Effects of anti-oxidant additives on microscopic and oxidative parameters of Angora goat semen following the freeze-thawing process
Effects of anti-oxidant additives on microscopic and oxidative parameters of Angora goat semen following the freeze-thawing process
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DOI：
10.1016/j.smallrumres.2008.03.002
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发表时间：
2008-06-01
影响因子：
1.8
通讯作者：
Kizil, Meltem
中科院分区：
文献类型：
--
作者：
Atessahin, Ahmet;Bucak, Mustafa Numan;Kizil, Meltem
还原型谷胱甘肽 (GSH)、谷胱甘肽过氧化物酶 (GSH-PX)、过氧化氢酶 (CAT) 和超氧化物歧化酶 (SOD) 的抗氧化系统被描述为对抗精液中脂质过氧化 (LPO) 的防御功能机制，对于维持精子活力和活力非常重要。精子细胞的这种抗氧化能力可能不足以防止冻融过程中的LPO。因此，本研究的目的是确定不同剂量的抗氧化添加剂对山羊精液冻融后标准精液参数、脂质过氧化和抗氧化活性的影响。对从 4 只成熟安哥拉山羊获得的精液样本（人工阴道）进行评估，并在 37 摄氏度下汇集。精液样本用 Tris 基稀释剂稀释，其中含有牛磺酸 (25、50、75 mM)、海藻糖 (25、50、75 mM) 和半胱氨酸 (5、10、15 MM)，以及不含抗氧化添加剂的稀释剂（控制）再次 评价。将稀释的精液冷却至 5 摄氏度，并在 0.25 毫升法式吸管中冷冻，然后储存在液氮中。将冷冻细管在水浴（37 摄氏度）中解冻 30 秒，用于显微镜下的精子评估。在对精液质量参数进行评估后，发现与对照组相比，使用补充有抗氧化添加剂的 Tris 补充剂不会显着降低精子死亡率。山羊精液冻融后，增加牛磺酸和海藻糖的剂量会降低精子活力（P < 0.05）。在生化测定中，与对照组相比，应用牛磺酸 (75 mM) 产生的丙二醛 (MDA) 水平最低 (4.46 +/- 0.31 nmol/ml) (P < 0.001)。与接受 5 mM 半胱氨酸的组以及对照组相比，半胱氨酸浓度为 10 和 15 mM 的组（3.27 +/- 10.11 和 3.45 +/- 0.28 nmol/ml）的 GSH 水平较低（分别为 2.27 +/- 0.08 和 2.50 +/- 0.08 nmol/ml，P < 0.001）。与对照组相比，浓度为 25 和 75 mM 的牛磺酸以及增加剂量（50 和 75 mM）的海藻糖，显着增加了 GSH-PX 活性（P < 0.01）。与其他组相比，添加 10 和 15 mM 半胱氨酸后 CAT 活性的维持率更高 (P < 0.001)。当向补充剂中添加 25 mM 牛磺酸 (329.61 +/- 6.35 mg/dl) 和 10 mM (318.64 +/- 6.34 mg/dl) 半胱氨酸时，维生素 A (VitA) 水平显着高于对照组（分别为 267.34 +/- 9.68 mg/dl 和 267.34 9.68 mg/dl） （P < 0.001）。这项研究的结果为安哥拉山羊精液冷冻保存提供了一种新方法，可能有助于山羊行业这项技术的改进。由 Elsevier B.V. 出版
The anti-oxidant system of reduced glutathione (GSH), glutathione peroxidase (GSH-PX), catalase (CAT), and superoxide dismutase (SOD) has been described as a defense functioning mechanism against lipid peroxidation (LPO) in semen, and is important in maintaining sperm motility and viability. This anti-oxidant capacity of sperm cells may be insufficient in preventing LPO during the freeze-thawing process. The aim of this study was thus to determine the influence of varying doses of anti-oxidant additives on standard semen parameters, lipid peroxidation and anti-oxidant activities after the freeze-thawing of goat semen. Ejaculate samples (artificial vagina) obtained from 4 mature Angora goats were evaluated and pooled at 37 degrees C. The semen samples diluted with a Tris-based extender, containing taurine (25, 50, 75 mM), trehalose (25, 50, 75 mM), and cysteine (5, 10, 15 MM), and an extender containing no anti-oxidant additives (control) were again evaluated. Diluted semen was cooled down to 5 degrees C and frozen in 0.25 ml French straws, prior to being stored in liquid nitrogen. Frozen straws were thawed in a water bath (37 degrees C) for 30s for microscopic sperm evaluation. Upon evaluation of parameters for semen quality, the use of a Tris-based extender supplemented with anti-oxidant additives was found to cause no significant improvement in sperm mortality, when compared to the controls. Increasing doses of taurine and trehalose decreased (P < 0.05) the sperm motility following the freeze-thawing of the goat semen. In biochemical assays, the application of taurine (75 mM) produced the lowest level of malondialdehyde (MDA) (4.46 +/- 0.31 nmol/ml), compared to the controls (P < 0.001). Lower GSH levels were higher in the groups in which cysteine was included at 10 and 15 mM (3.27 +/- 10.11 and 3.45 +/- 0.28 nmol/ml)-compared to the group which received 5 mM cysteine, as well as the controls (2.27 +/- 0.08 and 2.50 +/- 0.08 nmol/ml respectively, P < 0.001). Compared to the controls, taurine at a concentration of 25 and 75 mM, and increasing doses (50 and 75 mM) of trehalose, significantly increased the GSH-PX activity (P < 0.01). The maintenance of CAT activity was demonstrated to be higher with the addition of 10 and 15 mM cysteine, compared to the other groups (P < 0.001). Vitamin A (VitA) levels were significantly higher, compared to the controls (267.34 +/- 9.68 mg/dl and 267.34 9.68 mg/dl, respectively), when 25 mM taurine (329.61 +/- 6.35 mg/dl) and 10 mM (318.64 +/- 6.34 mg/dl) cysteine was added to the extender (P < 0.001). The results of this study provide a new approach to the cryopreservation of Angora goat semen and could contribute to the improvement of this technology in the goat industry. Published by Elsevier B.V.