of blood–milk barrier (BMB) integrity and attenuation of oxidative stress injury and inflammatory responses. These findings demonstrate that FTA-mExos represent a promising nano-delivery strategy for the treatment of bovine mastitis.

牛乳腺健康对维持产奶量和奶牛生产效率至关重要。目前，乳腺炎抗生素治疗产生的耐药性严重制约着乳品行业的可持续发展。连翘苷A （FTA）是一种具有抗炎和抗氧化活性的天然苯乙醇糖苷，具有相当大的治疗潜力；然而，其临床转化仍然受到生物利用度差的阻碍。为了解决这一限制，设计了一种基于牛奶衍生的外泌体的纳米递送系统（ta - mexos）。利用临床分离的变色葡萄球菌（S. chromogenes）建立的体外和体内炎症模型，系统地评估了FTA-mExos的治疗效果和潜在机制。在体外，FTA-mExos显著降低S. chromogenes感染的牛乳腺上皮细胞（BMECs）的活性氧（ROS）和丙二醛（MDA）水平，同时增强谷胱甘肽过氧化物酶（GSH-Px）活性，恢复线粒体膜电位。机制上，FTA-mExos通过抑制NF-κB信号和下调IL-6、IL-1β和TNF-α等促炎细胞因子的表达来减轻炎症反应。体内研究表明，FTA-mExos上调紧密连接蛋白（ZO-1， Occludin和Claudin-3）的表达，导致血乳屏障（BMB）完整性的恢复，氧化应激损伤和炎症反应的减弱。这些发现表明，FTA-mExos代表了一种治疗牛乳腺炎的有前途的纳米递送策略。

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引用次数: 0

Clinical efficacy and mechanistic study of intrauterine administration of E bing angong Ye for the treatment of endometritis in sows 戊丙宫腔内给药治疗母猪子宫内膜炎的临床疗效及机制研究

IF 2.5 2区农林科学 Q3 REPRODUCTIVE BIOLOGY

Theriogenology

Pub Date : 2026-01-13 DOI: 10.1016/j.theriogenology.2026.117829

Yi-Meng Fan , Xinyi Zhou , Lu Wang , Xiyu Wen , Xueyan Wang , Lingfeng Peng , Luyao Wang , Zhihui Hao

Endometritis is a prevalent postpartum disorder in sows that significantly impairs reproductive efficiency. Traditional Chinese herbal formulations have long been applied in gynecological disorders, yet their veterinary applications remain underexplored. Ebing Angong Ye (EBAGY), composed of Curcuma aromatica (turmeric) and Dryobalanops aromatica (camphor) has historically been used to alleviate inflammatory conditions. This suggested a therapeutic potential for treating sow endometritis. We evaluated the clinical efficacy, safety and pharmacological mechanisms of EBAGY in the treatment of sow endometritis. Eighty postpartum sows diagnosed with endometritis were randomly assigned to a high, medium and low-dose EBAGY groups and a positive drug control group of chlorhexidine acetate uterine infusion (n = 20). Therapeutic efficacy was assessed by clinical symptom remission, piglet weaning weight and post-weaning estrus rate. Safety was evaluated in negative control group (n = 8) at up to 5-fold clinical doses. EBAGY treatment improved reproductive performance and had no adverse impact on hematological indices or health status. GC-MS identified borneol, curzerene, curdione and germacrone as major constituents in this formulation. Network pharmacology and in silico docking experiments also linked the therapeutic efficacy to the IL-17 signaling pathway and this was confirmed by direct cytokine measurements. Molecular docking indicated that the primary active compounds in EBAGY possessed strong binding affinities to the IL-17 receptor. ELISA assays further confirmed downregulation of the inflammatory mediators IL-17A, TNF-α, IL-6 and IL-1β. 16S rRNA sequencing was also used to demonstrate restoration of the vaginal microbiota to that of the control sows (lacking disease). EBAGY is therefore a safe and effective herbal formulation for sow endometritis, exerting therapeutic actions through anti-inflammatory effects, modulation of the IL-17 signaling pathway and restoration of microbial homeostasis.

子宫内膜炎是一种常见的产后疾病，严重影响母猪的繁殖效率。传统中药制剂在妇科疾病中的应用由来已久，但其在兽医中的应用仍未得到充分的探索。莪术安宫叶（EBAGY），由姜黄和樟脑组成，历来被用于缓解炎症。这提示了治疗母猪子宫内膜炎的治疗潜力。我们评估了EBAGY治疗母猪子宫内膜炎的临床疗效、安全性和药理机制。将诊断为子宫内膜炎的产后母猪80头随机分为EBAGY高、中、低剂量组和醋酸氯己定子宫输注阳性药物对照组（n = 20）。以临床症状缓解程度、断奶仔猪体重和断奶后发情率评价治疗效果。阴性对照组（n = 8）以高达5倍临床剂量进行安全性评估。EBAGY治疗改善了生殖性能，对血液学指标或健康状况没有不利影响。GC-MS鉴定该制剂的主要成分为冰片、curzerene、curdione和germacro。网络药理学和计算机对接实验也将治疗效果与IL-17信号通路联系起来，并通过直接细胞因子测量证实了这一点。分子对接表明，EBAGY中的主要活性化合物与IL-17受体具有较强的结合亲和力。ELISA检测进一步证实炎症介质IL-17A、TNF-α、IL-6和IL-1β下调。16S rRNA测序也用于证明阴道微生物群恢复到对照母猪（无疾病）的水平。因此，EBAGY是一种安全有效的治疗母猪子宫内膜炎的草药配方，通过抗炎作用、调节IL-17信号通路和恢复微生物稳态发挥治疗作用。

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引用次数: 0

Reproductive efficiency of Bos indicus cows under different estradiol-free ovulation synchronization protocols 不同无雌二醇排卵同步方案下印度母牛的繁殖效率

IF 2.5 2区农林科学 Q3 REPRODUCTIVE BIOLOGY

Theriogenology

Pub Date : 2026-01-13 DOI: 10.1016/j.theriogenology.2026.117830

Lucas Araujo Lemos , Laís Reis Carvalho , Matheus Pedroso Vicente , Luiz Manoel Souza Simões , Leonardo Silva Fernandes do Vale , José Camisão de Souza , José Nélio de Sousa Sales

The objective was to evaluate the fertility of Bos indicus cows submitted to an ovulation synchronization protocol without estrogen (E2). In experiment 1, injectable P4 (P4i) was administered 10 days (D-10) before the TAI protocol. On D0, cows received one intravaginal P4 device and estradiol benzoate (EB; Control) or buserelin acetate (GnRH/EC and 2GnRH). On D8, devices were removed, and all cows received eCG and PGF2α. Additionally, cows received estradiol cypionate (EC) on D8 (Control and GnRH/EC) or buserelin acetate on D10 (2GnRH). In Experiment 2, cows in the Control group underwent a protocol similar to Experiment 1. Cows in the GnRH/EC, 2GnRH/48h, and 2GnRH/54h groups were pre-synchronized with an intravaginal P4 device (D-7) and received gonadorelin and an intravaginal P4 device on D0. P4 devices were removed on D8, and all cows received eCG and PGF2α. Furthermore, the GnRH/EC group received EC, and cows in the 2GnRH/48h and 2GnRH/54h groups were administered GnRH 48 h (h) after removal of the P4 device. TAI was performed 48 (2GnRH/48h) or 54h (2GnRH/54h) after removal of the P4 device. In all experiments, the follicular diameter on D8 was greater in cows that received GnRH on D0 (P ≤ 0.01) as well as a greater ovulation rate on D0 (P ≤ 0.01). Furthermore, estrus expression was greater in cows receiving EC (P = 0.001; Exp. 2). P/AI was greater in the Control group (P = 0.001; Exp. 2). In conclusion, removal of estradiol in ovulation synchronization protocols resulted in lower fertility relative to E2-based controls.

目的是评估在没有雌激素（E2）的情况下接受排卵同步方案的波斯母牛的生育能力。实验1，在TAI方案前10天（D-10）注射P4 （P4i）。在第0天，奶牛接受1个阴道内P4装置和苯甲酸雌二醇（EB；对照）或醋酸布丝林（GnRH/EC和2GnRH）。D8时，移除设备，所有奶牛均接受心电图和PGF2α检测。此外，奶牛在D8（对照和GnRH/EC）饲喂雌二醇（EC），在D10 （2GnRH）饲喂醋酸布丝林（buserelin acetate）。在试验2中，对照组奶牛采用与试验1相似的处理方案。GnRH/EC组、2GnRH/48h组和2GnRH/54h组奶牛预先同步阴道内P4装置（D-7），并在D0时接受促性腺激素和阴道内P4装置。D8取下P4装置，所有奶牛均接受心电图和PGF2α检测。GnRH/EC组给予EC， 2GnRH/48h和2GnRH/54h组奶牛在取下P4装置48h (h)后给予GnRH。取下P4装置后48 （2GnRH/48h）或54 （2GnRH/54h）行TAI。在所有试验中，D0日接受GnRH的奶牛在D8日的卵泡直径更大（P≤0.01），D0日的排卵率更高（P≤0.01）。此外，接受EC的奶牛的发情表达更高（P = 0.001; Exp. 2）。对照组P/AI更大（P = 0.001; Exp. 2）。综上所述，与e2对照相比，在排卵同步方案中去除雌二醇导致了较低的生育能力。

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引用次数: 0

Spatial differences in the ovine cervical microenvironment at the follicular and luteal phases of the oestrous cycle; Implications for sperm transport 发情周期卵泡期和黄体期绵羊宫颈微环境的空间差异对精子运输的影响

IF 2.5 2区农林科学 Q3 REPRODUCTIVE BIOLOGY

Theriogenology

Pub Date : 2026-01-13 DOI: 10.1016/j.theriogenology.2026.117828

N. Johnston , E. Bini , E. Fitzpatrick , C. Reid , S. Fair

The complex anatomy of the ovine cervix presents a significant barrier to transcervical artificial insemination, necessitating sperm deposition at the external cervical Os. This study aimed to evaluate the cervical microarchitecture to elucidate the mechanisms orchestrating sperm transport across the cervix. Cervices from crossbred ewes (n = 13) were harvested during the follicular (n = 6) and luteal (n = 7) phase, sectioned into 10 transverse segments from external to internal Os (1 = External cervical Os; 10 = Internal cervical Os), and analysed for epithelial morphology and microgroove architecture. Columnar epithelium predominated throughout the cervix, with a squamocolumnar junction localised at the external Os. Columnar cell height was significantly greater during the follicular phase (33.3 ± 1.31 μm) than the luteal phase (26.7 ± 1.21 μm; p < 0.001). Cervical microgroove surface area increased during the follicular phase (p < 0.05). Microgroove surface area, depth, and tortuosity (deviation from a straight line) increased progressively toward the internal Os and were enhanced during the follicular phase (p < 0.05). Sialic acid expression was elevated in secondary and tertiary microgrooves during the follicular phase (p < 0.001), with mucin-bound sialic acids showing spatial variation along the canal. Segment 4 exhibited the lowest sialic acid expression (p < 0.01). Reproductive phase by cervical segment interactions affected both the secondary, tertiary microgroove epithelium, and surrounding luminal region adjacent to the tertiary microgrooves (p < 0.001). This study provides a detailed characterisation of the microarchitecture of the ovine cervical canal which is critical to understanding the mechanisms orchestrating sperm transport.

绵羊子宫颈复杂的解剖结构为经宫颈人工授精提供了一个重要的障碍，需要精子在宫颈外o处沉积。本研究旨在评估子宫颈微结构，以阐明协调精子在子宫颈运输的机制。在卵泡期（n = 6）和黄体期（n = 7）采集杂交母羊（n = 13）的宫颈，从外颈到内颈（1 =外颈o； 10 =内颈o）横切成10个横切段，分析上皮形态和微槽结构。柱状上皮主要分布于整个子宫颈，鳞状-柱状连接位于子宫颈外o区。卵泡期柱状细胞高度（33.3±1.31 μm）显著高于黄体期（26.7±1.21 μm; p < 0.001）。卵泡期宫颈微沟表面积增大（p < 0.05）。微沟槽表面积、深度和弯曲度（偏离直线）向内部o逐渐增加，并在卵泡期增强（p < 0.05）。在卵泡期，唾液酸在二级和三级微槽中的表达升高（p < 0.001），黏液结合的唾液酸在管道中表现出空间差异。第4段唾液酸表达量最低（p < 0.01）。生殖期宫颈节段相互作用影响了二级、三级微槽上皮以及邻近三级微槽的周围腔区（p < 0.001）。本研究详细描述了羊宫颈管的微结构，这对理解协调精子运输的机制至关重要。

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引用次数: 0

Comparative post-breeding outcomes in jennies inseminated with cryopreserved semen re-extended in seminal plasma or treated with platelet-rich plasma 用冷冻保存的精液受精或用富血小板血浆处理的珍妮受精后的比较结果

IF 2.5 2区农林科学 Q3 REPRODUCTIVE BIOLOGY

Theriogenology

Pub Date : 2026-01-13 DOI: 10.1016/j.theriogenology.2026.117832

Carla Joseph , Holly Schwarzman , Claire Kaplan , Marcus Machado , Bruno Trevisan , Lorenzo Segabinazzi

Artificial insemination (AI) with cryopreserved semen in donkeys is challenging due to poor fertility, likely caused by an exacerbated post-breeding inflammatory response. This study evaluated the effects of frozen-thawed semen re-extension in seminal plasma (SP) and intrauterine infusion of platelet-rich plasma (PRP) on post-breeding uterine inflammation, progesterone concentrations, and fertility in jennies. A total of 68 estrous cycles from 14 fertile jennies were randomly assigned to one of five groups: insemination with frozen-thawed semen reconstituted in 7 mL of SP (SP, n = 12); insemination with frozen-thawed semen followed by intrauterine infusion of Lactated Ringer's Solution (LRS) (Control, n = 14) or autologous PRP (PRP, n = 14) 6h post-AI; insemination with fresh semen (FS, n = 14); and sham-insemination with LRS following PRP infusion 6h post-AI (PRP.only, n = 14). Uterine lavage was performed 6h post-AI in all cycles. Intrauterine fluid accumulation (IUF), endometrial neutrophil counts (PMN), corpus luteum (CL) volume, and plasma progesterone concentration were assessed multiple times before and after AI. Pregnancy diagnosis was performed on Day 14. FS and SP groups had lower neutrophil counts 6h post-AI than Control and PRP groups (P < 0.05). PRP.only cycles had the lowest neutrophil counts at 6h and 24h post-AI (P < 0.05). Neutrophil counts were similar between all groups at 48h post-AI (P > 0.05). Plasma progesterone was higher in FS cycles on Days 3 and 8 compared to Control-assigned cycles (P < 0.05), and Day 14 compared to all groups (P < 0.05). CL volume and IUF were similar across groups (P > 0.05). Pregnancy rates were higher in FS cycles (71 %) compared to all other groups (Control, 0 %; PRP, 14 %; SP, 8 %; P < 0.05). In conclusion, SP reduced post-breeding inflammation but did not improve fertility outcomes in jennies AI with cryopreserved semen, while PRP had no effect. Additionally, plasma progesterone was affected by the type of semen used for AI but not by treatments.

由于驴的生育能力差，可能是由于繁殖后炎症反应加剧造成的，因此用低温保存的精液进行人工授精（AI）是具有挑战性的。本研究评估了冷冻解冻精液在精浆中再延伸（SP）和宫内输注富血小板血浆（PRP）对育龄后子宫炎症、孕酮浓度和生育力的影响。选取14只可育雌犬68个发情周期，随机分为5组：用7 mL SP （SP, n = 12）重组的冻融精液进行人工授精；人工授精后6h，用冻融精液授精，然后宫内输注乳酸林格液（LRS）（对照组，n = 14）或自体PRP (PRP, n = 14)；新鲜精液授精（FS, n = 14）；人工智能（PRP）后6小时注射PRP后LRS假授精。只有，n = 14)。各周期术后6h进行子宫灌洗。多次评估人工智能前后宫内积液（IUF）、子宫内膜中性粒细胞计数（PMN）、黄体（CL）体积、血浆黄体酮浓度。第14天进行妊娠诊断。人工智能后6h， FS组和SP组中性粒细胞计数低于对照组和PRP组（P < 0.05）。PRP。只有周期在ai后6h和24h中性粒细胞计数最低（P < 0.05）。术后48h各组中性粒细胞计数相近（P > 0.05）。与对照组相比，FS周期第3天和第8天血浆黄体酮水平较高（P < 0.05），第14天血浆黄体酮水平与所有组相比均较高（P < 0.05）。各组间CL体积和IUF相似（P > 0.05）。与所有其他组相比，FS周期的妊娠率（71%）更高（对照组，0%；PRP, 14%； SP, 8%; P < 0.05）。综上所述，SP减少了繁殖后的炎症，但没有改善冷冻保存精液的珍妮犬的生育结果，而PRP没有影响。此外，血浆黄体酮受用于人工智能的精液类型的影响，但不受治疗方法的影响。

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引用次数: 0

Telomere lengths in blood and sperm as biomarkers of reproductive aging and semen quality in dogs 血液和精子端粒长度作为犬生殖衰老和精液质量的生物标志物。

IF 2.5 2区农林科学 Q3 REPRODUCTIVE BIOLOGY

Theriogenology

Pub Date : 2026-01-12 DOI: 10.1016/j.theriogenology.2026.117827

Chiara Del Prete , Alessandra Iannuzzi , Valentina Longobardi , Maria Pia Pasolini , Ramona Pistucci , Alfonso Calabria , Bianca Gasparrini , Natascia Cocchia

To assess the potential utility of leukocytes and spermatozoa telomere length (LTL and STL) as reproductive biomarkers, this study measured both LTL and STL and investigated their possible correlations with oxidative status and semen quality parameters in healthy breeding male dogs. Twenty-two ejaculates and blood samples were collected from dogs of various breeds. Semen was evaluated for volume, concentration, sperm motility, and kinetic parameters using a Sperm Class Analyzer. LTL and STL were quantified using Quantitative PCR (qPCR) and expressed as the relative ratio of telomere repeat copy number (T) to a single-copy gene (S), T/S ratio. Serum oxidative stress markers were assessed using d-ROMs and Biological Antioxidant Potential (BAP) tests. Correlations between STL and LTL, as well as their association with age, semen parameters and oxidative stress levels, were evaluated using Spearman analysis. LTL and STL were 0.63 ± 0.25 and 0.85 ± 0.31 and showed a strong positive correlation (P < 0.001; r_s = 0.70). Both LTL and STL were also negatively correlated with age (P < 0.05 and rs = −0.50; P < 0.001 and rs = −0.68). The cut-off age for the difference in STL and LTL was identified at 6 and 7.5 years, respectively. STL was also positively correlated with semen volume (P < 0.05; r_s = 0.63) and concentration (P < 0.05; r_s = 0.41) and negatively correlated with semen chromatin decondensation (P < 0.01; r_s = -0.71); while the LTL showed a positive correlation with sperm concentration (P < 0.05; r_s = 0.53). No correlations with oxidative markers were found. These findings support the potential use of TL as a biomarker for reproductive aging and semen quality in dogs.

为了评估白细胞和精子端粒长度（LTL和STL）作为生殖生物标志物的潜在效用，本研究测量了健康繁殖雄性犬的LTL和STL，并研究了它们与氧化状态和精液质量参数的可能相关性。从不同品种的狗身上收集了22份射精和血液样本。使用精子类分析仪评估精液的体积、浓度、精子活力和动力学参数。LTL和STL采用定量PCR （qPCR）进行定量，用端粒重复拷贝数(T)与单拷贝基因(S)的相对比值、T/S比值表示。采用d- rom和生物抗氧化潜能（BAP）试验评估血清氧化应激标志物。使用Spearman分析评估STL和LTL之间的相关性，以及它们与年龄、精液参数和氧化应激水平的关系。LTL与STL分别为0.63±0.25和0.85±0.31，呈强正相关（P = 0.70）。LTL和STL与年龄（P = 0.63）、浓度（P = 0.41）呈负相关，与精液染色质去浓缩（P = -0.71）呈负相关；LTL与精子浓度呈正相关（P = 0.53）。未发现与氧化标志物相关。这些发现支持TL作为犬生殖衰老和精液质量的生物标志物的潜在应用。

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引用次数: 0

SB216763 enhances proliferation and sustains the undifferentiated state of porcine spermatogonial cells in prolonged In vitro culture SB216763在猪精原细胞长时间体外培养中促进细胞增殖，维持未分化状态

IF 2.5 2区农林科学 Q3 REPRODUCTIVE BIOLOGY

Theriogenology

Pub Date : 2026-01-08 DOI: 10.1016/j.theriogenology.2026.117816

Benliang Zhou , Zhen Zhang , Anqin Duan , Jiaxiang Ji , Mengqi Li , Rui Chen , Xiangxing Zhu , Jianghua Shang , Xiaogan Yang , Xingwei Liang

Spermatogonial stem cells (SSCs) serve as the foundation of spermatogenesis and play a critical role in livestock fertility and genetic improvement. However, maintaining these cells in long-term culture remains challenging, especially in pigs, where preserving both proliferative capacity and the undifferentiated state has proven difficult. To overcome this limitation, we investigated SB216763 (SB), a selective GSK-3β inhibitor reported to enhance proliferation in other stem cell types, for its potential effects on porcine spermatogonial cells (pSCs). In a 9-day concentration screening (0, 1.25, 2.5, and 5 μM SB), SB treatment significantly enhanced the proliferation of pSCs. It upregulated the proliferation-related gene PCNA and the antiapoptotic gene BCL2, while downregulating key apoptosis-related genes, including BAX and Caspase-3. It also promoted the expression of DAZL, UCHL1, NANOS2, and OCT4. Furthermore, over a 55-day period, treatment with 2.5 μM SB consistently supported higher proliferation rates, a larger proportion of DAZL-positive cells, and elevated expression of undifferentiation-associated markers (NANOS2, OCT4) compared with untreated controls at each passage. Overall, our results demonstrate that SB216763 not only promotes the proliferation of pSCs but also helps maintain their germline identity, offering a feasible strategy to improve the long-term in vitro culture of porcine spermatogonial cells.

精原干细胞（SSCs）是精子发生的基础，在家畜生育和遗传改良中发挥着重要作用。然而，在长期培养中维持这些细胞仍然具有挑战性，特别是在猪中，保持增殖能力和未分化状态已被证明是困难的。为了克服这一限制，我们研究了SB216763 (SB)，一种选择性GSK-3β抑制剂，据报道可以增强其他干细胞类型的增殖，因为它对猪精原细胞（pSCs）有潜在的影响。在为期9天的浓度筛选（0、1.25、2.5和5 μM SB）中，SB处理显著增强了psc的增殖。上调增殖相关基因PCNA和抗凋亡基因BCL2，下调BAX和Caspase-3等关键凋亡相关基因。同时促进DAZL、UCHL1、NANOS2、OCT4的表达。此外，在55天的时间内，与未处理的对照组相比，2.5 μM SB处理始终支持更高的增殖率，更大比例的dazl阳性细胞，以及非分化相关标志物（NANOS2, OCT4）的表达升高。总之，我们的研究结果表明，SB216763不仅促进了psc的增殖，而且有助于维持其种系特性，为改善猪精原细胞的长期体外培养提供了一种可行的策略。

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引用次数: 0

Oleanolic acid prevents ferroptosis and enhances oocyte competence during in vitro maturation in a porcine model 齐墩果酸在猪模型的体外成熟过程中防止铁下垂并提高卵母细胞的能力

IF 2.5 2区农林科学 Q3 REPRODUCTIVE BIOLOGY

Theriogenology

Pub Date : 2026-01-08 DOI: 10.1016/j.theriogenology.2026.117817

Xiang Zhang , Kukbin Ji , Hoyong Choi , Jaehyeok Yoon , Junghui Jo , Seonga Park , Chuang Li , Ye Liu , Inchul Choi , Min Kyu Kim

The in vitro production of porcine embryos is a valuable model for livestock biotechnology and human reproductive research. However, oocyte quality is often compromised during culture by ferroptosis, which is an iron-dependent form of cell death driven by lipid peroxidation and redox imbalance. This study induced ferroptosis using RSL3 during porcine oocyte maturation to evaluate the protective potential of oleanolic acid (OA), an antioxidant triterpenoid compound. Exposure to RSL3 increased Fe²⁺ accumulation (as detected by FerroOrange staining), lipid peroxidation (as determined by the MDA assay), apoptosis, and mitochondrial dysfunction. These changes were consistent with ferroptotic stress and were accompanied by impaired cumulus expansion, meiotic progression, and blastocyst formation. Supplementing with 1 mg/L OA significantly improved oocyte competence, partially restored blastocyst development, enhanced cumulus expansion, and reduced apoptosis. At the cellular level, OA reduced ROS, MDA, and abnormal mitochondrial distribution while maintaining glutathione levels and mitochondrial membrane potential. Molecular analysis revealed that OA modulated ferroptosis-related gene expression, including partial restoration of GPX4 and SLC7A11 expression and suppression of ACSL4 and TFRC upregulation. Collectively, these findings indicate that OA partially alleviated RSL3-induced ferroptotic damage in porcine oocytes, as reflected by coordinated improvements in iron homeostasis, redox status, mitochondrial function, and the expression of ferroptosis- and autophagy-associated markers. OA could therefore represent a promising technique to improve oocyte quality and embryonic development, with potential application value in reproductive medicine and livestock biotechnology.

猪胚胎的体外生产是家畜生物技术和人类生殖研究的一个有价值的模式。然而，在培养过程中，卵母细胞质量经常受到铁凋亡的影响，铁凋亡是一种由脂质过氧化和氧化还原失衡驱动的铁依赖性细胞死亡形式。本研究利用RSL3诱导猪卵母细胞成熟过程中的铁凋亡，以评估齐齐果酸（OA）（一种抗氧化的三萜化合物）的保护潜力。暴露于RSL3会增加Fe2+积累（通过铁橙色染色检测）、脂质过氧化（通过MDA测定）、细胞凋亡和线粒体功能障碍。这些变化与紧铁应激一致，并伴有积云扩张、减数分裂进程和囊胚形成受损。添加1 mg/L OA可显著提高卵母细胞能力，部分恢复囊胚发育，增强卵丘扩张，减少细胞凋亡。在细胞水平上，OA降低ROS、MDA和线粒体异常分布，同时维持谷胱甘肽水平和线粒体膜电位。分子分析表明，OA调节了铁中毒相关基因的表达，包括部分恢复GPX4和SLC7A11的表达，抑制ACSL4和TFRC的上调。综上所述，这些发现表明OA部分缓解了rsl3诱导的猪卵母细胞铁凋亡损伤，这可以通过铁稳态、氧化还原状态、线粒体功能以及铁凋亡和自噬相关标记物表达的协同改善来反映。因此，OA在提高卵母细胞质量和胚胎发育方面是一种很有前景的技术，在生殖医学和家畜生物技术方面具有潜在的应用价值。

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引用次数: 0