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.

中文翻译：

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血管miR-181b控制2型糖尿病的组织因子依赖性血栓形成和炎症。

糖尿病的特征在于慢性血管炎症，导致血栓形成全长（f1）组织因子（TF）及其异型剪接（as）TF的病理表达。血源性TF促进因子（F）Xa的产生，导致血栓形成前状态和心血管并发症。MicroRNA（miR）在转录后水平上影响基因表达，并有助于血管内稳态。它们在控制糖尿病相关促凝状态中的独特作用仍然知之甚少。在一组2型糖尿病控制不良的患者中（n = 46），miR-181b的血浆水平与TF通路活性和血管炎症标记物相关。体外，用miR-181b或抗miR-181b转染人类微血管内皮细胞（HMEC）-1和人类单核细胞（THP-1），并使其暴露于肿瘤坏死因子（TNF）α或脂多糖（LPS）。评估了TF同工型，血管黏附分子（VCAM）1和核因子（NF）κB核易位的表达。此外，分析了携带第一个miR-181b基因座缺失的小鼠的主动脉，脾脏，血浆和骨髓源性巨噬细胞（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，这由TF蛋白，TF活性，d-二聚体水平以及血管炎症标志物证明。在HMEC-1中，miR-181b消除了TNFα诱导的flTF，asTF和VCAM1的表达。使用抗miR-181b验证了这些结果。从机制上讲，我们证实了miR-181b介导的importin-α3（KPNA4）抑制作用，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）（单核细胞中TF的主要诱导剂）而降低了LPS对TF同工型和FXa的响应。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，miR-181-/-小鼠的BMDM在LPS刺激下显示出增加的TF表达和FXa产生。miR-181b表观遗传控制糖尿病的促凝状态。