Barakat Sarah, Yang Fan, Yelkenci Hayriye Ecem, Kök Kıvanç, Mann Giovanni E, Eroğlu Emrah
Regenerative and Restorative Medicine Research Center (REMER), Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul, 34810, Turkey.
King's British Heart Foundation Centre of Research Excellence, School of Cardiovascular and Metabolic Medicine & Sciences, Faculty of Life Sciences & Medicine, King's College London, 150 Stamford Street, London, SE1 9NH, UK.
Sci Data. 2025 Jun 12;12(1):989. doi: 10.1038/s41597-025-05160-z.
Hyperoxia in standard cell cultures (18 kPa O) imposes cellular oxidative stress, potentially skewing research and drug screening outcomes. Cerebral microvascular endothelial cells (hCMEC/D3) experience no more than 7 kPa O in vivo. In this study, hCMEC/D3 cells were adapted to 5 kPa O for 5 days to optimize an in vitro physiological cell culture model. Using a SYNAPT G2-Si mass spectrometer, we compared the proteomic profiles of cells cultured under 5 kPa versus 18 kPa O. A substantial proteomic shift under hyperoxia highlighted the strong impact of oxygen levels on protein expression. We further investigated the effect of oxygen levels on drug screening using sulforaphane (SFN), an inducer of NRF2-regulated antioxidant defense genes. SFN induced more pronounced changes in proteomic profiles under 18 kPa O compared to 5 kPa, indicating oxygen-dependent cellular drug responses. This dataset offers a valuable resource for analyzing oxygen-sensitive proteomic changes. Comparative studies using different drugs or cell types could further elucidate oxygen-dependent signaling and inform the development of therapies aligned with physiological oxygen levels.
标准细胞培养环境中的高氧（18 kPa O）会引发细胞氧化应激，这可能会影响研究和药物筛选结果。脑微血管内皮细胞（hCMEC/D3）在体内所接触的氧气压力不超过7 kPa O。在本研究中，hCMEC/D3细胞在5 kPa O的环境中培养5天，以优化体外生理细胞培养模型。我们使用SYNAPT G2-Si质谱仪，比较了在5 kPa和18 kPa O环境下培养的细胞的蛋白质组图谱。高氧环境下蛋白质组发生了显著变化，突出了氧水平对蛋白质表达的强烈影响。我们进一步研究了氧水平对使用萝卜硫素（SFN）进行药物筛选的影响，SFN是一种NRF2调节的抗氧化防御基因的诱导剂。与5 kPa相比，SFN在18 kPa O环境下诱导的蛋白质组图谱变化更为明显，表明细胞对药物的反应具有氧依赖性。该数据集为分析氧敏感的蛋白质组变化提供了宝贵资源。使用不同药物或细胞类型的比较研究可以进一步阐明氧依赖性信号传导，并为与生理氧水平相匹配的治疗方法的开发提供参考。
