Departamento de Clínica e Cirurgia Veterinárias da Escola de Veterinária da Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil.
Departamento de Farmacologia do Instituto de Ciências Biológicas da UFMG, Belo Horizonte, MG, Brazil.
J Equine Vet Sci. 2020 May;88:102964. doi: 10.1016/j.jevs.2020.102964. Epub 2020 Feb 13.
During semen cryopreservation, the sensitivity of equine sperm to oxidative stress is increased by the eliminated seminal plasma. Thus, antioxidant addition to the semen extender can be helpful to the sperm survival after freezing and thawing. This work aimed to test whether coenzyme Q10 (CoQ10) added in different concentrations to the INRA 82 freezing extender has antioxidant function on equine sperm to improve its fertilizing ability. Semen samples from five stallions were frozen with the extenders: (T1) INRA 82, control, (T2) T1+ 5 μM CoQ10, (T3) T1+ 25 μM CoQ10, and (T4) T1+ 50 μM CoQ10. After sample thawing, sperm motility and kinetics characteristics were evaluated using a computer-assisted sperm analysis and sperm membrane functionality and integrity were evaluated with a hypo-osmotic swelling test and an epifluorescence microscopy, respectively. The nitrite (NO) and hydrogen peroxide (HO) concentrations of the semen samples were measured with spectrophotometry. There was no difference on the sperm characteristics among all treatments (P > .05). However, the 25 μM CoQ10 (T3) decreased NO concentration (6.7 ± 2.2 μM/μg protein) compared with the treatments T1, T2, and T4 (64.3 ± 3.7, 59.4 ± 5.3, 45.1 ± 8.6 μM/μg protein), respectively, as well HO concentration (1.8 ± 0.3 μM/μg protein) compared with the control (4.6 ± 0.4 μM/μg protein) and 5 μM CoQ10 treatments (4.8 ± 0.2 μM/μg protein, P < .05). In conclusion, 25 μM CoQ10 plays a significant role as antioxidant to the frozen equine sperm, decreasing NO and HO concentrations. Thus, its addition to the INRA 82 freezing extender may be beneficial to the fertilizing ability of equine semen.
在精液冷冻保存过程中，去除精浆会增加马精子对氧化应激的敏感性。因此，在精液稀释液中添加抗氧化剂有助于冷冻和解冻后精子的存活。本研究旨在测试辅酶 Q10（CoQ10）以不同浓度添加到 INRA 82 冷冻稀释液中是否对马精子具有抗氧化作用，从而提高其受精能力。从五头种马采集精液样本，用以下稀释液冷冻：（T1）INRA 82，对照，（T2）T1+5 μM CoQ10，（T3）T1+25 μM CoQ10，和（T4）T1+50 μM CoQ10。样本解冻后，使用计算机辅助精子分析评估精子运动和动力学特征，使用低渗肿胀试验和荧光显微镜评估精子膜功能和完整性。使用分光光度法测量精液样本中的亚硝酸盐（NO）和过氧化氢（HO）浓度。所有处理之间的精子特征均无差异（P>.05）。然而，与处理 T1、T2 和 T4 相比，25 μM CoQ10（T3）降低了 NO 浓度（6.7±2.2 μM/μg 蛋白），分别为 64.3±3.7、59.4±5.3 和 45.1±8.6 μM/μg 蛋白，以及 HO 浓度（1.8±0.3 μM/μg 蛋白），与对照（4.6±0.4 μM/μg 蛋白）和 5 μM CoQ10 处理（4.8±0.2 μM/μg 蛋白）相比，P<.05。总之，25 μM CoQ10 作为抗氧化剂对冷冻马精子发挥重要作用，降低了 NO 和 HO 浓度。因此，将其添加到 INRA 82 冷冻稀释液中可能有益于马精液的受精能力。
