矢车菊素-3-芸香糖苷对奶绵羊精液冷冻保存效果的影响

doi:10.11843/j.issn.0366-6964.2025.04.025

摘要/Abstract

摘要：

旨在探究稀释液中添加矢车菊素-3-芸香糖苷(cyanidin-3-rutinoside，C3R)对奶绵羊精液冷冻保存效果的影响，并研究其作用机理。本研究采集6只年龄2.5~3岁且体况良好的东佛里生种公羊的新鲜精液，镜检合格后混匀并分为6组，其中1组是用不含C3R的稀释液仅作稀释处理的冻前组，其余5组分别用不同浓度C3R(0、20、40、60、80 μg·mL^-1)的冷冻稀释液对精液进行稀释及冷冻保存，每组3个重复。通过检测解冻后精子质量、抗氧化能力以及腺苷代谢相关指标，探究C3R对奶绵羊精液冷冻保存效果的影响。结果表明：C3R的添加显著提高了精子质量，其中40 μg·mL^-1 C3R处理组的精子前向运动比例和顶体完整率最高，显著高于对照组(P < 0.05)；60 μg·mL^-1 C3R处理组的质膜完整率最高，显著高于对照组(P < 0.05)。C3R的添加显著提高了精子抗氧化能力，其中40 μg·mL^-1 C3R处理组精子的超氧化物歧化酶活性显著高于其他组(P < 0.05)，丙二醛含量显著低于其他组(P < 0.05)；60 μg·mL^-1 C3R处理组的总抗氧化能力和过氧化氢酶活性显著高于其他组(P < 0.05)；20、40 μg·mL^-1处理组的活性氧含量显著低于对照组(P < 0.05)。综上，在本试验条件下40 μg·mL^-1 C3R对奶绵羊精子的保护效果最佳。在腺苷代谢方面，冻后组精子的腺苷脱氨酶活性显著高于冻前组(P < 0.05)，而40 μg·mL^-1 C3R的添加显著抑制了腺苷脱氨酶的活性(P < 0.05)，同时显著降低了腺苷和肌苷的含量(P < 0.05)。因此，在冷冻稀释液添加C3R能够提高奶绵羊冷冻精液保存效果。当C3R添加浓度为40 μg·mL^-1时，能够显著提高精子的抗氧化能力并降低腺苷脱氨酶活性，从而提高了奶绵羊冻后精液的质量。

关键词: 奶绵羊, 精液, 冷冻保存, 矢车菊素-3-芸香糖苷, 抗氧化, 腺苷脱氨酶

Abstract:

This study aimed to investigate the effect of cyanidin-3-rutinoside added to the diluent on the cryopreservation of dairy sheep semen and to explore its mechanism of action. Fresh semen was collected from 6 East Friensian rams aged 2.5 to 3 years old and in good physical condition. After undergoing microscopic examination, the semen was mixed and divided into 6 groups. One group served as the pre-freezing group, which was diluted with a diluent without C3R. The remaining 5 groups were diluted and cryopreserved using diluents containing varying concentrations of C3R (0, 20, 40, 60, and 80 μg·mL^-1). Each group consisted of 3 replicates. The effect of C3R on the cryopreservation of dairy sheep semen was studied by evaluating related indicators of sperm quality, antioxidant capacity, and adenosine metabolism after thawing. The results indicated that the addition of C3R significantly improved sperm quality. The treatment group receiving 40 μg·mL^-1 of C3R exhibited the highest proportion of sperm forward movement and acrosome integrity, both of which were significantly higher than those in the control group (P < 0.05); The plasma membrane integrity rate in the 60 μg·mL^-1 C3R treatment group was the highest among all groups and was significantly higher than that of the control group (P < 0.05). The addition of C3R significantly enhanced the antioxidant capacity of sperm. The superoxide dismutase activity of sperm in the 40 μg·mL^-1 C3R treatment group was significantly higher than that in the other groups (P < 0.05), while the malondialdehyde content was significantly lower than that in the other groups. (P < 0.05); The total antioxidant capacity and catalase activity in the 60 μg·mL^-1 C3R treatment group were significantly higher than those in the other groups (P < 0.05); The active oxygen content in the 20 and 40 μg·mL^-1 treatment groups was significantly lower than that in the control group (P < 0.05). Therefore, under the conditions of this experiment, 40 μg·mL^-1 C3R demonstrated the best protective effect on dairy sheep sperm. Regarding adenosine metabolism, the activity of adenosine deaminase in the frozen group was significantly higher than that in the pre-frozen group (P < 0.05). Furthermore, the addition of 40 μg·mL^-1 C3R significantly inhibited the activity of adenosine deaminase and reduced the contents of adenosine and inosine (P < 0.05). Therefore, incorporating C3R into the freezing diluent can enhance the preservation efficacy of frozen semen from dairy sheep. When C3R is added at a concentration of 40 μg·mL^-1, it significantly boosts the antioxidant capacity of the sperm and decreases adenosine deaminase activity, thereby improving the quality of frozen semen from dairy sheep.

Key words: dairy sheep, semen, cryopreservation, C3R, antioxidant, adenosine deaminase

中图分类号:

S826.3

马应天, 姜璐瑶, 李增开, 秦剑平, 赵建华, 贺玉芳, 宋宇轩, 张磊. 矢车菊素-3-芸香糖苷对奶绵羊精液冷冻保存效果的影响[J]. 畜牧兽医学报, 2025, 56(4): 1768-1778.

MA Yingtian, JIANG Luyao, LI Zengkai, QIN Jianping, ZHAO Jianhua, HE Yufang, SONG Yuxuan, ZHANG Lei. Effect of Cyanidin-3-rutinoside on Cryopreservation of Semen of Dairy Sheep[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(4): 1768-1778.

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精子活力参数 Sperm motility parameter	冻前组 Before freezing	矢车菊素-3-芸香糖苷/(μg·mL^-1) C3R
精子活力参数 Sperm motility parameter	冻前组 Before freezing	0	20	40	60	80
前向运动比例/% PM	66.71±1.89^a	40.64±0.97^d	44.69±0.33^c	50.74±0.72^b	49.36±0.52^b	43.35±0.92^c
平均曲线速度/(μm·s^-1)VCL	83.24±2.38^a	58.04±7.57^c	59.61±2.16^c	75.27±7.81^ab	68.75±3.88^bc	62.39±5.69^c
平均直线速度/(μm·s^-1)VSL	26.76±0.11^a	19.37±0.67^c	19.47±2.97^c	24.08±1.59^ab	22.77±1.55^bc	21.24±2.56^bc
平均路径速度/(μm·s^-1)VAP	42.62±1.68^a	27.10±2.00^d	27.49±2.75^cd	34.32±2.74^b	31.96±0.68^bc	29.47±3.12^cd
摆动性/% WOB	42.46±0.97^a	46.93±2.75^a	46.06±3.18^a	45.68±1.54^a	46.61±3.10^a	47.20±1.54^a

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