Figuer Andrea, Santos Fátima M, Ciordia Sergio, Valera Gemma, Martín-Jouve Beatriz, Hernández-Fonseca Juan Pablo, Bodega Guillermo, Ceprián Noemí, Ramírez Rafael, Carracedo Julia, Alique Matilde
Departamento de Biología de Sistemas, Universidad de Alcalá, 28871 Alcala de Henares, Madrid, Spain; Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS), 28034 Madrid, Spain.
Cellular Biology in Renal Diseases Laboratory, IIS-Fundación Jiménez Díaz-Universidad Autónoma Madrid, 28040 Madrid, Spain; Functional Proteomics Laboratory, Centro Nacional de Biotecnología, CSIC, Calle Darwin 3, Campus de Cantoblanco, 28049 Madrid, Spain.
Life Sci. 2024 Aug 15;351:122810. doi: 10.1016/j.lfs.2024.122810. Epub 2024 Jun 11.
Cardiovascular pathology is the main cause of death in chronic kidney disease (CKD) patients. CKD is associated with the accumulation of uremic toxins in the bloodstream, and indoxyl sulfate (IS) is one of the most abundant uremic toxins found in the blood of CKD patients. We conducted an in vitro study to assess the mechanisms underlying the IS-induced endothelial dysfunction that could lead to cardiovascular diseases. We also studied their extracellular vesicles (EVs) owing to their capacity to act as messengers that transmit signals through their cargo.
EVs were characterized by nanoparticle tracking analysis, transmission electron microscopy, flow cytometry, and tetraspanin expression. Cell lysates and isolated EVs were analyzed using liquid chromatography coupled with mass spectrometry, followed by Gene Set Enrichment Analysis to identify the altered pathways.
Proteomic analysis of endothelial cells revealed that IS causes an increase in proteins related to adipogenesis, inflammation, and xenobiotic metabolism and a decrease in proliferation. Extracellular matrix elements, as well as proteins associated with myogenesis, response to UV irradiation, and inflammation, were found to be downregulated in IS-treated EVs. Fatty acid metabolism was also found to be increased along with adipogenesis and inflammation observed in cells.
The treatment of endothelial cells with IS increased the expression of proteins related to adipogenesis, inflammation, and xenobiotic metabolism and was less associated with proliferation. Furthermore, EVs from cells treated with IS may mediate endothelial dysfunction, since they present fewer extracellular matrix elements, myogenesis, inflammatory factors, and proteins downregulated in response to UV radiation.
心血管病理是慢性肾脏病（CKD）患者的主要死亡原因。CKD与血液中尿毒症毒素的蓄积有关，而硫酸吲哚酚（IS）是在CKD患者血液中发现的含量最为丰富的尿毒症毒素之一。我们进行了一项体外研究，以评估IS诱导的可能导致心血管疾病的内皮功能障碍的潜在机制。我们还对其细胞外囊泡（EVs）进行了研究，因为它们有能力作为通过其携带物质传递信号的信使。
通过纳米颗粒跟踪分析、透射电子显微镜、流式细胞术和四跨膜蛋白表达对EVs进行表征。使用液相色谱-质谱联用分析细胞裂解物和分离的EVs，随后进行基因集富集分析以确定改变的途径。
内皮细胞的蛋白质组学分析表明，IS导致与脂肪生成、炎症和外源性物质代谢相关的蛋白质增加，而增殖相关蛋白质减少。在经IS处理的EVs中，细胞外基质成分以及与肌生成、紫外线照射反应和炎症相关的蛋白质被发现下调。脂肪酸代谢也随着细胞中观察到的脂肪生成和炎症增加而增加。
用IS处理内皮细胞会增加与脂肪生成、炎症和外源性物质代谢相关的蛋白质表达，且与增殖的相关性较小。此外，来自经IS处理的细胞的EVs可能介导内皮功能障碍，因为它们呈现较少的细胞外基质成分、肌生成、炎症因子以及对紫外线辐射下调的蛋白质。
