Long-term treatment with fasudil improves bleomycin-induced pulmonary fibrosis and pulmonary hypertension via inhibition of Smad2/3 phosphorylation.
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.
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.
Yihua
Y
Bei
Thông
T
Hua-Huy
Sy
S
Duong-Quy
Viet-Ha
VH
Nguyen
Weihua
W
Chen
Carole
C
Nicco
Frédéric
F
Batteux
Anh Tuan
AT
Dinh-Xuan
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D001761:Bleomycin / Q000633:toxicity; D004195:Disease Models, Animal; D006801:Humans; D006976:Hypertension, Pulmonary / Q000188:drug therapy* / Q000503:physiopathology; D007150:Immunohistochemistry; D007249:Inflammation / Q000188:drug therapy / Q000503:physiopathology; D007274:Injections, Intraperitoneal; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010766:Phosphorylation / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011658:Pulmonary Fibrosis / Q000188:drug therapy* / Q000503:physiopathology; D012720:Severity of Illness Index; D051899:Smad2 Protein / Q000378:metabolism; D051900:Smad3 Protein / Q000378:metabolism; D013997:Time Factors
D047428:Protein Kinase Inhibitors; D051899:Smad2 Protein; C497783:Smad2 protein, mouse; D051900:Smad3 Protein; C497791:Smad3 protein, mouse; D001761:Bleomycin; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; C049347:fasudil
BALF; BLM; Bleomycin; CTD; ILD; IPF; MWT; PF; PH; PVR; Pulmonary fibrosis; Pulmonary hypertension; ROCK; RVSP; Rho-kinase; Rho-kinases; RhoA; bleomycin; bronchoalveolar lavage fluid; connective tissue disease; idiopathic pulmonary fibrosis; interstitial lung disease; medial wall thickness; pulmonary fibrosis; pulmonary hypertension; pulmonary vascular resistance; right ventricular systolic pressure