Department of Animal Physiology and Endocrinology, University of Agriculture in Krakow, Al. Mickiewicza 21, 31-120, Krakow, Poland.
Department of Animal Physiology and Endocrinology, University of Agriculture in Krakow, Al. Mickiewicza 21, 31-120, Krakow, Poland.
Theriogenology. 2024 Jan 15;214:73-80. doi: 10.1016/j.theriogenology.2023.10.008. Epub 2023 Oct 14.
The imbalance between free reactive oxygen species (ROS) generation and removal (e.g., by antioxidative enzymes) leads to the damage of important biomolecules and cells. Earlier studies in hens showed that treatment with tamoxifen (TMX; estrogen receptor inhibitor) modulates oxidative stress and causes the reproductive system regression realized by cell apoptosis. The aim of the present study was, therefore, to examine the expression and immunolocalization of the key enzymatic antioxidants, i.e. catalase (CAT) and superoxide dismutase (SOD), in the chicken oviduct following TMX treatment. Laying hens were treated daily with TMX until a pause in egg-laying occurred and then euthanized on day 8 of the experiment. Quantitative real-time PCR and western blot analyses showed the presence of CAT and SOD transcripts and proteins, respectively, in all oviductal segments, i.e., the infundibulum, magnum, isthmus, shell gland and vagina. In control hens (laying), the mRNA expression of CAT was the highest in the shell gland, lower in the isthmus and the lowest in other oviductal parts, whereas protein expression was the highest in the magnum, lower in the isthmus and the lowest in other segments. The SOD transcript and protein abundances only were lower in the magnum than in other segments. Immunoreactive CAT and SOD products were localized in all layers of the oviductal wall, but the intensity of staining depended on the cell type. TMX treatment affected CAT and SOD expression and the effect of TMX depended on gene, protein, cell type and oviductal part. Generally, CAT expression was elevated, while SOD expression was decreased under TMX treatment. These results point to the importance of CAT and SOD in the maintenance of proper oviduct health and function. Changes in ROS scavenging enzymes after estrogen receptor blockage indicate the significance of estrogen in the regulation of oxidative status in the avian oviduct.
活性氧（ROS）的产生和清除（例如，通过抗氧化酶）之间的失衡会导致重要生物分子和细胞的损伤。早期在母鸡中的研究表明，他莫昔芬（TMX；雌激素受体抑制剂）的治疗会调节氧化应激并导致细胞凋亡实现的生殖系统退化。因此，本研究的目的是检查 TMX 处理后鸡输卵管中关键酶抗氧化剂，即过氧化氢酶（CAT）和超氧化物歧化酶（SOD）的表达和免疫定位。产蛋母鸡每天接受 TMX 治疗，直到产蛋暂停，然后在实验的第 8 天安乐死。定量实时 PCR 和 Western blot 分析显示，CAT 和 SOD 转录本和蛋白质分别存在于所有输卵管段，即漏斗、阔韧带、峡部、壳腺和阴道。在对照母鸡（产卵）中，CAT 的 mRNA 表达在壳腺中最高，峡部中较低，在其他输卵管段中最低，而蛋白质表达在阔韧带中最高，峡部中较低，在其他段中最低。SOD 转录物和蛋白质丰度仅在阔韧带中低于其他段。免疫反应性 CAT 和 SOD 产物定位于输卵管壁的所有层，但染色强度取决于细胞类型。TMX 处理影响 CAT 和 SOD 的表达，而 TMX 的作用取决于基因、蛋白质、细胞类型和输卵管部位。一般来说，CAT 表达升高，而 SOD 表达在 TMX 处理下降低。这些结果表明 CAT 和 SOD 在维持适当的输卵管健康和功能方面的重要性。雌激素受体阻断后 ROS 清除酶的变化表明雌激素在调节禽类输卵管氧化状态方面的重要性。
