用于冻融公猪精液的经典精液稀释剂的比较分析

Comparative Analysis of Classic Semen Extenders for Frozen-Thawed Boar Semen.

作者信息

Kong Yuting, He Mengqian, Gao Jun, Xu Jiehuan, Lu Naisheng, Wu Caifeng, Sun Lingwei, Dai Jianjun

机构信息

College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China.

Shanghai Municipal Key Laboratory of Agri-Genetics and Breeding, Institute of Animal Science and Veterinary Medicine, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China.

出版信息

Animals (Basel). 2025 Jun 26;15(13):1885. doi: 10.3390/ani15131885.

Abstract

The physiological functions of mammalian sperm, such as motility, hyperactivation, and capacitation, require substantial energy. This study investigates the effects of two classic cryopreservation extenders-TCG (tris-citrate-glucose) and LEY (lactose-egg yolk)-on the energy metabolism of frozen-thawed boar semen. By comparing the quality indicators, key metabolite levels, and the activities of critical enzymes involved in glycolysis and the tricarboxylic acid cycle, we aim to understand how these different semen extenders influence the spermatozoa vitality of frozen-thawed boar semen. Following thawing, the LEY-cryopreserved sperm demonstrated significantly elevated motility parameters (viability, VCL, VSL, and VAP) and enhanced plasma membrane and acrosomal integrity compared with the TCG group ( < 0.05), though both cryopreserved groups exhibited significantly reduced performance relative to fresh semen controls. Cryopreservation markedly reduced intracellular adenosine triphosphate (ATP), pyruvate, and acetyl coenzyme A (A-CoA) levels (fresh > LEY > TCG; < 0.05). The LEY-preserved spermatozoa retained higher activities of glycolysis-related enzymes (phosphofructokinase, PFK; pyruvate kinase, PK) compared with the TCG group, which, in turn, showed elevated lactate dehydrogenase (LDH) activity. Critically, TCG-suppressed pyruvate dehydrogenase (PDH) activity ( < 0.05) coincided with diminished A-CoA, indicating impaired mitochondrial oxidative phosphorylation. These results demonstrate LEY's superior preservation of motility and membrane stability but highlight cryodamage-induced energy metabolism dysregulation, particularly TCG's disruption of the glycolysis-TCA cycle coordination essential for spermatozoa function. In conclusion, the choice of semen extender has a significant impact on the energy metabolism and overall quality of frozen-thawed semen, highlighting the importance of optimizing cryopreservation protocols for improved spermatozoa viability and functionality.

摘要

哺乳动物精子的生理功能,如运动能力、超激活能力和获能,都需要大量能量。本研究调查了两种经典的冷冻保存稀释液——TCG(三羟甲基氨基甲烷-柠檬酸-葡萄糖)和LEY(乳糖-蛋黄)——对冻融公猪精液能量代谢的影响。通过比较质量指标、关键代谢物水平以及参与糖酵解和三羧酸循环的关键酶的活性,我们旨在了解这些不同的精液稀释液如何影响冻融公猪精液的精子活力。解冻后,与TCG组相比,LEY冷冻保存的精子表现出显著提高的运动参数(活力、曲线速度、直线速度和平均路径速度)以及增强的质膜和顶体完整性(P<0.05),尽管两个冷冻保存组相对于新鲜精液对照组均表现出显著降低的性能。冷冻保存显著降低了细胞内三磷酸腺苷(ATP)、丙酮酸和乙酰辅酶A(A-CoA)水平(新鲜>LEY>TCG;P<0.05)。与TCG组相比,LEY保存的精子保留了更高的糖酵解相关酶(磷酸果糖激酶,PFK;丙酮酸激酶,PK)活性,而TCG组则表现出乳酸脱氢酶(LDH)活性升高。至关重要的是,TCG抑制丙酮酸脱氢酶(PDH)活性(P<0.05)与A-CoA减少同时出现,表明线粒体氧化磷酸化受损。这些结果表明LEY在运动能力和膜稳定性的保存方面更具优势,但也突出了冷冻损伤引起的能量代谢失调,特别是TCG对精子功能所必需的糖酵解-三羧酸循环协调的破坏。总之,精液稀释液的选择对冻融精液的能量代谢和整体质量有显著影响,突出了优化冷冻保存方案以提高精子活力和功能的重要性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d608/12248550/207e0d18b5c4/animals-15-01885-g001.jpg

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