The Effects of Direct Thrombin Inhibition with Dabigatran on Plaque Formation and Endothelial Function in Apolipoprotein E-Deficient Mice
The Effects of Direct Thrombin Inhibition with Dabigatran on Plaque Formation and Endothelial Function in Apolipoprotein E-Deficient Mice
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DOI：
10.1124/jpet.112.194837
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发表时间：
2012-11-01
影响因子：
3.5
通讯作者：
Boehm, Michael
中科院分区：
文献类型：
--
作者：
Lee, Illkyu-O.;Kratz, Mario T.;Boehm, Michael
最近开发的口服抗凝剂达比加群（达比）酯在血浆酯酶活化为达比加群后直接抑制凝血酶。凝血酶参与动脉粥样硬化的发病机制。我们研究了直接凝血酶抑制对加速动脉粥样硬化的高胆固醇血症小鼠模型{[载脂蛋白E缺陷（ApoE（-/-）]小鼠}动脉粥样硬化和内皮功能的影响。ApoE（-/-）小鼠接受富含胆固醇的饲料和达比加群酯（900 mg/kg体重）或溶剂处理12周。野生型（C57/B6）小鼠作为对照。在主动脉环中使用三硝酸甘油（内皮非依赖性）作为对照，用卡巴胆碱（内皮依赖性）评估内皮功能。采用油红染色法观察动脉粥样硬化病变形成情况，天狼星红染色法测定血管胶原含量。活性氧（ROS）的产生通过半定量免疫组织化学染色。达比加群血浆水平测量（622.3 +/- 169 ng/ml）和凝血试验（稀释凝血酶时间）显示相关抗凝血药浓度。达比加群酯减弱了动脉粥样硬化斑块形成增加[ApoE（-/-）达比加群酯：ApoE（-/-）对照的16.1 +/- 3.8%; p < 0.001]、胶原蛋白含量减少[ApoE（-/-）达比加群酯：ApoE（-/-）对照的49.1 +/- 10%; p = 0.01]和二氢乙锭染色中的ROS生成[ApoE（-/-）达比加群酯：ApoE（-/-）对照的46.3 +/- 5.4%; p = 0.005]，与100 μ mol卡巴胆碱时内皮功能的改善平行[ApoE（-/-）对照42.6 +/- 2.7 vs ApoE（-/-）达比加群酯62.9 +/- 3.3%苯肾上腺素诱导的收缩; p = 0.001]。这些数据表明，在高胆固醇血症动脉粥样硬化中，相关剂量的直接凝血酶抑制剂改善了内皮功能，降低了动脉粥样硬化病变的大小、胶原含量和氧化应激。干扰凝血系统可能提供一个治疗靶点，以改变动脉粥样硬化疾病的进展。
The recently developed oral anticoagulant dabigatran (Dabi) etexilate directly inhibits thrombin after activation by plasma esterases to dabigatran. Thrombin is involved in the pathogenesis of atherosclerosis. We investigated the effects of direct thrombin inhibition on atherosclerosis and endothelial function in a hypercholesterolemic mouse model with accelerated atherosclerosis {[apolipoprotein E-deficient (ApoE(-/-)] mice}. ApoE(-/-) mice were treated with a cholesterol-rich diet for 12 weeks and either dabigatran etexilate (900 mg/kg body weight) or vehicle. Wild-type (C57/B6) mice served as control. Endothelial function was assessed with carbachol (endothelium dependent) by using glyceroltrinitrate (endothelium independent) as control in aortic rings. Atherosclerotic lesion formation was evaluated with Oil Red staining, and vascular collagen content was determined by Sirius Red staining. Reactive oxygen species (ROS) production was determined by semiquantitative immunohistochemical staining. Measurement of dabigatran plasma levels (622.3 +/- 169 ng/ml) and a performed coagulation test (diluted thrombin time) revealed a relevant anticoagulatory concentration. Dabigatran etexilate attenuated increased atherosclerotic plaque formation [ApoE(-/-) Dabi: 16.1 +/- 3.8% of ApoE(-/-) control; p < 0.001], decreased collagen content [ApoE(-/-) Dabi: 49.1 +/- 10% of ApoE(-/-) control; p = 0.01], and ROS production in dihydroethidium staining [ApoE(-/-) Dabi: 46.3 +/- 5.4% of ApoE(-/-) control; p = 0.005] in parallel to an improvement of endothelial function [ApoE(-/-) control 42.6 +/- 2.7 versus ApoE(-/-) Dabi 62.9 +/- 3.3% of phenylephrine-induced contraction; p = 0.001] at 100 mu mol carbachol. These data suggest that direct thrombin inhibition in a relevant dosage improved endothelial function and reduced atherosclerotic lesion size, collagen content, and oxidative stress in hypercholesterolemic atherosclerosis. Interference with the coagulation system might provide a therapeutic target to modify atherosclerotic disease progression.