Department of Clinical Laboratory Science, Kanazawa University Graduate School of Medical Science, Kanazawa, Japan.
Thromb Res. 2012 Sep;130(3):e188-93. doi: 10.1016/j.thromres.2012.07.002. Epub 2012 Jul 20.
Heme oxygenase-1 (HO-1) is the rate limiting enzyme that catalyzes the conversion of heme into biliverdin, free iron, and carbon monoxide (CO). The first human case of HO-1 deficiency showed abnormalities in blood coagulation and the fibrinolytic system. Thus, HO-1 or HO-1 products, such as CO, might regulate coagulation and the fibrinolytic system. This study examined whether tricarbonyldichlororuthenium (II) dimer (CORM-2), which liberates CO, modulates the expression of tissue factor (TF) and plasminogen activator inhibitor type 1 (PAI-1) in human umbilical vein endothelial cells (HUVECs), and TF expression in peripheral blood mononuclear cells (PBMCs). Additionally, we examined the mechanism by which CO exerts its effects.
HUVECs were pretreated with 50 μM CORM-2 for 3 hours, and stimulated with tumor necrosis factor-α (TNF-α, 10 ng/ml) for an additional 0-5 hours. PBMCs were pretreated with 50-100 μM CORM-2 for 1 hour followed by stimulating with lipopolysaccharid (LPS, 10 ng/ml) for additional 0-9 hours. The mRNA and protein levels were determined by RT-PCR and western blotting, respectively.
Pretreatment with CORM-2 significantly inhibited TNF-α-induced TF and PAI-1 up-regulation in HUVECs, and LPS-induced TF expression in PBMCs. CORM-2 inhibited TNF-α-induced activation of p38 MAPK, ERK1/2, JNK, and NF-κB signaling pathways in HUVECs.
CORM-2 suppresses TNF-α-induced TF and PAI-1 up-regulation, and MAPKs and NF-κB signaling pathways activation by TNF-α in HUVECs. CORM-2 suppresses LPS-induced TF up-regulation in PBMCs. Therefore, we envision that the antithrombotic activity of CORM-2 might be used as a pharmaceutical agent for the treatment of various inflammatory conditions.
血红素加氧酶-1（HO-1）是一种限速酶，可催化血红素转化为胆红素、游离铁和一氧化碳（CO）。首例 HO-1 缺乏症患者表现出凝血和纤溶系统异常。因此，HO-1 或其产物，如 CO，可能调节凝血和纤溶系统。本研究检测了三羰基二氯钌（II）二聚体（CORM-2）是否能调节人脐静脉内皮细胞（HUVEC）中组织因子（TF）和纤溶酶原激活物抑制剂 1（PAI-1）的表达，以及外周血单核细胞（PBMC）中 TF 的表达。此外，我们还研究了 CO 发挥作用的机制。
HUVEC 用 50μM CORM-2 预处理 3 小时，再用肿瘤坏死因子-α（TNF-α，10ng/ml）刺激 0-5 小时。PBMC 用 50-100μM CORM-2 预处理 1 小时，然后用脂多糖（LPS，10ng/ml）刺激 0-9 小时。通过 RT-PCR 和 Western blot 分别检测 mRNA 和蛋白水平。
CORM-2 预处理可显著抑制 TNF-α诱导的 HUVEC 中 TF 和 PAI-1 的上调，以及 LPS 诱导的 PBMC 中 TF 的表达。CORM-2 抑制 TNF-α诱导的 HUVEC 中 p38MAPK、ERK1/2、JNK 和 NF-κB 信号通路的激活。
CORM-2 抑制 TNF-α诱导的 TF 和 PAI-1 上调，以及 TNF-α诱导的 MAPKs 和 NF-κB 信号通路的激活。CORM-2 抑制 LPS 诱导的 PBMC 中 TF 的上调。因此，我们设想 CORM-2 的抗血栓活性可用作治疗各种炎症状态的药物。
