Charité Centrum 11, Department of Cardiology, Charité - Universitätsmedizin Berlin, Campus Benjamin Franklin, Hindenburgdamm 30, 12200, Berlin, Germany.
Research Centre Immunology and Institute of Medical Microbiology and Hygiene, University of Mainz Medical Centre, Mainz, Germany.
Cardiovasc Diabetol. 2020 Feb 17;19(1):20. doi: 10.1186/s12933-020-0993-z.
Diabetes mellitus is characterized by chronic vascular inflammation leading to pathological expression of the thrombogenic full length (fl) tissue factor (TF) and its isoform alternatively-spliced (as) TF. Blood-borne TF promotes factor (F) Xa generation resulting in a pro-thrombotic state and cardiovascular complications. MicroRNA (miR)s impact gene expression on the post-transcriptional level and contribute to vascular homeostasis. Their distinct role in the control of the diabetes-related procoagulant state remains poorly understood.
In a cohort of patients with poorly controlled type 2 diabetes (n = 46) plasma levels of miR-181b were correlated with TF pathway activity and markers for vascular inflammation. In vitro, human microvascular endothelial cells (HMEC)-1 and human monocytes (THP-1) were transfected with miR-181b or anti-miR-181b and exposed to tumor necrosis factor (TNF) α or lipopolysaccharides (LPS). Expression of TF isoforms, vascular adhesion molecule (VCAM) 1 and nuclear factor (NF) κB nuclear translocation was assessed. Moreover, aortas, spleen, plasma, and bone marrow-derived macrophage (BMDM)s of mice carrying a deletion of the first miR-181b locus were analyzed with respect to TF expression and activity.
In patients with type 2 diabetes, plasma miR-181b negatively correlated with the procoagulant state as evidenced by TF protein, TF activity, D-dimer levels as well as markers for vascular inflammation. In HMEC-1, miR-181b abrogated TNFα-induced expression of flTF, asTF, and VCAM1. These results were validated using the anti-miR-181b. Mechanistically, we confirmed a miR-181b-mediated inhibition of importin-α3 (KPNA4) leading to reduced nuclear translocation of the TF transcription factor NFκB. In THP-1, miR-181b reduced both TF isoforms and FXa generation in response to LPS due to targeting phosphatase and tensin homolog (PTEN), a principal inducer for TF in monocytes. Moreover, in miR-181-/- animals, we found that reduced levels of miR-181b were accompanied by increased TF, VCAM1, and KPNA4 expression in aortic tissue as well as increased TF and PTEN expression in spleen. Finally, BMDMs of miR-181-/- mice showed increased TF expression and FXa generation upon stimulation with LPS.
miR-181b epigenetically controls the procoagulant state in diabetes. Reduced miR-181b levels contribute to increased thrombogenicity and may help to identify individuals at particular risk for thrombosis.
糖尿病的特征是慢性血管炎症，导致全长（fl）组织因子（TF）及其选择性剪接（as）TF 的病理性表达。血液来源的 TF 促进因子（F）Xa 的产生，导致血栓形成状态和心血管并发症。微小 RNA（miR）在转录后水平影响基因表达，并有助于血管内稳态。它们在控制与糖尿病相关的促凝状态中的独特作用仍知之甚少。
在一组控制不佳的 2 型糖尿病患者（n=46）中，血浆 miR-181b 水平与 TF 途径活性和血管炎症标志物相关。在体外，将人微血管内皮细胞（HMEC-1）和人单核细胞（THP-1）用 miR-181b 或抗 miR-181b 转染，并暴露于肿瘤坏死因子（TNF）α或脂多糖（LPS）。评估 TF 同工型、血管细胞黏附分子（VCAM）1 和核因子（NF）κB 核易位的表达。此外，分析携带第一个 miR-181b 基因座缺失的小鼠的主动脉、脾脏、血浆和骨髓来源的巨噬细胞（BMDM）的 TF 表达和活性。
在 2 型糖尿病患者中，血浆 miR-181b 与促凝状态呈负相关，这可通过 TF 蛋白、TF 活性、D-二聚体水平以及血管炎症标志物来证明。在 HMEC-1 中，miR-181b 阻断了 TNFα 诱导的 flTF、asTF 和 VCAM1 的表达。这些结果使用抗 miR-181b 得到了验证。通过机制研究，我们证实 miR-181b 通过抑制导入蛋白-α3（KPNA4）导致 TF 转录因子 NFκB 的核易位减少。在 THP-1 中，miR-181b 由于靶向磷酸酶和张力蛋白同源物（PTEN），降低了 LPS 诱导的两种 TF 同工型和 FXa 的产生，PTEN 是单核细胞中 TF 的主要诱导物。此外，在 miR-181-/-动物中，我们发现降低的 miR-181b 水平伴随着主动脉组织中 TF、VCAM1 和 KPNA4 表达的增加，以及脾脏中 TF 和 PTEN 表达的增加。最后，LPS 刺激时，miR-181-/-小鼠的 BMDM 显示 TF 表达和 FXa 生成增加。
miR-181b 表观遗传控制糖尿病中的促凝状态。降低的 miR-181b 水平有助于增加血栓形成性，并可能有助于确定血栓形成风险特别高的个体。
