褪黑素对蓝狐冷冻精液中精子运动力与酶水平的影响

doi:10.16431/j.cnki.1671-7236.2025.08.021

摘要/Abstract

摘要： 【目的】研究褪黑素(melatonin,MT)对蓝狐(Vulpes lagopus)精液冷冻保存过程中精子运动性及酶水平的影响，以期为MT在精液冷冻保存中的应用提供理论依据。【方法】采集9只健康雄性蓝狐精液，经质量评估合格后混合，分为6组：未冷冻的鲜精对照组和5个MT浓度(0、0.05、0.1、0.15、0.2 mmol/L)的冷冻处理组，每组3个重复。鲜精组精液经1∶9稀释后直接用于检测，各冷冻处理组精液在等量稀释液中分别添加相应浓度MT后进行冷冻处理，解冻后检测精子活率、摆动性(WOB)、平均鞭打频率(BCF)及线粒体、抗氧化酶和顶体酶的活性等指标。【结果】 MT冷冻处理组精液的部分指标均显著低于鲜精组(P＜0.05)。在运动力方面，0.05 mmol/L MT组蓝狐精子活率、WOB、BCF、平均路径速度(VAP)、平均曲线速度(VCL)和平均直线速度(VSL)均显著高于其余MT冷冻处理组(P＜0.05)。在能量代谢方面，0.05和0.1 mmol/L MT组蓝狐精子的三磷酸腺苷(ATP)含量与线粒体呼吸链复合体Ⅰ(Complex Ⅰ)活性均显著高于0和0.2 mmol/L MT组(P＜0.05)。在氧化应激方面，0.05和0.1 mmol/L MT组蓝狐精子的活性氧(ROS)活性显著低于0和0.15 mmol/L MT组(P＜0.05)，0.05 mmol/L MT组精子的超氧化歧化酶(SOD)、过氧化氢酶(CAT)和血红素加氧酶(HO)活性均显著高于0、0.15和0.2 mmol/L MT组(P＜0.05)。在受精能力方面，0.05和0.1 mmol/L MT组蓝狐精子的顶体酶和玻璃酸酶活性均显著高于其余MT冷冻处理组(P＜0.05)。【结论】本试验条件下，在蓝狐精液冷冻稀释液中添加适量MT能有效改善冷冻精子的运动性、抗氧化能力及受精能力，以0.05 mmol/L为最佳浓度。

关键词: 蓝狐; 褪黑素; 冷冻精液; 酶水平; 运动力

Abstract: 【Objective】 The aim of this study was to investigate the effects of melatonin (MT) on sperm motility and enzymatic levels of Blue foxes (Vulpes lagopus) semen during cryopreservation,so as to provide a theoretical basis for the application of melatonin in semen cryopreservation. 【Method】 Semen samples were collected from nine healthy male Blue foxes.After quality assessment,the samples were pooled and divided into six groups:A fresh semen control group (non-cryopreserved) and five melatonin (MT) concentration gradient cryopreservation treatment groups (0,0.05,0.1,0.15,and 0.2 mmol/L).The fresh semen control group was diluted at a 1∶9 ratio and directly analyzed.Each MT freezing treatment group was supplemented with the corresponding MT concentration in an equivalent dilution medium prior to cryopreservation.Post-thaw,parameters including sperm viability,wobble (WOB),beat cross frequency (BCF),and the activities of mitochondrial,antioxidant enzyme activity,and acrosomal enzyme were evaluated. 【Result】 The partial indices in MT freezing treatment group were significantly lower than those in fresh sperm group (P＜0.05).In terms of motility,the sperm viability,WOB,BCF,average path velocity (VAP),curvilinear velocity (VCL),and straight line velocity (VSL) of Blue foxes in 0.05 mmol/L MT group were significantly higher than those in other MT freezing treatment groups (P＜0.05).In terms of energy metabolism,the ATP content and Complex Ⅰ activity of of Blue foxes sperm in 0.05 and 0.1 mmol/L MT groups were significantly higher than those in 0 and 0.2 mmol/L MT groups (P＜0.05).In terms of oxidative stress,the ROS activity of Blue foxes sperm in 0.05 and 0.1 mmol/L MT groups was significantly lower than that in 0 and 0.15 mmol/L groups (P＜0.05).The activities of superoxide dismutase (SOD),catalase (CAT),and heme oxygenase (HO) of Blue foxes sperm in 0.05 mmol/L MT group were significantly higher than those in 0,0.15 and 0.2 mmol/L MT groups (P＜0.05).In terms of fertilization ability,the activities of esterase,acrosin and HAase of Blue foxes sperm in 0.05 and 0.1 mmol/L MT groups were significantly higher than those in other MT freezing treatment groups (P＜0.05). 【Conclusion】 Under the conditions of this experiment,adding an appropriate amount of MT to the frozen diluent of semen in Blue foxes could effectively improve the sperm motility,antioxidant capacity and fertilization ability of frozen sperm,with 0.05 mmol/L as the optimal concentration.

Key words: Blue foxes; melatonin; cryopreserved semen; enzyme levels; motility

中图分类号:

S865.2⁺3

尚念鹏, 史迎秋, 韩欢胜, 魏来, 彭新宇, 王纪元, 郭丰凯, 孙义乐, 盖广辉, 孙丽英. 褪黑素对蓝狐冷冻精液中精子运动力与酶水平的影响[J]. 中国畜牧兽医, 2025, 52(8): 3726-3733.

SHANG Nianpeng, SHI Yingqiu, HAN Huansheng, WEI Lai, PENG Xinyu, WANG Jiyuan, GUO Fengkai, SUN Yile, GAI Guanghui, SUN Liying. Effect of Melatonin on the Sperm Motility and Enzyme Levels of Cryopreserved Semen in Blue Foxes[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(8): 3726-3733.

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