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Effect of corticosteroids and long-acting ß2- agonists in a human cell culture based "in vitro" model of airway inflammation and tissue remodeling

Goulet, Stéphanie. Effect of corticosteroids and long-acting ß2- agonists in a human cell culture based "in vitro" model of airway inflammation and tissue remodeling. 2006, Doctoral Thesis, University of Basel, Faculty of Science.

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Official URL: http://edoc.unibas.ch/diss/DissB_7631

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Abstract

Asthma and chronic obstructive pulmonary disease (COPD) are characterized by chronic
airway inflammation and remodeling. In non-pathological conditions tissue homeostasis is
maintained by the balance between extracellular matrix (ECM) synthesis and degradation.
However, this equilibrium is altered in asthma and COPD leading to structural changes such
as increased ECM deposition. Fibroblasts and airway smooth muscle cells (ASMC) are the
main source of collagen and glycosaminoglycans (GAGs) and transforming growth factor-b1
(TGF-b1) is the most potent ECM stimulator in vitro. In turn, the ECM influences cell
proliferation, migration, differentiation, and secretion of cytokines and growth factors. Thus,
ECM deposition may perpetuate the inflammatory and remodeling processes, contributing to
progression and severity of diseases. Inhaled corticosteroids and long acting b2-agonists
(LABA) are the current therapy for asthma and COPD but their effect on airway remodeling
is not clear. Therefore, the purpose of this thesis was to investigate the effect of
corticosteroids and LABA on airway inflammation and tissue remodeling using a human cell
based in vitro model.
In the present thesis, primary human lung fibroblasts and ASMC were cultured under serumfree
condition to reflect a non-inflammatory environment or with 5% fetal calf serum (FCS)
and/or TGF-b1 to mimic inflammation. This experimental design allowed us to assess the
effect of corticosteroids and LABA on total ECM, collagen and GAGs deposition, cell
proliferation, cytokines release, collagen and ECM mediators mRNA expression, and
gelatinolytic activity under both non-inflammatory and inflammatory conditions.
Concerning the effect of corticosteroids and LABA on ECM deposition by fibroblasts, we
showed that 5% FCS and TGF-b1 increased total ECM and collagen deposition. Under serumfree
condition corticosteroids reduced ECM deposition and the effect was partly mediated by
the glucocorticoid receptor and collagen de novo synthesis. Interestingly, in the presence of
5% FCS corticosteroids had the opposite effect. LABA reduced total ECM and collagen
deposition under both conditions. Combined drugs further decreased ECM deposition under
serum-free condition whereas they counteracted each other in 5% FCS, independently of
TGF-b1. Furthermore, we showed that 5% FCS and TGF-b1 increased GAGs secretion and
deposition. Under serum-free condition corticosteroids inhibited GAGs secretion and
deposition whereas LABA alone had no effect but partly reversed the effect of corticosteroids.
On the contrary, in 5% FCS corticosteroids and LABA increased GAGs deposition. Upon
TGF-b1 stimulation, similar effects were obtained except in 5% FCS, where both drugs
decreased the TGF-b1-induced GAGs secretion and deposition. Together, these findings
indicate that the effect of corticosteroids but not of LABA, on ECM deposition by fibroblasts
is altered by ongoing inflammation.
The effects of corticosteroids and LABA on GAGs, matrix metalloproteinases (MMPs) and
total ECM/collagen deposition by ASMC from healthy, asthma and COPD patients were also
investigated. First, no difference in GAGs secretion and deposition was observed between
ASMC from healthy, asthma and COPD patients. Under serum-free condition corticosteroids
inhibited GAGs secretion and deposition while LABA had no clear effect nor modulated the
effect of corticosteroids. In 5% FCS corticosteroids decreased GAGs secretion to a greater
extent than LABA and their combination resulted in the same effect as corticosteroids alone.
None of the drugs had any effect on GAGs deposition. Second, we assessed the effect of the
drugs on ASMC-derived MMPs. In ASMC only proMMP-2 gelatinolytic activity was
detected, which was decreased in asthma and COPD patients compared to healthy controls.
However, corticosteroids and/or LABA had no effect on proMMP-2 activity. Lastly, we
demonstrated that corticosteroids and LABA generated similar effects on total ECM and
collagen deposition by ASMC from healthy subjects as observed with fibroblasts.
In conclusion, our novel findings suggest that the action of corticosteroids and LABA on
ECM deposition differs under non-inflammatory and inflammatory conditions in lung
fibroblasts and ASMC. In the presence of acute inflammation with vessel leakage, increased
ECM deposition may be regarded as an attempt of stromal cells to block further serum and
cells infiltration into the tissue and this effect would be supported by corticosteroids. The
addition of LABA would counteract the corticosteroid-induced ECM deposition and therefore
their combination with corticosteroids may depend on the inflammatory status of the patient.
Thus, when airway inflammation is resolved, combination therapy may beneficially reduce
pathological tissue remodeling. Thus, our findings might have implications for the short- and
long-term treatment strategies in regard to airway remodeling in asthma and COPD.
Advisors:Eberle, Alex N.
Committee Members:Resink, Therese and Ballmer-Hofer, Kurt
Faculties and Departments:03 Faculty of Medicine > Departement Biomedizin > Former Units at DBM > Endocrinology (Eberle)
UniBasel Contributors:Eberle, Alex N. and Ballmer-Hofer, Kurt
Item Type:Thesis
Thesis Subtype:Doctoral Thesis
Thesis no:7631
Thesis status:Complete
Bibsysno:Link to catalogue
Number of Pages:135
Language:English
Identification Number:
Last Modified:05 Apr 2018 17:32
Deposited On:13 Feb 2009 15:42

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