衰老和与生活方式有关的疾病,比如心血管疾病,糖尿病,癌症,和神经退行性疾病,是全球面临的重大健康挑战。这些情况通常与氧化还原失衡有关,细胞无法调节反应性氧化还原物质(RRS),导致氧化应激和细胞损伤。尽管已知抗氧化剂可以中和有害的RRS,其临床疗效仍不一致.这种不一致的一个原因是当前体外模型不足以准确模拟体内氧化还原条件。这项研究通过使用代谢引发的人真皮成纤维细胞(NHDF)解决了理解细胞中氧化还原反应异质性的差距,与精确线粒体医学相关的模型。我们调查了代谢启动,增强线粒体生物能学,影响氧化还原对过氧化氢(H2O2)和叔丁基过氧化氢(tBHP)诱导的氧化应激的反应。具体来说,我们探讨了细胞群密度和细胞周期分布对氧化还原动力学的影响。我们的发现表明,在氧化磷酸化促进培养基(OXm)中培养的NHDF细胞在氧化应激反应中表现出明显更大的变异性。这种可变性表明,增强的线粒体生物能量需要不断调节细胞氧化还原机制,可能导致异质反应。此外,在OXm中生长的细胞显示线粒体极化增加,G2/M期细胞百分比较低,导致观察到的异质性。影响这种变异性的关键因素包括氧化剂暴露时的细胞群密度和细胞周期分布的波动。我们的结果强调了在代谢引发模型中使用多种氧化剂以全面了解氧化应激反应和氧化还原调节机制的必要性。此外,该研究强调需要完善体外模型,以更好地反映体内条件,这对于开发有效的基于氧化还原的治疗策略至关重要。
Aging and lifestyle-related diseases, such as cardiovascular diseases, diabetes, cancer, and neurodegenerative disorders, are major global health challenges. These conditions are often linked to
redox imbalances, where cells fail to regulate reactive
redox species (RRS), leading to oxidative stress and cellular damage. Although antioxidants are known to neutralize harmful RRS, their clinical efficacy remains inconsistent. One reason for this inconsistency is the inadequacy of current in vitro models to accurately mimic in vivo
redox conditions. This study addresses the gap in understanding the heterogeneity of
redox responses in cells by using metabolically primed human dermal fibroblasts (NHDF), a model relevant for precision mitochondrial medicine. We investigated how metabolic priming, which enhances mitochondrial bioenergetics, influences redox responses to oxidative stress induced by hydrogen peroxide (H2O2) and tert-butyl hydroperoxide (tBHP). Specifically, we explored the impact of cell population density and cell cycle distribution on
redox dynamics. Our findings indicate that NHDF cells cultured in oxidative phosphorylation-promoting medium (OXm) exhibit significantly larger variability in oxidative stress responses. This variability suggests that enhanced mitochondrial bioenergetics necessitates a constant regulation of the cellular redox machinery, potentially leading to heterogeneous responses. Additionally, cells grown in OXm showed increased mitochondrial polarization and a lower percentage of cells in the G2/M phase, contributing to the observed heterogeneity. Key factors influencing this variability included cell population density at the time of oxidant exposure and fluctuations in cell cycle distribution. Our results highlight the necessity of employing multiple oxidants in metabolic priming models to achieve a comprehensive understanding of oxidative stress responses and
redox regulation mechanisms. Furthermore, the study emphasizes the need to refine in vitro models to better reflect in vivo conditions, which is crucial for the development of effective redox-based therapeutic strategies.