Li Shubin, Ren Jingyu, Zhang Wenqi, Wang Biao, Ma Yuzhen, Su Liya, Dai Yanfeng, Liu Gang
Key Laboratory of Medical Cell Biology, Clinical Medicine Research Center, Affiliated Hospital of Inner Mongolia Medical University, Hohhot, Inner Mongolia, China.
College of Life Science, Inner Mongolia University, Hohhot, Inner Mongolia, China.
Front Vet Sci. 2023 Feb 16;10:1093274. doi: 10.3389/fvets.2023.1093274. eCollection 2023.
In the present study, the synergistic protective effect of co-supplementation of glutathione (GSH) with selenium nanoparticles (SeNPs) on the cryopreservation efficiency of bull semen was analyzed.
After collection, the ejaculates of Holstein bulls were subsequently diluted with a Tris extender buffer supplemented with different concentrations of SeNPs (0, 1, 2, and 4 μg/ml), followed by semen equilibration at 4°C and assessment of sperm viability and motility. Subsequently, the ejaculates of Holstein bulls were pooled, split into four equal groups, and diluted with a Tris extender buffer supplemented with basic extender (negative control group, NC group), 2 μg/ml SeNPs (SeNPs group), 4 mM GSH (GSH group), and 4 mM GSH plus 2 μg/ml SeNPs (GSH + SeNPs group). After cryopreservation, motility, viability, mitochondrial activity, plasma membrane integrity, acrosome integrity, concentration of malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT), and ability of frozen-thawed sperm cells to support embryonic development were evaluated.
No side effect of SeNPs concentrations applied in the current study on the motility and viability of equilibrated bull spermatozoa was found. Meanwhile, supplementation of SeNPs significantly promoted the motility and viability of equilibrated bull spermatozoa. Furthermore, the co-supplementation of GSH with SeNPs effectively protected bull spermatozoa from cryoinjury as expressed by promoting semen motility, viability, mitochondrial activity, plasma membrane integrity, and acrosome integrity. Finally, the enhanced antioxidant capacity and embryonic development potential in the frozen-thawed bull spermatozoa cryopreserved by co-supplementation of GSH with SeNPs further confirmed the synergistic protective effect of co-supplementation of GSH with SeNPs on the cryopreservation of bull semen.
在本研究中，分析了谷胱甘肽（GSH）与硒纳米颗粒（SeNPs）联合补充对公牛精液冷冻保存效率的协同保护作用。
采集后，将荷斯坦公牛的射精精液用添加不同浓度SeNPs（0、1、2和4μg/ml）的Tris稀释缓冲液进行稀释，随后在4°C下进行精液平衡，并评估精子活力和运动能力。随后，将荷斯坦公牛的射精精液汇集，分成四个相等的组，并用添加基础稀释液的Tris稀释缓冲液进行稀释（阴性对照组，NC组）、2μg/ml SeNPs（SeNPs组）、4mM GSH（GSH组）以及4mM GSH加2μg/ml SeNPs（GSH + SeNPs组）。冷冻保存后，评估运动能力、活力、线粒体活性、质膜完整性、顶体完整性、丙二醛（MDA）、超氧化物歧化酶（SOD）和过氧化氢酶（CAT）的浓度，以及冻融精子细胞支持胚胎发育的能力。
未发现本研究中应用的SeNPs浓度对平衡后的公牛精子的运动能力和活力有副作用。同时，补充SeNPs显著促进了平衡后的公牛精子的运动能力和活力。此外，GSH与SeNPs联合补充有效地保护公牛精子免受冷冻损伤，表现为促进精液运动能力、活力、线粒体活性、质膜完整性和顶体完整性。最后，GSH与SeNPs联合补充冷冻保存的冻融公牛精子中增强的抗氧化能力和胚胎发育潜力进一步证实了GSH与SeNPs联合补充对公牛精液冷冻保存的协同保护作用。
