Abbasi Azam, Nouri Houshang, Samadi Firooz
Department of Animal and Poultry Physiology, Faculty of Animal Science, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Golestan, Iran.
Department of Animal Science, Faculty of Agricultural Science and Engineering, University of Tehran, Karaj, Alborz, Iran.
Anim Reprod Sci. 2025 Jan;272:107653. doi: 10.1016/j.anireprosci.2024.107653. Epub 2024 Nov 26.
Cryopreservation is a crucial method for the preservation of genetic materials. Nevertheless, this technique can have adverse effects due to cryo-oxidative damage. The primary objective of this study was to investigate the cryo-protective effects of niosomal nanocarriers loaded with artemisinin (NN-Art) on the functional attributes of equine spermatozoa. The parameters examined included sperm motility and kinematics through computer-assisted sperm analysis (CASA), plasma membrane integrity via the hypoosmotic swelling test, viability using the eosin-nigrosin staining technique, morphology with Hancock's solution, and lipid peroxidation through the thiobarbituric acid reactive substances assay. Additionally, flow cytometry was applied to assess mitochondrial membrane potential, reactive oxygen species (ROS) levels, sperm chromatin structure, and apoptotic-like changes. The CASA indicated a greater enhancement in the total and progressive motility and as well straight line velocity following the addition of NN-Art into the cryo-diluent medium compared with the conventional form of artemisinin and control (P < 0.05). Moreover, the incorporation of cryo-diluent medium with NN-Art demonstrated a greater efficiency in the improvement of viability, proportion of spermatozoa abnormality, plasma membrane functionality, and lipid peroxidation (P < 0.05). The flow cytometry assessment demonstrated a significant enhancement in mitochondrial potential activity, a reduction in the level of ROS, and an increase in the proportion of live spermatozoa along with a decrease in late apoptotic stallion post-thawed spermatozoa treated with both Art and specifically NN-Art. The results of this study suggest that NN-Art exhibit enhanced antioxidant capabilities, which are crucial for improving the functional quality of equine spermatozoa by mitigating cryo-oxidative stress.
冷冻保存是保存遗传物质的关键方法。然而，由于冷冻氧化损伤，该技术可能会产生不利影响。本研究的主要目的是研究负载青蒿素的囊泡纳米载体（NN-Art）对马精子功能特性的冷冻保护作用。检测的参数包括通过计算机辅助精子分析（CASA）评估精子活力和运动学、通过低渗肿胀试验评估质膜完整性、使用伊红-黑色素染色技术评估活力、用汉考克溶液评估形态，以及通过硫代巴比妥酸反应物质测定评估脂质过氧化。此外，还应用流式细胞术评估线粒体膜电位、活性氧（ROS）水平、精子染色质结构和凋亡样变化。CASA结果表明，与传统形式的青蒿素和对照组相比，在冷冻稀释培养基中添加NN-Art后，总活力、渐进性活力以及直线速度均有更大程度的提高（P<0.05）。此外，在冷冻稀释培养基中加入NN-Art在提高活力、精子异常比例、质膜功能和脂质过氧化方面表现出更高的效率（P<0.05）。流式细胞术评估表明，用青蒿素和特别是NN-Art处理的解冻后种马精子的线粒体电位活性显著增强，ROS水平降低，活精子比例增加，晚期凋亡减少。本研究结果表明，NN-Art具有增强的抗氧化能力，这对于通过减轻冷冻氧化应激来提高马精子的功能质量至关重要。
