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Basic Science Investigation
Respiration 2018;95:122–136
DOI: 10.1159/000481201
Highly Selective Endothelin-1 Receptor A
Inhibition Prevents Bleomycin-Induced
Pulmonary Inflammation and Fibrosis in Mice
Nikolaos Manitsopoulos
a
Ioanna Nikitopoulou
a
Nikolaos A. Maniatis
a, c
Christina Magkou
d
Anastasia Kotanidou
a, b
Stylianos E. Orfanos
a, c
a
GP Livanos and M Simou Laboratories, 1st Department of Critical Care Medicine & Pulmonary Services, Evangelismos
Hospital, University of Athens Medical School,
b
1st Department of Critical Care Medicine & Pulmonary Services,
Evangelismos Hospital, University of Athens Medical School,
c
2nd Department of Critical Care, “Attikon” Hospital,
University of Athens Medical School, and
d
Department of Pathology, Evangelismos Hospital, Athens, Greece
collagen deposition. Pretreatment with highly selective ETA
receptor inhibitor had no adverse effect on control mice but
improved lung mechanics and lung injury score in addition
to decreasing BALF pleocytosis, protein content, and colla-
gen deposition in BLM-treated mice. Mortality from BLM
reached 40% and occurred primarily during the inflamma-
tory stage of the model but was abrogated by sitaxentan
pretreatment. Conclusions: We conclude that in our BLM-
induced pulmonary fibrosis model, prophylactic highly se-
lective ETA inhibition improves survival, preserves lung func-
tion, attenuates lung injury, and reduces collagen deposi-
tion. © 2017 S. Karger AG, Basel
Introduction
Idiopathic pulmonary fibrosis (IPF), a chronic and
progressive scarring process of the lung [1], is associated
with a poor prognosis, with the majority of patients dying
of respiratory failure within 2–4 years from diagnosis [2,
3]. The pathogenesis of IPF is not fully understood. The
Keywords
Inflammation · Fibrosis · Bleomycin · Endothelin-1 ·
Sitaxentan · Lung function
Abstract
Background: Pulmonary fibrosis is a chronic disease, which
progressively leads to respiratory failure and ultimately
death. Endothelin-1 (ET-1), a vasoconstrictor secreted by en-
dothelial cells, promotes vasoconstriction by activation of its
receptors A and B. Objectives: We addressed the role of
highly selective ET-1 receptor A (ETA) inhibition in the patho-
genesis of experimental pulmonary fibrosis by bleomycin
(BLM). Methods: BLM sulfate (2 U/mL) or saline was intratra-
cheally administered to C57/Bl6 mice (4 groups; n = 5–11/
group). Pretreatment with the highly selective ETA receptor
inhibitor sitaxentan (15 mg/kg/day) was started 1 day prior
to BLM injection and continued for the duration of the ex-
periment. Lung mechanics were assessed prior to sacrifice at
days 7, 14, and 21 after BLM, followed by procurement of
bronchoalveolar lavage fluid (BALF), blood, and lung tissue
samples. Results: Time-dependent effects of BLM exposure
included decreased static compliance and increased lung
elastance, airspace inflammation and microvascular perme-
ability, histological acute lung injury and fibrosis, and lung
Received: February 16, 2017
Accepted after revision: August 30, 2017
Published online: November 9, 2017
Dr. Stylianos E. Orfanos
2nd Department of Critical Care, Attikon University Hospital
1, Rimini Street
Haidari Athens 12462 (Greece)
E-Mail stylianosorfanosuoa @ gmail.com
© 2017 S. Karger AG, Basel
www.karger.com/res
N. Manitsopoulos and I. Nikitopoulou contributed equally to this
work.