Hematological Research Group, Department of Clinical Medicine, the Faculty of Health Sciences, University of Tromsø, N-9037, Tromsø, Norway.
Thromb Res. 2011 Dec;128(6):e154-9. doi: 10.1016/j.thromres.2011.07.027. Epub 2011 Aug 6.
Bone morphogenetic protein (BMP)-7 regulates atherosclerotic plaque calcification, and it contributes to increased thrombogenicity of lipid-rich lesions by enhancement of TF expression in monocytes/macrophages by unknown mechanism. Since Erk1/2, JNK and p38 mitogen activated protein kinases (MAPKs) regulate TF expression, we studied involvement of MAPK pathways in BMP-7-induced activation of TF in human mononuclear cells (MNCs). Whole blood from healthy volunteers was treated with BMP-7, MNCs were isolated, and TF expression was assessed by western blot (WB) and In-Cell Western assay. Phosphorylation and nuclear translocation of Smad1/5/8 in response to BMP-7 stimulation of MNCs was evaluated by WB and confocal microscopy. Activation of MAPKs was judged by measuring the levels of phoshorylated Erk1/2, JNK, and p38 in the lysates of MNCs. The impact of Erk1/2 and p38 activation was studied by use of PD98059 and SB203580 inhibitors, respectively. Stimulation of whole blood with BMP-7 increased the levels of TF in the lysates of MNCs by 7-fold as compared to 12-fold after LPS stimulation. It was followed by elevation in TF fuctional activity. It was accompanied by elevated levels of phosphorylated Smad 1/5/8 and nuclear translocation of Smad1/5/8 proteins. Treatment of whole blood with BMP-7 led to a phosphorylation of Erk1/2, JNK and p38 MAPKs. BMP-7-induced TF expression was partially inhibited by Erk1/2 inhibitor, whereas TF expression was completely abolished by p38 inhibitor. BMP-7-dependent activation of TF in human MNCs by BMP-7 is accompanied by activation of canonic Smad1/5/8 signaling pathway and depends on activation of Erk1/2 and p38.
骨形态发生蛋白-7（BMP-7）调节动脉粥样硬化斑块钙化，通过未知机制增强单核细胞/巨噬细胞中 TF 的表达，增加富含脂质病变的血栓形成能力。由于 Erk1/2、JNK 和 p38 丝裂原活化蛋白激酶（MAPK）调节 TF 的表达，我们研究了 MAPK 途径在 BMP-7 诱导的人单核细胞（MNC）中 TF 激活中的作用。用 BMP-7 处理健康志愿者的全血，分离 MNC，通过 Western blot（WB）和细胞内 Western 检测法评估 TF 表达。通过 WB 和共聚焦显微镜评估 BMP-7 刺激 MNC 后 Smad1/5/8 的磷酸化和核易位。通过测量 MNC 裂解物中磷酸化 Erk1/2、JNK 和 p38 的水平来判断 MAPK 的激活。分别使用 PD98059 和 SB203580 抑制剂研究 Erk1/2 和 p38 激活的影响。与 LPS 刺激后增加 12 倍相比，用 BMP-7 刺激全血使 MNC 裂解物中的 TF 水平增加了 7 倍。随后，TF 功能活性升高。伴随着磷酸化 Smad 1/5/8 水平的升高和 Smad1/5/8 蛋白的核易位。用 BMP-7 处理全血会导致 Erk1/2、JNK 和 p38 MAPK 的磷酸化。Erk1/2 抑制剂部分抑制 BMP-7 诱导的 TF 表达，而 p38 抑制剂完全抑制 TF 表达。BMP-7 依赖性 TF 在人 MNCs 中的激活伴随着经典 Smad1/5/8 信号通路的激活，并依赖于 Erk1/2 和 p38 的激活。
