Paris Descartes University, Medical School, Assistance Publique Hôpitaux de Paris, Service de Physiologie, EA 2511, Hôpital Cochin, 27 Rue du Faubourg Saint-Jacques, 75014 Paris, France; Clinical and Translational Research Center, Tongji University School of Medicine and Shanghai East Hospital, 150 Jimo Road, Shanghai 200120, China.
Pulm Pharmacol Ther. 2013 Dec;26(6):635-43. doi: 10.1016/j.pupt.2013.07.008. Epub 2013 Aug 6.
Pulmonary hypertension (PH) associated with pulmonary fibrosis (PF) considerably worsens prognosis of interstitial lung diseases (ILD). RhoA/Rho-kinases (ROCK) pathway is implicated in high pulmonary vascular tone and pulmonary fibrosis but the effect of ROCK inhibitors on PH associated with PF is not known. We therefore aimed to determine whether long-term treatment with fasudil, a selective ROCK inhibitor, could attenuate PF and PH induced by bleomycin in mice. Male C57BL/6 mice received a single dose of intratracheal bleomycin (3.3 U/kg) to induce PF. Treatment with fasudil (30 mg kg(-1) day(-1)) was given intraperitoneally for 7, 14 or 21 days until mice underwent hemodynamic measurements. Right ventricular systolic pressure (RVSP) and RV/(LV + S) ratio were assessed. Lung inflammatory cells profiles, including macrophages, neutrophils, lymphocytes B and lymphocytes T were assessed by immunohistochemistry. Lung fibrosis was evaluated by histological and biochemical methods. Pulmonary arteriole muscularization and medial wall thickness (MWT) were evaluated by immunohistochemical staining for α-SMA. Bleomycin induced severe PF and PH in mice, associated with an increased RhoA/ROCK activity in the lung. Fasudil reduced lung inflammation and lung collagen content, and attenuated the increased RVSP, RV hypertrophy, and pulmonary vascular remodeling in bleomycin-intoxicated mice. Fasudil inhibited the increased activity of RhoA/ROCK pathway, and partly altered bleomycin-associated activation of TGF-β1/Smad pathway, via inhibition of Smad2/3 phosphorylation. The efficacy of long-term treatment with fasudil suggests that the blockade of RhoA/ROCK pathway may be a promising therapy for patients with ILD-associated PH.
肺纤维化（PF）相关的肺动脉高压（PH）显著恶化了间质性肺疾病（ILD）的预后。RhoA/Rho 激酶（ROCK）通路与肺血管张力升高和肺纤维化有关，但 ROCK 抑制剂对 PF 相关 PH 的影响尚不清楚。因此，我们旨在确定长期使用 fasudil（一种选择性 ROCK 抑制剂）是否可以减轻博来霉素诱导的小鼠 PF 和 PH。雄性 C57BL/6 小鼠接受单次气管内博来霉素（3.3 U/kg）注射以诱导 PF。fasudil（30 mg/kg/天）通过腹腔内给药，持续 7、14 或 21 天，直到小鼠进行血流动力学测量。评估右心室收缩压（RVSP）和 RV/（LV+S）比值。通过免疫组织化学评估肺炎症细胞谱，包括巨噬细胞、中性粒细胞、B 淋巴细胞和 T 淋巴细胞。通过组织学和生化方法评估肺纤维化。通过α-SMA 免疫组织化学染色评估肺小动脉肌化和中膜厚度（MWT）。博来霉素诱导小鼠发生严重的 PF 和 PH，同时肺中的 RhoA/ROCK 活性增加。fasudil 减少了肺炎症和肺胶原蛋白含量，并减轻了博来霉素中毒小鼠中 RVSP、RV 肥大和肺血管重塑的增加。fasudil 通过抑制 Smad2/3 磷酸化抑制 RhoA/ROCK 通路的增加活性，并且部分改变了博来霉素相关的 TGF-β1/Smad 通路的激活。fasudil 的长期治疗效果表明，阻断 RhoA/ROCK 通路可能是治疗ILD 相关 PH 的一种有前途的方法。
