Long-term treatment with fasudil improves bleomycin-induced pulmonary fibrosis and pulmonary hypertension via inhibition of Smad2/3 phosphorylation

• Published in: Pulmonary pharmacology & therapeutics 2013-12, Vol.26 (6), p.635-643
• Main Authors: Bei, Yihua, Hua-Huy, Thông, Duong-Quy, Sy, Nguyen, Viet-Ha, Chen, Weihua, Nicco, Carole, Batteux, Frédéric, Dinh-Xuan, Anh Tuan
• Format: Article
• Language: English
• Subjects: 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine - analogs & derivatives; 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine - pharmacology; Animals; Bleomycin; Bleomycin - toxicity; Disease Models, Animal; Humans; Hypertension, Pulmonary - drug therapy; Hypertension, Pulmonary - physiopathology; Immunohistochemistry; Inflammation - drug therapy; Inflammation - physiopathology; Injections, Intraperitoneal; Male; Medical Education; Mice; Mice, Inbred C57BL; Phosphorylation - drug effects; Protein Kinase Inhibitors - pharmacology; Pulmonary fibrosis; Pulmonary Fibrosis - drug therapy; Pulmonary Fibrosis - physiopathology; Pulmonary hypertension; Pulmonary/Respiratory; Rho-kinases; RhoA; Severity of Illness Index; Smad2 Protein - metabolism; Smad3 Protein - metabolism; Time Factors
• ISSN: 1094-5539; 1522-9629
• DOI: 10.1016/j.pupt.2013.07.008

Abstract 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.