Shalaby Fatma Mohsen, Hassan Soha A, El-Raghi Ali A, Alajmi Fatemah Enad, Alfayoumi Mohamed G, Attia Kandil Abd El Hai
Department of Zoology, Faculty of Sciences, Mansoura University, Dakahlia, Egypt.
Department of Biology, Faculty of Sciences, King Khalid University, Abha, Saudi Arabia.
Anim Biosci. 2025 Sep;38(9):1894-1905. doi: 10.5713/ab.25.0067. Epub 2025 Apr 11.
This study was conducted to investigate the effect of pomegranate oil nanoemulsion (PO-NE) supplementation in semen extender on semen quality, redox status and apoptotic genes of cryopreserved bovine semen.
Seven proven fertility Holstein Friesian bulls (4 to 6 years) were involved, and the semen samples were collected using the artificial vagina method. Semen was pooled and cryopreserved in tris extender containing PO-NE at 0 (PO-NE0), 1 (PO-NE1), 2 (PO-NE 2) and 4 μg/mL (PO-NE4), respectively.
Incorporating 2 or 4 μg/mL PO-NE into freezing media significantly increased sperm progressive motility, viability, membrane integrity, and kinematic parameters. Furthermore, the aforementioned two treated concentrations demonstrated superior antioxidative activities (total antioxidant capacity and super oxide dismutase) and higher nitric oxide levels compared to the control group (p<0.05). The levels of hydrogen peroxide, malondialdehyde, and Nuclear Factor-Kappa B were notably lower in the PO-NE4 treated group compared to the control group. The addition of 2 or 4 μg/mL of PO-NE to the freezing media significantly downregulated pro-apoptotic genes (caspase 3, Bax), while significantly induced the expression of anti-apoptotic gene Bcl2. The addition of PO-NE preserved plasma membrane and acrosome integrity and maintained the ultrastructure of sperm, contrasting with PO-NE0, which exhibited the most damage.
Supplementing the bovine freezing extender with 2 or 4 μg/mL of PO-NE enhanced post-thawed sperm characteristics by reducing oxidative stress, improving antioxidant indices and apoptotic genes expression, and preserving the ultrastructure integrity of frozen-thawed cattle sperm.
本研究旨在探讨在精液稀释液中添加石榴籽油纳米乳剂（PO-NE）对冷冻保存的牛精液质量、氧化还原状态和凋亡基因的影响。
选用7头经证实具有生育能力的荷斯坦弗里生公牛（4至6岁），采用人工阴道法采集精液样本。精液混合后分别在含有0（PO-NE0）、1（PO-NE1）、2（PO-NE2）和4μg/mL（PO-NE4）PO-NE的Tris稀释液中冷冻保存。
在冷冻培养基中加入2或4μg/mL PO-NE可显著提高精子的前向运动能力、活力、膜完整性和运动学参数。此外，与对照组相比，上述两种处理浓度表现出更强的抗氧化活性（总抗氧化能力和超氧化物歧化酶）和更高的一氧化氮水平（p<0.05）。与对照组相比，PO-NE4处理组的过氧化氢、丙二醛和核因子-κB水平显著降低。在冷冻培养基中添加2或4μg/mL PO-NE可显著下调促凋亡基因（半胱天冬酶3、Bax）的表达，同时显著诱导抗凋亡基因Bcl2的表达。添加PO-NE可保持质膜和顶体完整性，并维持精子的超微结构，而PO-NE0组损伤最为严重。
在牛冷冻稀释液中添加2或4μg/mL PO-NE可通过降低氧化应激、改善抗氧化指标和凋亡基因表达以及保持冻融牛精子的超微结构完整性来提高解冻后精子的特性。
