Biology of Reproduction and Stem Cell Group, CNC - Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra.
Andrology. 2013 Sep;1(5):707-14. doi: 10.1111/j.2047-2927.2013.00098.x. Epub 2013 Jul 9.
One of the consequences of oxygen metabolism is the production of reactive oxygen species (ROS) which in a situation of imbalance with antioxidants can damage several biomolecules, compromise cell function and even lead to cellular death. The particularities of the sperm cell make it particularly vulnerable to ROS attack compromising its functionality, mirrored in terms of fertility outcome and making the study of the origin of sperm ROS, as well as the alterations they cause very important. In the present work, we used UVB irradiation, an easy experimental approach known as a potent inducer of ROS formation, to better understand the origin of ROS damage without any confounding effects that usually exist in disease models in which ROS are reported to play a role. To address these issues we evaluated sperm mitochondrial ROS production using the Mitosox Red Probe, mitochondrial membrane potential using the JC-1 probe, lipid peroxidation through BODIPY probe and vitality using PI. We observed that UVB irradiation leads to an increase in sperm mitochondrial ROS production and lipid peroxidation that occur previously to an observable mitochondrial dysfunction. We concluded that sperm UVB irradiation appears to be a good and easily manipulated in vitro model system to study mitochondria-induced oxidative stress in spermatozoa and its consequences, which may be relevant in terms of dissecting the action pathways of many other pathologies, drugs and contaminants, including endocrine disruptors.
氧代谢的一个后果是产生活性氧物种 (ROS)，如果 ROS 与抗氧化剂失衡，就会破坏几种生物分子，损害细胞功能，甚至导致细胞死亡。精子细胞的特殊性使其特别容易受到 ROS 的攻击，从而损害其功能，这反映在生育能力的结果上，因此研究精子 ROS 的来源以及它们引起的变化非常重要。在本工作中，我们使用了紫外线 B 照射，这是一种简单的实验方法，被认为是 ROS 形成的有效诱导剂，以更好地理解 ROS 损伤的来源，而没有疾病模型中通常存在的任何混杂效应，在这些模型中，ROS 被认为起作用。为了解决这些问题，我们使用 Mitosox Red Probe 评估了精子线粒体 ROS 的产生，使用 JC-1 探针评估了线粒体膜电位，使用 BODIPY 探针评估了脂质过氧化，使用 PI 评估了活力。我们观察到，紫外线 B 照射会导致精子线粒体 ROS 的产生和脂质过氧化增加，而线粒体功能障碍是在这之前出现的。我们得出结论，精子紫外线 B 照射似乎是一个很好的、易于操作的体外模型系统，可以研究精子线粒体诱导的氧化应激及其后果，这在分析许多其他病理学、药物和污染物（包括内分泌干扰物）的作用途径方面可能具有相关性。
