Sun Linlin, Fan Xiaoteng, Zeng Yao, Wang Liqian, Zhu Zhendong, Li Rongnan, Tian Xiue, Wang Yongjun, Lin Yan, Wu De, Zeng Wenxian
Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, China.
Key Laboratory for Animal Disease Resistance Nutrition of the Ministry of Education of China, Institute of Animal Nutrition, Sichuan Agricultural University, Ya'an, China.
Zygote. 2020 Jun 2:1-8. doi: 10.1017/S0967199420000271.
The objective of the present study was to elucidate whether resveratrol could facilitate the survival of boar sperm during liquid preservation and fast cooling processes. Boar semen were diluted with Modena extender containing different concentrations of resveratrol. Sperm motility was evaluated by visual estimation. Membrane integrity, acrosome integrity and mitochondrial membrane potentials were measured by SYBR-14/PI, FITC-PNA and JC-1 staining, respectively. Moreover, the levels of reactive oxygen species (ROS), malonaldehyde (MDA) and total antioxidant capacity (T-AOC) were measured using commercial assay kits. B-cell lymphoma protein-2 (BCL2) content was determined by western blotting. During liquid preservation at 17oC, the addition of 50 μM resveratrol to the Modena extender significantly improved sperm motility, membrane integrity, acrosome integrity, and sperm mitochondrial membrane potentials. Similar results were also observed in the 150 μM resveratrol group during the fast cooling process. Furthermore, addition of resveratrol led to a decrease of ROS and MDA, and an increase in the content of T-AOC and BCL2. These observations suggest that addition of resveratrol to Modena extender protects boar sperm against oxidative stress. The optimal concentrations of resveratrol are 50 μM and 150 μM during liquid preservation and fast cooling process, respectively.
本研究的目的是阐明白藜芦醇是否能在公猪精液液体保存和快速冷却过程中促进精子存活。用含有不同浓度白藜芦醇的摩德纳稀释液对公猪精液进行稀释。通过视觉评估来评价精子活力。分别用SYBR-14/PI、FITC-PNA和JC-1染色法测定膜完整性、顶体完整性和线粒体膜电位。此外,使用商业检测试剂盒测量活性氧(ROS)、丙二醛(MDA)和总抗氧化能力(T-AOC)的水平。通过蛋白质印迹法测定B细胞淋巴瘤蛋白-2(BCL2)的含量。在17℃液体保存期间,向摩德纳稀释液中添加50μM白藜芦醇可显著提高精子活力、膜完整性、顶体完整性和精子线粒体膜电位。在快速冷却过程中,150μM白藜芦醇组也观察到类似结果。此外,添加白藜芦醇可导致ROS和MDA含量降低,T-AOC和BCL2含量增加。这些观察结果表明,向摩德纳稀释液中添加白藜芦醇可保护公猪精子免受氧化应激。在液体保存和快速冷却过程中,白藜芦醇的最佳浓度分别为50μM和150μM。