Rheumatology and Immune-mediated Diseases Group, Galicia Sur Health Research Institute (IIS Galicia Sur), Vigo, Spain.
Rheumatology Department, University Hospital of Vigo, Vigo, Spain.
Front Immunol. 2023 Dec 14;14:1277267. doi: 10.3389/fimmu.2023.1277267. eCollection 2023.
Endothelial cell (EC) dysfunction is a hallmark of Systemic Lupus Erythematosus (SLE) and Tie2 is a receptor essential for vascular stability. Inflammatory processes promote inhibition of Tie2 homeostatic activation, driving vascular dysfunction. In this work we determined whether type I Interferons (IFN) induce Tie2 signalling-mediated endothelial dysfunction in patients with SLE. Serum levels of Angiopoietin (Ang)-1, Ang-2 and soluble (s)Tie1 in patients with SLE and healthy controls were measured by ELISA. Monocytes from patients with SLE and Human Umbilical Vein EC (HUVEC) were stimulated with IFN-α, IFN-β (1000 I.U.) or SLE serum (20%). mRNA and protein expression, phosphorylation and translocation were determined by quantitative PCR, ELISA, Western Blot, flow cytometry and confocal microscopy. Viability and angiogenic capacity were determined by calcein and tube formation assays. We found that sTie1 and Ang-2 serum levels were increased and Ang-1 decreased in patients with SLE and were associated with clinical characteristics. Type I IFN significantly decreased Ang-1 and increased Ang-2 in monocytes from patients with SLE. Type I IFN increased sTie1 and Ang-2 secretion and reduced Tie2 activation in HUVEC. Functionally, type I IFN significantly reduced EC viability and impaired angiogenesis in a Tie2 signalling-dependent manner. Finally, SLE serum increased Ang-2 and sTie1 secretion and significantly decreased tube formation. Importantly, Tie1 and IFNAR1 knockdown reversed these effects in tube formation. Overall, type I IFN play an important role in the stability of EC by inhibiting Tie2 signalling, suggesting that these processes may be implicated in the cardiovascular events observed in patients with SLE.
内皮细胞 (EC) 功能障碍是系统性红斑狼疮 (SLE) 的标志,而 Tie2 是血管稳定性所必需的受体。炎症过程促进 Tie2 稳态激活的抑制,导致血管功能障碍。在这项工作中,我们确定了 I 型干扰素 (IFN) 是否会诱导 SLE 患者的 Tie2 信号介导的内皮功能障碍。通过 ELISA 测定 SLE 患者和健康对照者血清中血管生成素 (Ang)-1、Ang-2 和可溶性 (s)Tie1 的水平。用 IFN-α、IFN-β (1000 I.U.) 或 SLE 血清 (20%) 刺激 SLE 患者的单核细胞和人脐静脉内皮细胞 (HUVEC)。通过定量 PCR、ELISA、Western blot、流式细胞术和共聚焦显微镜测定 mRNA 和蛋白表达、磷酸化和转位。通过钙黄绿素和管形成测定法测定细胞活力和血管生成能力。我们发现,SLE 患者的 sTie1 和 Ang-2 血清水平升高,Ang-1 降低,且与临床特征相关。I 型 IFN 显著降低了 SLE 患者单核细胞中的 Ang-1 并增加了 Ang-2。I 型 IFN 增加了 HUVEC 中 sTie1 和 Ang-2 的分泌,并降低了 Tie2 的激活。功能上,I 型 IFN 显著降低了 EC 的活力,并以 Tie2 信号依赖性方式损害了血管生成。最后,SLE 血清增加了 Ang-2 和 sTie1 的分泌,并显著减少了管形成。重要的是,Tie1 和 IFNAR1 的敲低逆转了管形成中的这些作用。总之,I 型 IFN 通过抑制 Tie2 信号在 EC 的稳定性中发挥重要作用,这表明这些过程可能与 SLE 患者观察到的心血管事件有关。