Effects of antioxidants on post-thawed bovine sperm and oxidative stress parameters: Antioxidants protect DNA integrity against cryodamage
Effects of antioxidants on post-thawed bovine sperm and oxidative stress parameters: Antioxidants protect DNA integrity against cryodamage
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DOI：
10.1016/j.cryobiol.2010.09.001
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发表时间：
2010-12-01
期刊：
影响因子：
2.7
通讯作者：
Oztuna, Derya
中科院分区：
文献类型：
--
作者：
Bucak, Mustafa Numan;Tuncer, Purhan Barbaros;Oztuna, Derya
本研究旨在确定冻融过程后蛋氨酸、肌醇和肉碱对牛精液精子（活力、异常、DNA 完整性和体内生育能力）和氧化应激参数（脂质过氧化、总谷胱甘肽和抗氧化电位水平）的影响。该研究包括每周两次借助人造阴道从每头西门塔尔牛身上收集九份精液。将每份精液分成七等份，并在含有蛋氨酸（2.5 和 7.5 mM）、肉碱（2.5 和 7.5 mM）、肌醇（2.5 和 7.5 mM）且无添加剂（对照）的 Tris 基增量剂中稀释，冷却至 5 摄氏度，然后冷冻在 0.25 ml 吸管中。然后将冷冻吸管在 37 摄氏度的水浴中单独解冻 20 秒进行评估。与其他组相比，添加 7.5 mM 剂量的肉碱和肌醇的补充剂导致更高的主观运动百分比（61.9 +/- 1.3% 和 51.3 +/- 1.6%）。与对照和肌醇 2.5 mM 相比，添加 2.5 和 7.5 mM 剂量的蛋氨酸和肉碱以及 7.5 mM 剂量的肌醇对总异常百分比提供了更大的保护作用（P < 0.001）。至于 CASA 活力，与其他组相比，7.5 mM 肉碱（41.6 +/- 2.9% 和 54.2 +/- 4.9%）和肌醇（34.9 +/- 2.0% 和 47.3 +/- 2.2%）引起 CASA 和总活力的不显着增加。 2.5 和 7.5 mM 的所有抗氧化剂均导致 DNA 受损的精子数量低于对照组，从而减少了 DNA 损伤（P < 0.05）。各组之间的 CASA 前向运动和精子运动特征没有观察到显着差异。在基于 59 天不返回的生育结果中，各组之间的不返回率没有观察到显着差异。在生化参数方面，与对照组相比，补充抗氧化剂并未显着影响 LPO 和总 GSH 水平（P > 0.05）。冻融后，蛋氨酸 2.5 mM 中 AOP 水平的维持率 (5.06 +/- 0.38 mM) 高于对照 (0.96 +/- 0.29 mM) (P < 0.001)。在冷冻保存过程之前补充这些抗氧化剂可以保护 DNA 完整性免受冷冻损伤。此外，未来的研究应集中于抗氧化剂蛋氨酸、肉碱和肌醇在冷冻保存过程中抗氧化作用的分子机制。 (C) 2010 Elsevier Inc. 保留所有权利。
This study was conducted to determine the effects of methionine, inositol and carnitine on sperm (motility, abnormality, DNA integrity and in vivo fertility) and oxidative stress parameters (lipid peroxidation, total glutathione and antioxidant potential levels) of bovine semen after the freeze-thawing process. Nine ejaculates, collected with the aid of an artificial vagina twice a week from each Simmental bovine, were included in the study. Each ejaculate, splitted into seven equal groups and diluted in Tris-based extender containing methionine (2.5 and 7.5 mM), carnitine (2.5 and 7.5 mM), inositol (2.5 and 7.5 mM) and no additive (control), was cooled to 5 degrees C and then frozen in 0.25 ml straws. Frozen straws were then thawed individually at 37 degrees C for 20 s in a water bath for the evaluation.The extender supplemented with 7.5 mM doses of carnitine and inositol led to higher subjective motility percentages (61.9 +/- 1.3% and 51.3 +/- 1.6%) compared to the other groups. The addition of methionine and carnitine at doses of 2.5 and 7.5 mM and inositol at doses of 7.5 mM provided a greater protective effect in the percentages of total abnormality in comparison to the control and inositol 2.5 mM (P < 0.001). As regards CASA motility, 7.5 mM carnitine (41.6 +/- 2.9% and 54.2 +/- 4.9%) and inositol (34.9 +/- 2.0% and 47.3 +/- 2.2%) caused insignificant increases in CASA and total motility in comparison to the other groups. All of the antioxidants at 2.5 and 7.5 mM resulted in lower sperm with damaged DNA than that of control, thus reducing the DNA damage (P < 0.05). No significant differences were observed in CASA progressive motility and sperm motion characteristics among the groups. In fertility results based on 59-day non-returns, no significant differences were observed in non-return rates among groups. As regards biochemical parameters, supplementation with antioxidants did not significantly affect LPO and total GSH levels in comparison to the control group (P > 0.05). The maintenance of AOP level in methionine 2.5 mM was demonstrated to be higher (5.06 +/- 0.38 mM) than that of control (0.96 +/- 0.29 mM) following the freeze-thawing (P < 0.001). Supplementation with these antioxidants prior to the cryopreservation process protected the DNA integrity against the cryodamage. Furthermore, future research should focus on the molecular mechanisms of the antioxidative effects of the antioxidants methionine, carnitine and inositol during cryopreservation. (C) 2010 Elsevier Inc. All rights reserved.