Mohammadi F, Varanloo N, Heydari Nasrabadi M, Vatannejad A, Amjadi F S, Javedani Masroor M, Bajelan L, Mehdizadeh M, Aflatoonian R, Zandieh Z
Anatomy Department, School of Medicine, Iran University of Medical Science, Tehran, Iran.
Shahid Akbar Abadi Clinical Research Development Unit (ShACRDU), Iran University of Medical Sciences (IUMS), Tehran, Iran.
Cell Tissue Bank. 2019 Mar;20(1):77-86. doi: 10.1007/s10561-018-9731-0. Epub 2019 Jan 29.
The aim of this study is to evaluate the beneficial effect of Myoinositol (MYO) supplement in freezing media on the post thaw sperm quality. Semen samples from 40 normozoospermic men were divided into two aliquots and frozen with simple or 2 mg/mL MYO supplemented freezing medium. Post thaw process including, computer-assissted sperm analysis was used to analyze sperm motility and morphology. Reactive oxygen species was evaluated by the fluorometry of DCFH-DA, as well as total antioxidant capacity and lipid peroxidation were measured based on colorimetric assay by ELISA reader. Eventually, DNA fragmentation was assessed using TUNEL staining. MYO significantly improved progressive motility and normal morphology in treated samples (p < 0.05). Lipid peroxidation (malondialdehyde level) can be diminished in samples were frozen by MYO supplemented freezing media (p < 0.05). While MYO did not affect the amount of ROS (p > 0.05), it was associated with high values of total antioxidant capacity (p < 0.05). DNA integrity was significantly affected by MYO, as in MYO treated samples, DNA fragmentation was decreased compared to control ones (p < 0.001). The findings support the use of 2 mg/mL myoinositol supplemented freezing media in sperm cryopreservation to increase sperm quality after freezing-thawing procedures.
本研究的目的是评估在冷冻培养基中添加肌醇(MYO)对解冻后精子质量的有益作用。将40名正常精子男性的精液样本分成两份,分别用简单的冷冻培养基或添加了2mg/mL MYO的冷冻培养基进行冷冻。解冻后过程包括使用计算机辅助精子分析来分析精子活力和形态。通过DCFH-DA荧光测定法评估活性氧,同时基于酶标仪比色法测量总抗氧化能力和脂质过氧化。最终,使用TUNEL染色评估DNA片段化。MYO显著改善了处理后样本中的前向运动能力和正常形态(p<0.05)。在添加MYO的冷冻培养基中冷冻的样本中,脂质过氧化(丙二醛水平)可以降低(p<0.05)。虽然MYO不影响活性氧的量(p>0.05),但它与较高的总抗氧化能力值相关(p<0.05)。MYO显著影响DNA完整性,因为在MYO处理的样本中,与对照样本相比,DNA片段化减少(p<0.001)。这些发现支持在精子冷冻保存中使用添加2mg/mL肌醇 的冷冻培养基,以提高冻融程序后的精子质量。