谷氨酰胺通过激活HSF1/HSP70通路改善绵羊冻融精液质量

doi:10.11843/j.issn.0366-6964.2025.05.021

Abstract

Abstract:

This study aimed to explore the mechanism of glutamine action during the cryopreservation process of sheep sperm. Semen samples were collected from 6 healthy adult Hu sheep aged 2-4 years. Frozen-thawed sperm motility, plasma membrane and acrosome integrity, mitochondrial membrane potential, ATP content, α-ketoglutarate dehydrogenase (α-KGDH) activity, isocitrate dehydrogenase (IDH) activity, and the expression levels of HSP70, HSF1, and phosphorylated HSF1 (pHSF1) proteins were evaluated. The results demonstrated that the addition of glutamine to the freezing extender significantly improved the total motility of sheep sperm (P < 0.05). The 5 mmol·L^-1 glutamine group exhibited significantly higher frozen-thawed sperm motility, plasma membrane and acrosome integrity, mitochondrial membrane potential, ATP content, α-KGDH activity, and IDH activity compared to the control group (P < 0.05). Glutamine significantly upregulated the expression of HSP70 and pHSF1 in frozen-thawed sperm (P < 0.05). The HSF1 inhibitor DTHIB suppressed this process and significantly reduced the total motility of frozen-thawed sperm (P < 0.05). Exogenous supplementation of HSP70 protein counteracted the inhibitory effect of DTHIB and significantly alleviated the reduction in sperm total motility induced by DTHIB treatment (P < 0.05). In this study, various quality parameters of frozen-thawed semen from sheep were examined and found that glutamine could improve the cryopreservation of sheep sperm by upregulating the phosphorylation level of HSF1 protein, increasing the expression of HSP70 protein, and protecting the mitochondrial function in frozen-thawed sperm, thereby enhancing the cryopreservation effect of sheep sperm.

Key words: sperm cryopreservation, glutamine, heat shock protein 70, heat shock factor 1

CLC Number:

S826.3

ZHANG Jian, HAI Erhan, ZHANG Jianjun, LI Boyuan, ZHANG Jiaxin. Glutamine Enhances Frozen-Thawed Semen Quality in Sheep via Activation of the HSF1/HSP70 Signaling Pathway[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(5): 2219-2229.

Figures/Tables 6

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活力参数 Motility parameter	谷氨酰胺浓度/(mmol·L^-1) Gln concentration
活力参数 Motility parameter	0	2	5	10	20
总活力/% TM	59.23±3.74^c	63.87±2.13^bc	72.90±1.35^a	68.30±0.96^ab	66.50±1.20^b
前向运动/% PM	40.20±0.49^b	43.10±1.79^b	48.33±1.48^a	40.77±1.31^b	35.43±0.83^c
平均运动速率/(μm·s^-1)VAP	62.42±3.43^c	64.08±5.11^c	88.19±0.55^a	79.74±0.77^b	67.34±0.62^c
直线运动速率/(μm·s^-1)VSL	52.86±1.03^c	52.32±2.07^c	67.13±0.06^a	68.40±0.66^a	62.20±0.54^b
曲线运动速率/(μm·s^-1)VCL	104.14±1.92^c	112.96±2.30^b	152.39±1.59^a	116.64±1.00^b	86.34±1.29^d

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Glutamine Enhances Frozen-Thawed Semen Quality in Sheep via Activation of the HSF1/HSP70 Signaling Pathway

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