Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, University of Murcia, Campus de Espinardo, E-30100 Murcia, Spain.
Reproduction. 2010 Mar;139(3):557-64. doi: 10.1530/REP-09-0274. Epub 2009 Dec 23.
This study evaluated whether pentoxifylline added to freezing and thawing extenders influenced the function of boar spermatozoa. In Experiment 1, pooled ejaculated sperm-rich fractions were frozen in 0.5 ml straws after dilution in extender supplemented with pentoxifylline to a final concentration of 0, 2, 4, 8, 16 or 32 mM. The addition of 4, 8, 16 and 32 mM pentoxifylline to the freezing extender significantly decreased the progressive and total motility of spermatozoa. The percentage of viable spermatozoa with intact acrosomes as well as the penetration rate and the efficiency of fertilisation were significantly lower in pentoxifylline-treated groups compared with the untreated control. In Experiment 2, a pool of three straws with 'good' post-thaw sperm quality parameters and another three straws with 'poor' sperm quality were diluted in extender with 0, 1, 2, 4, 8, 16 or 32 mM pentoxifylline. Post-thaw samples with both 'good' and 'poor' sperm quality with 0, 2, 4, 8 and 16 mM were used to assess IVF parameters. The addition of pentoxifylline to post-thaw extender did not improve the post-thaw motility or viability of spermatozoa compared with the control. The in vitro penetration was higher (P<0.05) than the control for oocytes fertilised with spermatozoa that were thawed and incubated in extender with 4, 8 and 16 mM pentoxifylline. However, no differences were observed in the efficiency of fertilisation. We conclude that pentoxifylline, as a supplement added to the freezing extender, has a deleterious effect and that it does not improve the survival or in vitro fertilising efficiency of frozen-thawed boar spermatozoa when added after thawing.
本研究评估了在冷冻和解冻液中添加己酮可可碱是否会影响猪精子的功能。在实验 1 中,将混合的富含精子的射出精液部分在添加终浓度为 0、2、4、8、16 或 32mM 己酮可可碱的稀释液中于 0.5ml 细管中冷冻。在冷冻液中添加 4、8、16 和 32mM 己酮可可碱显著降低了精子的前向运动和总运动能力。与未经处理的对照组相比,经己酮可可碱处理的精子的活率、顶体完整率、穿透率和受精效率均显著降低。在实验 2 中,将三支“良好”解冻后精子质量参数的细管和三支“较差”精子质量参数的细管的精液混合,用含有 0、1、2、4、8、16 或 32mM 己酮可可碱的稀释液进行稀释。用含有 0、2、4、8 和 16mM 己酮可可碱的解冻后具有“良好”和“较差”精子质量的样品进行 IVF 参数评估。与对照组相比,解冻液中添加己酮可可碱并没有改善解冻后精子的运动能力或活力。在体外穿透率方面,与对照组相比,用含有 4、8 和 16mM 己酮可可碱的稀释液解冻和孵育的精子对卵母细胞的穿透率更高(P<0.05)。但是,受精效率没有差异。我们的结论是,作为冷冻液添加剂的己酮可可碱具有有害作用,并且在解冻后添加时,不会提高冷冻解冻猪精子的存活或体外受精效率。