Shafiei Mojtaba, Forouzanfar Mohsen, Hosseini Sayyed Morteza, Esfahani Mohammad Hossein Nasr
Department of Biochemistry, Fars Science and Research Branch, Islamic Azad University, Fars, Iran.
Department of Embryology at Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
Theriogenology. 2015 May;83(8):1321-7. doi: 10.1016/j.theriogenology.2015.01.018. Epub 2015 Jan 21.
Manganese(III) meso-tetrakis(N-ethylpyridinium-2-yl)porphyrin chloride (MnTE) is a cell-permeable superoxide dismutase mimetic agent which can convert superoxide to hydrogen peroxide (H2O2). Supplementation of MnTE to a commercial semen extender can protect sperm from superoxide but not H2O2. Therefore, we proposed that addition of catalase (0.0, 200, or 400 IU/mL) in combination with MnTE (0.1 μM) may further improve the cryopreservation efficiency of goat semen in commercially optimized freezing media such as Andromed. Therefore, ejaculates were obtained from three adult bucks twice a week during the breeding season and diluted with Andromed supplemented with or without MnTE and catalase and were frozen in liquid nitrogen. Sperm parameters and reactive oxygen species contents were evaluated 2 hours after dilution (before freezing) and after freezing/thawing. The results revealed that all the treatments significantly (P ≤ 0.05) improved sperm motility, viability, and membrane integrity after freezing and reduced reactive oxygen species content compared with the control group, but maximum improvement was obtained in MnTE + 400 IU/mL catalase. In addition, supplementation with these antioxidants significantly (P ≤ 0.05) increases the cleavage rate after IVF. In conclusion, the results of present study suggest that addition of antioxidant MnTE or catalase to commercial optimized media, such as Andromed, improves total motility, membrane integrity, and viability of goat semen samples after thawing. But the degree of improvement for these parameters significantly (P ≤ 0.05) higher when MnTE and catalase were simultaneously added to the cryopreservation media.
氯化中-四（N-乙基吡啶-2-基）卟啉锰（III）（MnTE）是一种可透过细胞的超氧化物歧化酶模拟剂，它能将超氧化物转化为过氧化氢（H2O2）。在商业精液稀释剂中添加MnTE可保护精子免受超氧化物影响，但不能抵御H2O2。因此，我们提出在商业优化的冷冻培养基（如Andromed）中添加过氧化氢酶（0.0、200或400 IU/mL）并结合MnTE（0.1 μM），可能会进一步提高山羊精液在冷冻保存时的效率。因此，在繁殖季节每周两次从三只成年公羊采集精液，用添加或不添加MnTE和过氧化氢酶的Andromed进行稀释，然后在液氮中冷冻。在稀释后2小时（冷冻前）以及冷冻/解冻后评估精子参数和活性氧含量。结果显示，与对照组相比，所有处理均显著（P≤0.05）提高了冷冻后精子的活力、存活率和膜完整性，并降低了活性氧含量，但在MnTE + 400 IU/mL过氧化氢酶组中获得了最大改善。此外，添加这些抗氧化剂显著（P≤0.05）提高了体外受精后的卵裂率。总之，本研究结果表明，在商业优化培养基（如Andromed）中添加抗氧化剂MnTE或过氧化氢酶，可提高解冻后山羊精液样本的总活力、膜完整性和存活率。但是，当MnTE和过氧化氢酶同时添加到冷冻保存培养基中时，这些参数的改善程度显著（P≤0.05）更高。
