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Azithromycin Attenuates Fibroblast Growth Factors Induced Vascular Endothelial Growth Factor Via p38MAPK Signaling in Human Airway Smooth Muscle Cells

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Abstract

The airways in asthma and COPD are characterized by an increase in airway smooth muscle (ASM) mass and bronchial vascular changes associated with increased expression of pro-angiogenic growth factors, such as fibroblast growth factors (FGF-1 and FGF-2) and vascular endothelial growth factor (VEGF). We investigated the contribution of FGF-1/-2 in VEGF production in ASM cells and assessed the influence of azithromycin and dexamethasone and their underlying signaling mechanisms. Growth-synchronized human ASM cells were pre-treated with MAPK inhibitors, U0126 for ERK1/2MAPK and SB239063 for p38MAPK as well as with dexamethasone or azithromycin, 30 min before incubation with FGF-1 or FGF-2. Expression of VEGF (VEGF-A, VEGF121, and VEGF165) was assessed by quantitative PCR, VEGF release by ELISA and MAPK phosphorylation by Western blotting. Both FGF-1 and FGF-2 significantly induced mRNA levels of VEGF-A, VEGF121, and VEGF165. The VEGF protein release was increased 1.8-fold (FGF-1) and 5.5-fold (FGF-2) as compared to controls. Rapid transient increase in ERK1/2MAPK and p38MAPK phosphorylation and subsequent release of VEGF from FGF-1 or FGF-2-treated ASM cells were inhibited by respective blockers. Furthermore, azithromycin and dexamethasone significantly reduced both the VEGF release and the activation of p38MAPK pathway in response to FGF-1 or FGF-2 treatment. Our Results demonstrate that FGF-1 and FGF-2 up-regulate VEGF production via ERK1/2MAPK and p38MAPK pathways. Both azithromycin and dexamethasone elicited their anti-angiogenic effects via p38MAPK pathway in vitro, thereby suggesting a possible therapeutic approach to tackle VEGF-mediated vascular remodeling.

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Acknowledgments

GMV is holder of the Glaxo Smith Kline (Belgium) chair in respiratory pharmacology at the Katholieke Universiteit Leuven (KULeuven), and is supported by the Research Foundation Flanders (FWO): G.0643.08 and G.0723.10 and by the onderzoeksfonds KULeuven: OT10/050. BMV is a senior research fellow of the FWO.

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Authors and Affiliations

  1. Laboratory of Pneumology, Katholieke Universiteit Leuven, Leuven, Belgium

    Anna Willems-Widyastuti, Bart M. Vanaudenaerde, Robin Vos, Ellen Dilisen, Stijn E. Verleden, Stéphanie I. De Vleeschauwer, Annemie Vaneylen, Hari S. Sharma & Geert M. Verleden

  2. Department of Pulmonology, Leiden University Medical Center, Leiden, The Netherlands

    Willem I. de Boer

  3. Department of Pathology, VU University Medical Center, Amsterdam, The Netherlands

    Wolter J. Mooi & Hari S. Sharma

  4. Lung Transplantation Unit, University Hospital Gasthuisberg, 49 Herestraat, 3000, Leuven, Belgium

    Geert M. Verleden

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  1. Anna Willems-Widyastuti
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Correspondence to Geert M. Verleden.

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Willems-Widyastuti, A., Vanaudenaerde, B.M., Vos, R. et al. Azithromycin Attenuates Fibroblast Growth Factors Induced Vascular Endothelial Growth Factor Via p38MAPK Signaling in Human Airway Smooth Muscle Cells. Cell Biochem Biophys 67, 331–339 (2013). https://doi.org/10.1007/s12013-011-9331-0

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