Departamento de Química e Ciências Ambientais, Instituto de Biociências, Letras e Ciências Exatas, Universidade Estadual Paulista (UNESP), São José do Rio Preto, Brazil.
Departamento de Química e Ciências Ambientais, Instituto de Biociências, Letras e Ciências Exatas, Universidade Estadual Paulista (UNESP), São José do Rio Preto, Brazil.
Toxicol In Vitro. 2019 Apr;56:62-74. doi: 10.1016/j.tiv.2019.01.006. Epub 2019 Jan 14.
This study investigated the effects of incubation period and melatonin treatment on red blood cell (RBC) metabolism in an auto-incubation model of HO-induced oxidative stress. The study was carried out on three healthy adult donors by incubating RBCs in their own plasma at 37 °C, or under the influence of 1 mM HO with and without 100 μM melatonin at different times (0, 1, 3 and 6 h). We assessed incubation period, treatment, as well as any interaction effects between these predictors on erythrocyte osmoregulation, hemolytic rate, oxidative stress markers, and adenylate nucleotide levels. We did not find any relevant effects of both incubation period and treatments on osmotic, antioxidant and adenylate parameters. On the other hand, hemolysis degree and biomolecule oxidation levels in the plasma increased over time, 3-fold and about 25%, respectively, regardless any treatment influence. HO treatment more than doubled protein carbonyl groups, regardless time in plasma, and in a time-depending way in erythrocyte membrane extract, effects that were neutralized by melatonin treatment. Through multivariate analyses, we could expand the understanding of energy and redox metabolisms in the maintenance of cellular integrity and metabolic homeostasis. Another interesting observation was the 65-75% contribution of the oxidative lesion markers on hemolysis. Hence, these findings suggested a new and more intuitive RBC suspension model and reinforced the beneficial use of melatonin in human disorders.
本研究旨在探讨孵育期和褪黑素处理对 HO 诱导的氧化应激自体孵育模型中红细胞（RBC）代谢的影响。该研究在 3 名健康成年供体中进行，将 RBC 在 37°C 下自身血浆中孵育，或在 1mM HO 影响下孵育，并分别添加和不添加 100μM 褪黑素，孵育时间为 0、1、3 和 6 小时。我们评估了孵育期、处理以及这些预测因子之间的任何相互作用对红细胞渗透调节、溶血率、氧化应激标志物和腺嘌呤核苷酸水平的影响。我们没有发现孵育期和处理对渗透、抗氧化和腺苷酸参数有任何相关影响。另一方面，无论任何处理影响如何，血浆中的溶血程度和生物分子氧化水平随时间呈指数增长，分别增加了 3 倍和约 25%。HO 处理使蛋白羰基基团增加了两倍以上，无论在血浆中存在多长时间，在红细胞膜提取物中都呈时间依赖性增加，而褪黑素处理可中和这种增加。通过多元分析，我们可以更深入地了解能量和氧化还原代谢在维持细胞完整性和代谢动态平衡中的作用。另一个有趣的观察结果是氧化损伤标志物对溶血的贡献率为 65-75%。因此，这些发现提出了一种新的、更直观的 RBC 悬浮模型，并强化了褪黑素在人类疾病中的有益应用。
