Center for Translational Medicine, Thomas Jefferson University, Philadelphia, PA (G.Z., Q.Q., C.Z., X.S., K.K., B.Y., F.C., Z.-F.G., J.S.).
The Institute of Cardiothoracic Surgery, Changhai Hospital, Naval Medical University, Shanghai, China (G.Z.).
Circ Res. 2023 Feb 3;132(3):306-319. doi: 10.1161/CIRCRESAHA.122.321837. Epub 2022 Dec 23.
NDRG-1 (N-myc downstream-regulated gene 1) is a member of NDRG family that plays essential roles in cell differentiation, proliferation, and stress responses. Although the expression of NDRG1 is regulated by fluid shear stress, its roles in vascular biology remain poorly understood. The purpose of the study is to determine the functional significance of NDRG1 in vascular inflammation and remodeling.
By using quantitative polymerase chain reaction, western blot, and immunohistochemistry, we demonstrate that the expression of NDRG1 is markedly increased in cytokine-stimulated endothelial cells and in human and mouse atherosclerotic lesions. To determine the role of NDRG1 in endothelial activation, we performed loss-of-function studies using NDRG1 short hairpin RNA. Our results demonstrate that NDRG1 knockdown by lentivirus bearing NDRG1 short hairpin RNA substantially attenuates both IL-1β (interleukin-1β) and TNF-α (tumor necrosis factor-α)-induced expression of cytokines/chemokines and adhesion molecules. Intriguingly, inhibition of NDRG1 also significantly attenuates the expression of procoagulant molecules, such as PAI-1 (plasminogen activator inhibitor type 1) and TF (tissue factor), and increases the expression of TM (thrombomodulin) and t-PA (tissue-type plasminogen activator), thus exerting potent antithrombotic effects in endothelial cells. Mechanistically, we showed that NDRG1 interacts with orphan Nur77 (nuclear receptor) and functionally inhibits the transcriptional activity of Nur77 and NF-κB (nuclear factor Kappa B) in endothelial cells. Moreover, in NDRG1 knockdown cells, both cytokine-induced mitogen-activated protein kinase activation, c-Jun phosphorylation, and AP-1 (activator protein 1) transcriptional activity are substantially inhibited. Neointima and atherosclerosis formation induced by carotid artery ligation and arterial thrombosis were markedly attenuated in endothelial cell-specific NDRG1 knockout mice compared with their wild-type littermates.
Our results for the first time identify NDRG1 as a critical mediator implicated in regulating endothelial inflammation, thrombotic responses, and vascular remodeling, and suggest that inhibition of NDRG1 may represent a novel therapeutic strategy for inflammatory vascular diseases, such as atherothrombosis and restenosis.
NDRG-1（N-myc 下游调节基因 1）是 NDRG 家族的一员，在细胞分化、增殖和应激反应中发挥重要作用。尽管 NDRG1 的表达受到流体剪切力的调节，但它在血管生物学中的作用仍知之甚少。本研究的目的是确定 NDRG1 在血管炎症和重塑中的功能意义。
通过定量聚合酶链反应、western blot 和免疫组织化学，我们证明 NDRG1 的表达在细胞因子刺激的内皮细胞和人及鼠动脉粥样硬化病变中明显增加。为了确定 NDRG1 在血管内皮细胞激活中的作用，我们使用 NDRG1 短发夹 RNA 进行了功能丧失研究。我们的结果表明，通过携带 NDRG1 短发夹 RNA 的慢病毒进行 NDRG1 敲低可显著减弱白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）诱导的细胞因子/趋化因子和黏附分子的表达。有趣的是，抑制 NDRG1 还显著减弱了促凝分子，如纤溶酶原激活物抑制剂 1（PAI-1）和组织因子（TF）的表达，同时增加了血栓调节蛋白（TM）和组织型纤溶酶原激活物（t-PA）的表达，从而在内皮细胞中发挥强大的抗血栓作用。在机制上，我们发现 NDRG1 与孤儿 Nur77（核受体）相互作用，并在血管内皮细胞中抑制 Nur77 和核因子 Kappa B（NF-κB）的转录活性。此外，在 NDRG1 敲低细胞中，细胞因子诱导的丝裂原激活蛋白激酶激活、c-Jun 磷酸化和 AP-1（激活蛋白 1）转录活性均明显受到抑制。与野生型同窝仔相比，内皮细胞特异性 NDRG1 敲除小鼠颈动脉结扎和动脉血栓形成诱导的新生内膜和动脉粥样硬化形成明显减弱。
我们的研究结果首次表明，NDRG1 是一种关键的调节因子，参与调节内皮炎症、血栓形成反应和血管重塑，并提示抑制 NDRG1 可能成为动脉粥样硬化和再狭窄等炎症性血管疾病的一种新的治疗策略。
