精子质量与精子细胞氧化平衡的关系

António Patricio , Daniel Filipe Cruz , Joana Vieira Silva , Ana Padrão , Bárbara Regadas Correia , Luís Korrodi-Gregório , Rita Ferreira , Nuno Maia , Saul Almeida , João Lourenço , Vladimiro Silva , Margarida Fardilha
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引用次数: 17

摘要

不孕不育是一种影响全球约15%育龄夫妇的临床疾病。近年来,人们对氧化应激(OS)对精液质量下降的影响进行了探讨。OS对应于机体中存在的氧化剂和抗氧化剂防御之间的不平衡。高水平的活性氧(ROS)会破坏存在于精子细胞中的生物分子,并可能导致膜完整性丧失、DNA断裂,甚至通过细胞凋亡导致死亡。本研究旨在评估人类精液临床参数与评估OS存在的参数之间的相关性。材料和方法本研究从随机抽取的一组献血者中抽取了32份精液样本。根据世界卫生组织的指导方针分析精液的基本参数。采用比色法和免疫印迹法分别测定精子细胞的总抗氧化能力和某些抗氧化蛋白超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶4 (GPx4)的表达。通过凹槽印迹法测定3-硝基酪氨酸和羰基的存在,分析活性氧对精子蛋白氧化的影响。脂质过氧化评价,通过执行硫代巴比妥酸活性物质(TBARS)测定与比色试验。结果SOD与黏度(p = 0.035)、体积(p = 0.004)、羰基含量(p = 0.005)呈负相关。该蛋白也与精子细胞中尾巴缺陷的存在呈正相关(p = 0.044)。GPx4与非进行性运动精子的存在呈负相关(p = 0.012)。TBARS检测结果显示,精子细胞浓度(p = 0.000)和精子总数(p = 0.026)呈负相关,而精液体积与TBARS检测结果呈正相关(p = 0.038)。结论精子质量受精子细胞氧化平衡的影响。所得结果表明,脂质过氧化导致精子浓度降低;抗氧化蛋白保护精子免受蛋白质氧化,有助于增加精子活力和正常精液粘度。因此,评价氧化参数可能是男性不育症诊断和抗氧化治疗随访的有用工具。
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Relation between seminal quality and oxidative balance in sperm cells

Objectives

Infertility is a clinical disorder affecting approximately 15% of reproductive-aged couples worldwide. Recently, the influence of oxidative stress (OS) in decreased semen quality has been discussed. OS corresponds to an imbalance between oxidants and antioxidants defenses, present in the organism. High levels of reactive oxygen species (ROS) damage biomolecules present in sperm cells and may lead to the loss of membrane integrity, DNA fragmentation or even to death by apoptosis. This study aimed to evaluate the correlation between human semen clinic parameters and parameters that assessed the presence of OS.

Material and methods

A total of 32 semen samples, obtained from a randomized group of donors, were included in this study. Basic semen parameters were analyzed according to the WHO's guidelines. The total antioxidant capacity of sperm cells was measured as well as the expression of certain antioxidant proteins, namely superoxide dismutase (SOD) and glutathione peroxidase 4 (GPx4), by colorimetric techniques and immunoblotting, respectively. The effect of ROS in spermatozoa protein oxidation was analyzed by determining the presence of 3-nitrotyrosine and carbonyl groups, by slot blot. Lipid peroxidation was evaluated, by performing the thiobarbituric acid reactive substances (TBARS) assay with colorimetric tests.

Results

The results indicated that SOD was negatively correlated with viscosity (p = 0.035), volume (p = 0.004) and carbonyl groups presence (p = 0.005). This protein also showed a positive correlation with the presence of tail defects in sperm cells (p = 0.044). In turn, GPx4 showed a negative correlation with the presence of non-progressive motile spermatozoa (p = 0.012). TBARS assay revealed a negative correlation with the concentration of sperm cells (p = 0.000) and the total number of spermatozoa (p = 0.026), but, in turn, this assay showed a positive correlation with the volume of semen (p = 0.038).

Conclusion

It was concluded that the seminal quality is affected by the oxidative balance in sperm cells. The obtained results suggest that lipid peroxidation leads to a reduction in sperm concentration; antioxidant proteins protect the spermatozoa against protein oxidation and contribute to an increased sperm motility and normal semen viscosity. Thus, evaluation of oxidative parameters may be a useful tool for male infertility diagnosis and follow-up of antioxidant treatments.

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