Ahmet Eser, Ramazan Arıcı, Selin Yağcıoğlu, İzem Sandal Asiye, Nur Ersoy, Kamber Demir, Mithat Evecen, Kemal Ak
Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine, Siirt University, Siirt, Turkey.
Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine, İstanbul University-Cerrahpasa, Avcilar, İstanbul, Turkey.
Vet Med Sci. 2025 Jan;11(1):e70172. doi: 10.1002/vms3.70172.
Ram sperm are more vulnerable to freezing than those of most other farm animals. During sperm freezing, the cell membrane loses some of its cholesterol, which regulates signalling mechanisms and prevents premature capacitation. Resveratrol (RES) increases the fluidity of the cell membrane, which becomes peroxidized during freezing and reduces free radicals. In this study, the effectiveness of RES, cholesterol-loaded cyclodextrin (CLC) and their combinations in ram sperm cryopreservation were investigated. The collected semen was divided into two equal volumes: One was diluted with tris-citric acid-glucose medium (TCG) containing CLC, whereas the other was diluted with a CLC-free TCG solution. After examining motility, both groups were further divided into two equal volumes, forming the following working groups: control (no RES, no CLC); RES (20 µg/mL); CLC (2 mg CLC/120 × 10 sperm); and RES + CLC (RES 20 µg/mL + 2 mg CLC/120 × 10 sperm). These groups were diluted with media containing their respective additives. Post-thawing, the samples were analysed for motility, acrosome and membrane integrity, membrane functionality, mitochondrial activity, capacitation status, oxidative stress and DNA integrity. CLC preserved sperm total motility, acrosome and plasma membrane integrity and decreased the rate of early capacitation (p < 0.05). RES had no significant effect on sperm quality before freezing and post-thawing (p > 0.05). However, RES + CLC increased mitochondrial activity post-thawing (p < 0.05). In conclusion, CLC minimized sperm membrane damage caused by cryopreservation in ram sperm. RES alone was ineffective, and the combination of RES and CLC did not yield a positive synergistic effect on ram spermatological parameters.
与大多数其他农场动物的精子相比，公羊精子更容易受到冷冻的影响。在精子冷冻过程中，细胞膜会失去一些胆固醇，而胆固醇可调节信号传导机制并防止过早获能。白藜芦醇（RES）可增加细胞膜的流动性，细胞膜在冷冻过程中会发生过氧化并减少自由基。在本研究中，研究了RES、胆固醇负载环糊精（CLC）及其组合对公羊精子冷冻保存的有效性。收集的精液分成两等份：一份用含有CLC的三羟甲基氨基甲烷 - 柠檬酸 - 葡萄糖培养基（TCG）稀释，而另一份用不含CLC的TCG溶液稀释。在检查活力后，两组再进一步分成两等份，形成以下工作组：对照组（无RES，无CLC）；RES组（20μg/mL）；CLC组（2mg CLC/120×10⁶精子）；RES + CLC组（RES 20μg/mL + 2mg CLC/120×10⁶精子）。这些组用含有各自添加剂的培养基稀释。解冻后，分析样品的活力、顶体和膜完整性、膜功能、线粒体活性、获能状态、氧化应激和DNA完整性。CLC可保持精子的总活力、顶体和质膜完整性，并降低早期获能率（p <0.05）。RES在冷冻前和解冻后对精子质量无显著影响（p>0.05）。然而，RES + CLC解冻后增加了线粒体活性（p <0.05）。总之，CLC可将冷冻保存对公羊精子造成的膜损伤降至最低。单独使用RES无效，RES和CLC的组合对公羊精液学参数未产生积极的协同作用。
