Gerzilov Vasko, Andreeva Madlena, Tsvetanova Elina, Georgieva Almira, Alexandrova Albena
Department of Animal Science, Agricultural University, Plovdiv, Bulgaria.
Laboratory of Free Radical Processes, Institute of Neurobiology, Bulgarian Academy of Sciences, Sofia, Bulgaria.
Reprod Domest Anim. 2023 Dec;58(12):1702-1711. doi: 10.1111/rda.14487. Epub 2023 Oct 11.
The aim of this study was to evaluate the effect of antioxidant supplementation in diluted semen from Muscovy drakes after the induction of oxidative stress (OS) on the sperm motility, kinematic parameters and biochemical markers - lipid peroxidation (LPO) levels and total glutathione (tGSH) concentration. The pooled semen was distributed equally into three parts, diluted (1:3 v/v) with IMV Canadyl, HIA-1 or AU, and stored at 4°C for 6 h. Later, the semen was equilibrated at 20-25°C for 15 min, and divided in Eppendorf tubes. The sperm samples (final concentration of 50 × 10 sperm cells/mL per sample) were incubated at 37°C for 30 min in the absence (-) or presence (+) of 0.1 mM FeSO + 0.5 mM H O (Fenton system) and the following combinations of antioxidants: ascorbic acid + Trolox (A + T); ascorbic acid + Desferal (A + D); Trolox + Desferal (T + D) and ascorbic acid + Trolox + Desferal (A + T + D), all of them in a final concentration of 0.1 mM. Thus, the total number of samples was 30 and in each one, the sperm motility, velocity parameters, LPO and tGSH were determined. The motility and kinematic parameters of the diluted semen with added antioxidants were restored by up to 20% after inducing OS via the Fenton reaction. Dual combinations of antioxidants (A + T, A + D, and T + D) lowered LPO levels, but not equally across different extenders. After the induction of OS, the tGSH levels in diluted semen with IMV-Canadyl were not affected by the added antioxidants. Whereas antioxidant combinations in diluted semen with HIA-1 or AU had a beneficial effect and partially restored tGSH levels. In conclusion, the results showed that the extender IMV-Canadyl is well balanced and protected the Muscovy semen under OS conditions, while the other two extenders HIA-1 and AU can be improved by adding antioxidants.
本研究的目的是评估抗氧化剂补充对番鸭稀释精液在诱导氧化应激(OS)后精子活力、运动学参数和生化标志物——脂质过氧化(LPO)水平及总谷胱甘肽(tGSH)浓度的影响。将混合精液平均分为三份,分别用IMV Canadyl、HIA - 1或AU稀释(1:3 v/v),并在4°C下保存6小时。之后,精液在20 - 25°C下平衡15分钟,然后分装到Eppendorf管中。精子样本(每个样本最终浓度为50×10个精子细胞/mL)在37°C下孵育30分钟,孵育条件为不存在( - )或存在( + )0.1 mM FeSO₄ + 0.5 mM H₂O₂(芬顿体系)以及以下抗氧化剂组合:抗坏血酸 + 生育三烯酚(A + T);抗坏血酸 + 去铁胺(A + D);生育三烯酚 + 去铁胺(T + D)和抗坏血酸 + 生育三烯酚 + 去铁胺(A + T + D),所有这些抗氧化剂的最终浓度均为0.1 mM。因此,样本总数为30个,每个样本均测定精子活力、速度参数、LPO和tGSH。通过芬顿反应诱导OS后,添加抗氧化剂的稀释精液的活力和运动学参数最多可恢复20%。抗氧化剂的二元组合(A + T、A + D和T + D)降低了LPO水平,但在不同稀释剂中的效果并不相同。诱导OS后,添加抗氧化剂对用IMV - Canadyl稀释的精液中的tGSH水平没有影响。而用HIA - 1或AU稀释的精液中的抗氧化剂组合具有有益作用,并部分恢复了tGSH水平。总之,结果表明,稀释剂IMV - Canadyl平衡性良好,在OS条件下能保护番鸭精液,而另外两种稀释剂HIA - 1和AU可通过添加抗氧化剂得到改善。