Research Unit, Fundación para la Investigación Biomédica del Hospital Universitario Príncipe de Asturias, Alcalá de Henares, Madrid, Spain.
Nephrology Section, Fundación para la Investigación Biomédica del Hospital Universitario Príncipe de Asturias, Alcalá de Henares, Madrid, Spain.
Cardiovasc Res. 2017 Feb;113(2):207-221. doi: 10.1093/cvr/cvw239. Epub 2016 Dec 25.
To analyse the ability of TWEAK to modify the endothelin system, particularly endothelin-1 (ET-1) and endothelin-converting enzyme-1 (ECE-1), studying the intracellular mechanisms implied. TNF-like weak inducer of apoptosis (TWEAK) is a member of TNF superfamily; it has different biological functions such as inflammation, angiogenesis, proliferation, and apoptosis. TWEAK and fibroblast growth-factor-inducible 14 are expressed in different cell types, including endothelial and smooth muscle cells. Despite their presence in endothelial cells, the effect of TWEAK on endothelial function is incompletely defined.
In cells, TWEAK induced protein (Western blot) and mRNA (quantitative polymerase chain reaction) expression of ECE-1. Results were related to transcriptional changes, as ECE-1 promoter activity (transfection assays) was also increased. Transfections with serial deletions of ECE-1 promoter suggest a potential role for AP-1 and NFkB, which were confirmed by electrophoretic mobility shift assays. When AP-1 or NFkB activations were inhibited by specific inhibitors of AP-1, PD-98059 (Erk1/2 inhibitor), or SP-600125 (JNK inhibitor), and also with an inhibitor of NFKB and PDTC, TWEAK effect was partially blocked in both cases, suggesting that both transcription factors are implied in ECE-1 regulation. Moreover, the endothelial changes induced by TWEAK were also tested in vivo, using 3-month-old male CD-1 mice treated with TWEAK 10 µg/kg body weight for 24 h, finding similar effects, a rise in ET-1 production (enzyme-linked immunosorbent assay), and ECE-1 expression in aorta and lung tissues. Mice showed slight hypertension after 4 h of treatment, which disappeared at 24 h.
In pathological situations such as chronic inflammation, TWEAK could be more harmful through this effect at endothelial level. Pharmacological blockade of this cytokine could prevent the haemodynamic and structural changes related to an increased ET-1 synthesis.
分析 TWEAK 修饰内皮素系统的能力，特别是内皮素-1（ET-1）和内皮素转换酶-1（ECE-1），研究所涉及的细胞内机制。TNF 样凋亡弱诱导剂（TWEAK）是 TNF 超家族的一员；它具有多种生物学功能，如炎症、血管生成、增殖和凋亡。TWEAK 和成纤维细胞生长因子诱导 14 在包括内皮细胞和平滑肌细胞在内的不同细胞类型中表达。尽管它们存在于内皮细胞中，但 TWEAK 对内皮功能的影响尚未完全确定。
在细胞中，TWEAK 诱导 ECE-1 的蛋白（Western blot）和 mRNA（定量聚合酶链反应）表达。结果与转录变化有关，因为 ECE-1 启动子活性（转染试验）也增加了。ECE-1 启动子的串联缺失转染表明 AP-1 和 NFkB 可能发挥作用，电泳迁移率变动分析证实了这一点。当用 AP-1 或 NFkB 的特定抑制剂 PD-98059（Erk1/2 抑制剂）或 SP-600125（JNK 抑制剂）抑制 AP-1 或 NFkB 的激活时，TWEAK 的作用在两种情况下都被部分阻断，这表明这两种转录因子都与 ECE-1 的调节有关。此外，还在体内使用 3 个月大的雄性 CD-1 小鼠进行了 TWEAK 10μg/kg 体重治疗 24 小时的实验，以测试 TWEAK 引起的内皮变化，发现 ET-1 产生（酶联免疫吸附试验）增加，以及主动脉和肺组织中 ECE-1 表达增加。小鼠在治疗后 4 小时出现轻度高血压，24 小时后消失。
在慢性炎症等病理情况下，TWEAK 通过这种内皮水平的作用可能会更具危害性。这种细胞因子的药理学阻断可能会阻止与 ET-1 合成增加相关的血流动力学和结构变化。
