Department of Anatomy, Faculty of Medical Science, Tarbiat Modares University, Tehran, Iran; Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
Cryobiology. 2019 Oct;90:89-95. doi: 10.1016/j.cryobiol.2019.07.004. Epub 2019 Jul 19.
Despite advances in vitrification techniques for sperm cryopreservation, cryo-damages of sperm caused by generation of reactive oxygen species (ROS) continue to impede implementation of this technique. This study analyses the effects of taurine and hypotaurine as anti-oxidants during vitrification of human sperms. The study was performed in two steps. In the first step, 20 normospermic semen samples were vitrified in the presence of varying concentrations of taurine and hypotaurine, and their effects as anti-oxidant agents on classical sperm parameters, hyaluronan-binding assay (HBA), lipid peroxidation (LPO) and acrosome reaction (AR) were studied. Statistical analyses showed that the sperm parameters in all vitrified groups decreased significantly (P < 0.05) compared to the fresh group. However, HBA and acrosome integrity in vitrified groups containing taurine and 50 mM of hypotaurine were better than in the control group (P < 0.05). The morphology of the vitrified group was good only in the group that contained 50 mM of hypotaurine (P < 0.05). Based on the results from the first step, 50 mM of hypotaurine was considered the ideal anti-oxidant formulation and further tests were carried out on 10 normospermic semen samples with this protecting agent. In addition to the mentioned parameters, the expression of heat shock proteinA2 (HSPA2) was studied in the vitrified group with 50 mM hypotaurine, warmed under two different warming temperatures 37 and 42 °C. 50 mM Hypotaurine was found to equally improve motility, morphology, HBA, and AR after warming at 37 °C and 42 °C (P < 0.05). However, at both warming temperatures, the expression of HSPA2 was reduced in all vitrified groups comparing to the fresh group (P < 0.05). In conclusion, taurine and hypotaurine antioxidants, especially 50 mM hypotaurine, are able to reduce deleterious cryo-injuries on morphology, acrosome and HBA and improve sperm recovery at both warming temperatures (37 and 42 °C). However, they do not have any protective action on expression of HSPA2.
尽管精子玻璃化冷冻技术取得了进展,但由于活性氧(ROS)的产生而导致的精子冷冻损伤仍在阻碍该技术的应用。本研究分析了牛磺酸和次牛磺酸作为抗氧化剂在人类精子玻璃化过程中的作用。该研究分两步进行。在第一步中,20 份正常精子样本在不同浓度的牛磺酸和次牛磺酸存在的情况下进行玻璃化处理,研究其作为抗氧化剂对经典精子参数、透明质酸结合试验(HBA)、脂质过氧化(LPO)和顶体反应(AR)的影响。统计分析表明,与新鲜组相比,所有玻璃化组的精子参数均显著下降(P<0.05)。然而,含有牛磺酸和 50mM 次牛磺酸的玻璃化组的 HBA 和顶体完整性优于对照组(P<0.05)。只有在含有 50mM 次牛磺酸的玻璃化组中,形态学结果良好(P<0.05)。基于第一步的结果,认为 50mM 次牛磺酸是理想的抗氧化剂配方,并在含有该保护剂的 10 份正常精子样本中进一步进行了测试。除了上述参数外,还研究了含有 50mM 次牛磺酸的玻璃化组中热休克蛋白 A2(HSPA2)的表达,该组在 37 和 42°C 两种不同的升温温度下解冻。发现 50mM 次牛磺酸在 37°C 和 42°C 升温时均能提高活力、形态、HBA 和 AR(P<0.05)。然而,在两种升温温度下,与新鲜组相比,所有玻璃化组的 HSPA2 表达均降低(P<0.05)。总之,牛磺酸和次牛磺酸抗氧化剂,特别是 50mM 次牛磺酸,能够减少形态、顶体和 HBA 的冷冻损伤,并在 37°C 和 42°C 两种升温温度下提高精子回收率。然而,它们对 HSPA2 的表达没有任何保护作用。