Abdolsamadi Mona, Mohammadi Fatemeh, Nashtaei Maryam Shabani, Teimouri Maryam, Sardar Reza, Dayani Maliheh, Haghighi Maryam, Ghasemi Samaneh, Vatannejad Akram, Zandieh Zahra
Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Department of Anatomical Sciences, Iran University of Medical Science, Tehran, Iran.
Cell Tissue Bank. 2020 Mar;21(1):99-106. doi: 10.1007/s10561-019-09801-7. Epub 2019 Dec 16.
Sperm cryopreservation is a routine method in andrology and IVF laboratory. However, the sperm quality and its fertilizing capacity have been decreased during this process. The purpose of this experiment was to determine the role of myoinositol as a supplement in amelioration of total and progressive sperm motility, DNA fragmentation, total antioxidant capacity (TAC), reactive oxygen species (ROS), and lipid peroxidation after the freezing-thawing process on patients with oligoasthenoteratozoospermia (OAT) syndrome. Semen samples obtained from 40 patients were divided into two aliquots and freezed with simple and 2 mg/mL myoinositol (MYO) supplemented freezing media. All samples were thawed and assessed after one month. Semen parameters were analyzed in terms of the motility by CASA, the level of total ROS by fluorimetry, TAC and MDA by colorimetric assay and finally DNA fragmentation by TUNEL assay. Our results clearly showed that MYO could improve total (37.46 vs. 12.91, p < 0.001) and progressive motility (21.92 vs. 6.49, p < 0.001) in experimental group compared to control group. A higher TAC level was observed in the MYO treated group in comparison to control group (1.11 vs. 0.91, p = 0.05). While MYO supplementation could not be effective on ROS level, it reduced DNA fragmentation of sperm after freeze-thaw process (p = 0.01). Therefore, MYO could be a good supplement for sperm freezing to reduce the detrimental effects of freezing process especially on DNA integrity, which is an important factor in the success of ART, in OAT suffered patients.
精子冷冻保存是男科和体外受精实验室的常规方法。然而,在此过程中精子质量及其受精能力会下降。本实验的目的是确定肌醇作为一种补充剂在改善少弱畸精子症(OAT)综合征患者冻融过程后精子的总活力和前向运动能力、DNA片段化、总抗氧化能力(TAC)、活性氧(ROS)以及脂质过氧化方面的作用。从40名患者获得的精液样本分成两份,分别用简单冷冻培养基和添加2mg/mL肌醇(MYO)的冷冻培养基进行冷冻保存。所有样本在一个月后解冻并进行评估。通过计算机辅助精子分析(CASA)分析精液参数中的活力,通过荧光法分析总ROS水平,通过比色法分析TAC和丙二醛(MDA),最后通过末端脱氧核苷酸转移酶介导的缺口末端标记法(TUNEL)分析DNA片段化。我们的结果清楚地表明与对照组相比,实验组中MYO可提高总活力(37.46对12.91, p < 0.001)和前向运动能力(21.92对6.49, p < 0.001)。与对照组相比,在MYO处理组中观察到更高的TAC水平(1.11对0.91, p = 0.05)。虽然补充MYO对ROS水平无效,但它可降低冻融过程后精子的DNA片段化(p = 0.01)。因此,对于OAT患者,MYO可能是精子冷冻保存时的一种良好补充剂,可以减少冷冻过程的有害影响,尤其是对DNA完整性方面的影响,而DNA完整性是辅助生殖技术成功的一个重要因素。