Nasiri-Foomani Niloofar, Hassani Saeed, Najafi Mojtaba, Samadi Firooz
Department of Animal and Poultry Physiology, Faculty of Animal Science, Gorgan University of Agricultural Science and Natural Resources, Golestan, Gorgan, Iran.
Department of Animal and Poultry Physiology, Faculty of Animal Science, Gorgan University of Agricultural Science and Natural Resources, Golestan, Gorgan, Iran.
Cryobiology. 2025 Mar;118:105188. doi: 10.1016/j.cryobiol.2024.105188. Epub 2024 Dec 29.
The optimization of cryopreservation media to reduce oxidative damage on post-thaw spermatozoa is crucial. This research aimed to assess the antioxidant properties of curcumin-loaded niosomal nanocarriers (CurLNN) on the functional characteristics, the relative expression of apoptotic genes, and flow cytometry assessments of apoptotic-like changes, reactive oxygen species production (ROS), mitochondrial membrane potential, and chromatin integrity in stallion spermatozoa following thawing. Twenty-five ejaculates were diluted in INRA96 freezing media supplemented with 20 μM of either curcumin (Cur) or CurLNN and then cryopreserved. Results demonstrated that spermatozoa treated with Cur, particularly CurLNN, exhibited higher percentages of total and progressive motility, as well as VAP, VSL, and STR kinematics. Additionally, the functionality of the plasma membrane was enhanced, and there was a decrease in spermatozoa abnormality (P < 0.05). The incorporation of cryo-diluent medium with Cur and CurLNN led to increased viability (P < 0.05), while simultaneously reducing the levels of MDA. Flow cytometry analysis revealed a significant enhancement in mitochondrial potential activity, a reduction (P < 0.05) in ROS production, and an increase (P < 0.05) in the proportion of live and a decrease in late apoptotic stallion post-thawed spermatozoa treated with both Cur and CurLNN. Moreover, the relative expression of the Bcl anti-apoptotic gene increased (P < 0.05) by the addition of cur and CurLNN in cryo-diluent extender, while inclusion of CurLNN in cryo-diluent medium resulted in a significant reduction (P < 0.05) in the relative expression of the Bax pro-apoptotic gene in stallion post-thawed spermatozoa. In summary, the findings of this study demonstrated that CurLNN exhibits enhanced antioxidant properties, which contribute to the improved functional quality of spermatozoa by alleviating oxidative stress during the cryopreservation process.
优化冷冻保存介质以减少解冻后精子的氧化损伤至关重要。本研究旨在评估负载姜黄素的脂质体纳米载体（CurLNN）对解冻后种马精子的功能特性、凋亡基因的相对表达、凋亡样变化的流式细胞术评估、活性氧产生（ROS）、线粒体膜电位和染色质完整性的抗氧化特性。将25份射精样本在补充有20μM姜黄素（Cur）或CurLNN的INRA96冷冻培养基中稀释，然后进行冷冻保存。结果表明，用Cur特别是CurLNN处理的精子表现出更高的总活力和渐进性活力百分比，以及VAP、VSL和STR运动学参数。此外，质膜的功能得到增强，精子异常率降低（P<0.05）。在冷冻稀释培养基中加入Cur和CurLNN可提高活力（P<0.05），同时降低丙二醛水平。流式细胞术分析显示，用Cur和CurLNN处理的解冻后种马精子的线粒体电位活性显著增强，ROS产生减少（P<0.05），活细胞比例增加（P<0.05），晚期凋亡细胞减少。此外，在冷冻稀释剂中添加Cur和CurLNN可使Bcl抗凋亡基因的相对表达增加（P<0.05），而在冷冻稀释培养基中加入CurLNN可使解冻后种马精子中促凋亡基因Bax的相对表达显著降低（P<0.05）。总之，本研究结果表明，CurLNN具有增强的抗氧化特性，通过减轻冷冻保存过程中的氧化应激，有助于提高精子的功能质量。
