Hematopathology, Centre Diagnòstic Biomèdic (CDB), Hospital Clinic, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Universitat de Barcelona (UB), Barcelona, Spain.
Josep Carreras Leukaemia Research Institute, Hospital Clinic/University of Barcelona Campus, Barcelona, Spain.
J Cell Mol Med. 2020 Jan;24(2):1713-1723. doi: 10.1111/jcmm.14865. Epub 2019 Nov 28.
Endothelial dysfunction is an earlier contributor to the development of atherosclerosis in chronic kidney disease (CKD), in which the role of epigenetic triggers cannot be ruled out. Endothelial protective strategies, such as defibrotide (DF), may be useful in this scenario. We evaluated changes induced by CKD on endothelial cell proteome and explored the effect of DF and the mechanisms involved. Human umbilical cord vein endothelial cells were exposed to sera from healthy donors (n = 20) and patients with end-stage renal disease on haemodialysis (n = 20). Differential protein expression was investigated by using a proteomic approach, Western blot and immunofluorescence. HDAC1 and HDAC2 overexpression was detected. Increased HDAC1 expression occurred at both cytoplasm and nucleus. These effects were dose-dependently inhibited by DF. Both the HDACs inhibitor trichostatin A and DF prevented the up-regulation of the endothelial dysfunction markers induced by the uraemic milieu: intercellular adhesion molecule-1, surface Toll-like receptor-4, von Willebrand Factor and reactive oxygen species. Moreover, DF down-regulated HDACs expression through the PI3/AKT signalling pathway. HDACs appear as key modulators of the CKD-induced endothelial dysfunction as specific blockade by trichostatin A or by DF prevents endothelial dysfunction responses to the CKD insult. Moreover, DF exerts its endothelial protective effect by inhibiting HDAC up-regulation likely through PI3K/AKT.
内皮功能障碍是慢性肾脏病 (CKD) 中动脉粥样硬化发展的早期因素，其中表观遗传触发因素的作用不容忽视。内皮保护策略，如去纤维蛋白（DF），在这种情况下可能是有用的。我们评估了 CKD 对内皮细胞蛋白质组的影响，并探讨了 DF 的作用及其涉及的机制。将人脐静脉内皮细胞暴露于健康供体（n=20）和血液透析终末期肾病患者（n=20）的血清中。通过蛋白质组学方法、Western blot 和免疫荧光检测差异蛋白表达。检测到 HDAC1 和 HDAC2 的过表达。HDAC1 的表达在细胞质和细胞核中均增加。DF 可呈剂量依赖性抑制这些作用。HDAC 抑制剂曲古抑菌素 A 和 DF 均可预防尿毒症环境诱导的内皮功能障碍标志物的上调：细胞间黏附分子-1、表面 Toll 样受体-4、血管性血友病因子和活性氧。此外，DF 通过 PI3/AKT 信号通路下调 HDACs 的表达。HDACs 似乎是 CKD 诱导的内皮功能障碍的关键调节剂，因为曲古抑菌素 A 或 DF 的特异性阻断可防止内皮功能障碍对 CKD 损伤的反应。此外，DF 通过抑制 HDAC 的上调发挥其内皮保护作用，可能通过 PI3K/AKT。
