Department of Cardiology, Nanjing First Hospital, Nanjing Medical University, Nanjing, China.
Department of Cardiology, Nanjing First Hospital, Nanjing Medical University, Nanjing, China.
Exp Cell Res. 2019 Jul 1;380(1):36-46. doi: 10.1016/j.yexcr.2019.02.022. Epub 2019 Feb 22.
Pulmonary arterial hypertension (PAH) is a diffuse pulmonary microvascular remodeling disease accompanied by malignant proliferation of pulmonary artery smooth muscle cells (PASMCs), which causes persistent pulmonary artery pressure elevation, right ventricular hypertrophy (RVH) and death. However, current therapies targeting pulmonary vascular remodeling and RVH remain poorly effective in reversing PAH. Overactivation of the protein tyrosine kinase Src plays an important role in tumor cell growth, proliferation and invasion; we thus hypothesized that inhibitors targeting Src activation could reverse experimental PAH. We demonstrated that Src was markedly activated in hypoxia-stimulated PASMCs from donors and PASMCs isolated from PAH patients. We investigated the effects of the Src-selective inhibitor 1-(1,1-dimethylethyl)-1-(4-methylphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (PP1) and berberine (BBR) on PAH-PASMC proliferation and migration by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), 5-ethynyl-2'-deoxyuridine (EdU) and wound-healing assays. Our in vitro results showed that inhibition of Src (Tyr416) phosphorylation repressed PAH-PASMC proliferation and migration by inhibiting hypoxia-inducible factor-1α (HIF-1α) expression through Akt/mTOR signal pathway. In vivo, PP1 and BBR significantly alleviated distal pulmonary vascular remodeling and decreased right ventricular systolic pressure (RVSP) and RVH in Sugen (SU) 5416/hypoxia (SU-PAH) mice. These findings demonstrate that pharmacological (PP1 or BBR) inhibition of Src activation could be a novel means of treating severe pulmonary vascular remodeling and RVH in PAH patients.
肺动脉高压（PAH）是一种弥漫性肺微血管重塑疾病，伴有肺动脉平滑肌细胞（PASMC）的恶性增殖，导致持续性肺动脉压升高、右心室肥厚（RVH）和死亡。然而，目前针对肺血管重塑和 RVH 的治疗方法在逆转 PAH 方面效果不佳。蛋白酪氨酸激酶Src 的过度激活在肿瘤细胞的生长、增殖和侵袭中起着重要作用；因此，我们假设靶向 Src 激活的抑制剂可以逆转实验性 PAH。我们证明，在缺氧刺激的供体 PASMC 和 PAH 患者分离的 PASMC 中，Src 明显被激活。我们通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）、5-乙炔基-2'-脱氧尿苷（EdU）和划痕愈合试验，研究了Src 选择性抑制剂 1-(1,1-二甲基乙基)-1-(4-甲基苯基)-1H-吡唑并[3,4-d]嘧啶-4-胺（PP1）和小檗碱（BBR）对 PAH-PASMC 增殖和迁移的影响。我们的体外结果表明，Src（Tyr416）磷酸化的抑制通过 Akt/mTOR 信号通路抑制缺氧诱导因子-1α（HIF-1α）的表达，从而抑制 PAH-PASMC 的增殖和迁移。在体内，PP1 和 BBR 显著减轻 SU 5416/缺氧（SU-PAH）小鼠的远端肺血管重塑，降低右心室收缩压（RVSP）和 RVH。这些发现表明，Src 激活的药理学（PP1 或 BBR）抑制可能是治疗 PAH 患者严重肺血管重塑和 RVH 的一种新方法。
