Dempsey Bianca, Pereira da Silva Beatriz, Cruz Litiele Cezar, Vileigas Danielle, Silva Amanda R M, Pereira da Silva Railmara, Meotti Flavia Carla
Department of Biochemistry, Institute of Chemistry, University of São Paulo, São Paulo, Brazil.
Department of Biochemistry, Institute of Chemistry, University of São Paulo, São Paulo, Brazil.
Redox Biol. 2025 May;82:103625. doi: 10.1016/j.redox.2025.103625. Epub 2025 Apr 1.
This work aims to understand how normouricemic levels of uric acid can induce endothelial dysfunction seeking global proteomic alterations in Human Umbilical Vein cells (HUVEC). It reveals significant alterations in redox-sensitive and antioxidant proteins, chaperones, and proteins associated with cell migration and adhesion in response to uric acid exposure. Monitoring cellular oxidation with the roGFP2-Grx1 probe proved increased oxidation levels induced by uric acid, which can be attenuated by peroxidasin (PXDN) inhibition, suggesting a regulatory role for PXDN in mitigating oxidative stress induced by uric acid. As a consequence of uric acid oxidation and the formation of reactive intermediate, we identified adducts in proteins (+140 kDa) in a novel post-translation modification named uratylation. Increased misfolded protein levels and p62 aggregation were also found, indicating disturbances in cellular proteostasis. Furthermore, uric acid promoted monocyte adhesion and upregulated ICAM and VCAM protein levels, implicating a pro-inflammatory response in endothelial cells. These findings provide critical insights into the molecular mechanisms underlying vascular damage associated with uric acid.
这项工作旨在了解正常尿酸水平如何诱导内皮功能障碍，为此在人脐静脉细胞（HUVEC）中寻找整体蛋白质组学改变。研究揭示了在氧化还原敏感和抗氧化蛋白、分子伴侣以及与细胞迁移和黏附相关的蛋白中，因尿酸暴露而发生的显著改变。用roGFP2-Grx1探针监测细胞氧化过程，结果表明尿酸可诱导氧化水平升高，而过氧化物酶（PXDN）抑制可减轻这种升高，这表明PXDN在减轻尿酸诱导的氧化应激中具有调节作用。由于尿酸氧化及反应性中间体的形成，我们在一种名为尿酸化的新型翻译后修饰中鉴定出蛋白质（+140 kDa）中的加合物。还发现错误折叠蛋白水平增加和p62聚集，这表明细胞蛋白质稳态受到干扰。此外，尿酸促进单核细胞黏附并上调ICAM和VCAM蛋白水平，这表明内皮细胞存在促炎反应。这些发现为与尿酸相关的血管损伤的分子机制提供了关键见解。
