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含有肌醇的冷冻稀释液中冻融公牛精子的微观、流式细胞术和氧化参数评估

Evaluation of Microscopic, Flow Cytometric, and Oxidative Parameters of the Frozen-Thawed Bull Sperm in a Freezing Extender Containing Myo-Inositol.

作者信息

Mohammadi Armin, Asadpour Reza, Tayefi-Nasrabadi Hossein, Rahbar Maryam, Joozani Razi-Jafari

机构信息

Graduated from Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran.

Department of Clinical Science and Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran.

出版信息

Biopreserv Biobank. 2022 Apr;20(2):176-184. doi: 10.1089/bio.2021.0046. Epub 2021 Aug 12.

Abstract

This research was conducted to assess the effect of myo-inositol (MYO) in the freezing extender on the semen quality and oxidative stress parameters of frozen-thawed bull sperm. Semen samples were obtained from four bulls ( = 24, six ejaculates per bull), twice a week, and diluted into four equal aliquots in freezing extenders containing different concentrations of MYO (0, 2, 3, and 4 mg/mL). After a freezing/thawing process, velocity parameters, plasma membrane integrity, apoptosis status, malondialdehyde level, and oxidative stress parameters were assessed. Supplementation of freezing extender with 3 mg/mL MYO resulted in higher rapid motility (62.22% ± 2.63%), progressive motility (77.45% ± 2.65%), viability (78% ± 0.91%), plasma membrane integrity (86 ± 0.85), catalase (20.03 ± 0.39 U/mL) activity, and lower significance of lipid peroxidation (3.60 ± 0.15 nmol/dL) than those of the control group ( < 0.05). A significantly lower percentage of normal morphology and intact acrosomes were observed for frozen-thawed semen in the extender supplemented with 4 mg/mL MYO than those of the control group ( < 0.05). Freezing of the sperm in the extender containing 3 mg/mL of MYO leads to a higher percentage of live cells (38.3 ± 2.76). Beat-cross-frequency, amplitude of lateral head displacement, linearity, total antioxidant capacity, total peroxidase activity, early apoptotic status, and superoxide dismutase activities were not affected by MYO levels in the extenders ( > 0.05). The findings of this study suggest that the supplementation of the freezing extender with 3 mg/mL MYO resulted in a higher quality of frozen-thawed bull sperm.

摘要

本研究旨在评估冷冻稀释液中肌醇(MYO)对冻融公牛精子精液质量和氧化应激参数的影响。每周两次从四头公牛(n = 24,每头公牛六次射精)采集精液样本,并在含有不同浓度MYO(0、2、3和4 mg/mL)的冷冻稀释液中稀释成四个等量的等分试样。经过冷冻/解冻过程后,评估精子的速度参数、质膜完整性、凋亡状态、丙二醛水平和氧化应激参数。在冷冻稀释液中添加3 mg/mL MYO可使快速运动率(62.22% ± 2.63%)、渐进性运动率(77.45% ± 2.65%)、活力(78% ± 0.91%)、质膜完整性(86 ± 0.85)、过氧化氢酶(20.03 ± 0.39 U/mL)活性更高,脂质过氧化的显著程度(3.60 ± 0.15 nmol/dL)低于对照组(P < 0.05)。与对照组相比,在添加4 mg/mL MYO的稀释液中冻融精液的正常形态和完整顶体的百分比显著降低(P < 0.05)。在含有3 mg/mL MYO的稀释液中冷冻精子可使活细胞百分比更高(38.3 ± 2.76)。摆动交叉频率、头部侧向位移幅度、线性度、总抗氧化能力、总过氧化物酶活性、早期凋亡状态和超氧化物歧化酶活性不受稀释液中MYO水平的影响(P > 0.05)。本研究结果表明,在冷冻稀释液中添加3 mg/mL MYO可提高冻融公牛精子的质量。

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