Michael A, Alexopoulos C, Pontiki E, Hadjipavlou-Litina D, Saratsis P, Boscos C
Faculty of Veterinary Medicine, Aristotle University of Thessaloniki, 541 24 Thessaloniki, Greece.
Theriogenology. 2007 Jul 15;68(2):204-12. doi: 10.1016/j.theriogenology.2007.04.053. Epub 2007 May 30.
The objective of this study was to evaluate post-thaw quality of frozen dog semen processed with diluents containing different antioxidants. Ejaculates were collected, pooled and evaluated for concentration, motility, rapid steady forward movement (RSF movement), viability, acrosomal integrity and by the hypo-osmotic swelling test. Also, superoxide production, hydroxyl radicals and total reactive oxygen species (tROS) were determined. The pool was divided in seven aliquots, for control and test conditions, which were processed for cryopreservation. The sperm pellets were diluted to a final concentration of 200x10(6)sperm/ml with TRIS-glucose-egg yolk extender containing one of the following supplements: vitamin C (1.5mM), NAC (N-acetyl-l-cysteine; 1.5mM), taurine (0.6mM), catalase (300U/ml), vitamin E (0.3mM) and B16 [5-(4-dimethylamino-phenyl)-2-phenyl-penta-2,4-dienoic acid; 0.3mM]. Post-thaw semen evaluation showed that mean (+/-S.E.M.) motility was increased (p<0.001) after addition of catalase (49.75+/-3.63 versus 39.00+/-2.90 in controls), whereas more spermatozoa with RSF movement were observed (p<0.001) after the catalase, NAC and vitamin E treatments (31.75+/-3.46, 28.00+/-3.27, 26.75+/-3.15, respectively, versus 17.00+/-2.26 in controls). Viability was increased (p<0.001) after addition of catalase, taurine, NAC and tocopherol (66.00+/-3.03, 61.90+/-2.48, 60.60+/-1.93 and 60.50+/-4.12, respectively, versus 51.70+/-2.81 in controls). The percentage of swollen spermatozoa was increased after addition of catalase and taurine (61.75+/-1.61 and 61.25+/-1.49, respectively, versus 55.65+/-1.64 in controls). Acrosomal integrity was not influenced in any case. B16 addition had adverse effects on all parameters evaluated. None of the reactive oxygen species were significantly reduced post-thaw in antioxidant treated semen. The results suggest that catalase had the most pronounced effect in improving post-thaw quality of canine spermatozoa.
本研究的目的是评估用含有不同抗氧化剂的稀释液处理后的冷冻犬精液解冻后的质量。采集精液，合并后评估其浓度、活力、快速稳定向前运动（RSF运动）、存活率、顶体完整性，并通过低渗肿胀试验进行评估。此外，还测定了超氧化物生成、羟基自由基和总活性氧（tROS）。将合并的精液分成七等份，用于对照和试验条件，进行冷冻保存处理。将精子沉淀用含有以下补充剂之一的TRIS-葡萄糖-蛋黄稀释液稀释至最终浓度为200×10⁶精子/ml：维生素C（1.5mM）、N-乙酰半胱氨酸（NAC；1.5mM）、牛磺酸（0.6mM）、过氧化氢酶（300U/ml）、维生素E（0.3mM）和B16[5-（4-二甲基氨基苯基）-2-苯基-2,4-戊二烯酸；(0.3mM）。解冻后精液评估显示，添加过氧化氢酶后平均（±标准误）活力增加（p<0.001）（49.75±3.63，而对照组为39.00±2.90），而在过氧化氢酶、NAC和维生素E处理后观察到更多具有RSF运动的精子（p<0.001）（分别为31.75±3.46、28.00±3.27、26.75±3.15，而对照组为17.00±2.26）。添加过氧化氢酶、牛磺酸、NAC和生育酚后存活率增加（p<0.001）（分别为66.00±3.03、61.90±2.48、60.60±1.93和60.50±4.12，而对照组为51.70±2.81）。添加过氧化氢酶和牛磺酸后肿胀精子的百分比增加（分别为61.75±1.61和61.25±1.49，而对照组为55.65±1.64）。在任何情况下顶体完整性均未受影响。添加B16对所有评估参数均有不利影响。在抗氧化剂处理的精液解冻后，活性氧均未显著减少。结果表明，过氧化氢酶在改善犬精子解冻后质量方面具有最显著的效果。
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