Department of Internal Medicine II, Division of Cardiology, Paracelsus Medical University of Salzburg, Salzburg, Austria.
Department of Internal Medicine II, Division of Cardiology, Paracelsus Medical University of Salzburg, Salzburg, Austria.
Life Sci. 2020 Dec 1;262:118474. doi: 10.1016/j.lfs.2020.118474. Epub 2020 Sep 19.
Blood coagulation is one of the most important host-defending mechanisms in vivo by maintaining the blood pressure after injury. However, besides maintaining homeostasis, blood coagulation and the contributing factors are directly linked to pathological conditions, such as thromboembolism and inflammation, leading to cardiovascular diseases, among others. As anti-inflammatory drugs may reduce cardiovascular events, we hypothesized in this study that the direct thrombin inhibitor dabigatran may reduce cytokine, growth factor and chemokine expression in vitro.
Initially, human whole blood was incubated in tubes for serum, EDTA plasma, and heparinized plasma. Furthermore, human PBMCs were isolated and incubated under different culture conditions, including the treatment with human serum or thrombin, respectively. The effect of the oral anticoagulant dabigatran on pro-inflammatory cytokines, growth factors and chemokines was investigated by ELISA.
Conditioned serum resulted in a significant alteration of the secretome's protein levels after 24 h. However, solely ANG showed a dose-dependent increment by the addition of serum (79.8 ± 9.2 ng/mL) in comparison to baseline (0.2 ± 0.2 ng/mL), as it was in trend for thrombin treatment. Furthermore, the pre-treatment of PBMCs with different doses of dabigatran significantly lowered supernatant protein levels measured. Moreover, dabigatran was shown to decrease most notably the growth factor and chemokine levels in the PBMC's secretome that were treated with 200 ng/mL thrombin in a dose-dependent manner.
In conclusion, novel oral anticoagulants, such as dabigatran, could help to reduce not only procoagulatory effects in inflammatory conditions but could also reduce proinflammatory stimuli via reduced expression of cytokines and chemokines.
血液凝固是体内维持受伤后血压的最重要的宿主防御机制之一。然而，除了维持体内平衡外，凝血和促成因素与血栓栓塞和炎症等病理状况直接相关，导致心血管疾病等。由于抗炎药物可能减少心血管事件，我们假设在这项研究中，直接凝血酶抑制剂达比加群可能会减少细胞因子、生长因子和趋化因子的表达。
最初，将人全血孵育在含有血清、EDTA 血浆和肝素化血浆的试管中。此外，分离出人外周血单核细胞，并在不同的培养条件下孵育，包括分别用人类血清或凝血酶处理。通过 ELISA 研究口服抗凝剂达比加群对促炎细胞因子、生长因子和趋化因子的影响。
条件血清在 24 小时后导致分泌组蛋白水平发生显著变化。然而，仅 ANG 在添加血清后呈剂量依赖性增加（79.8±9.2ng/mL），与基线（0.2±0.2ng/mL）相比，与凝血酶处理呈趋势。此外，用不同剂量的达比加群预处理 PBMCs 可显著降低上清液蛋白水平。此外，达比加群在剂量依赖性方式下显著降低用 200ng/mL 凝血酶处理的 PBMC 分泌组中的生长因子和趋化因子水平。
总之，新型口服抗凝剂，如达比加群，不仅可以帮助减少炎症状态下的促凝血作用，还可以通过减少细胞因子和趋化因子的表达来减少促炎刺激。
