Allergy Asthma Immunol Res.  2012 Jan;4(1):37-45. 10.4168/aair.2012.4.1.37.

Role of Angiogenic Factors in Airway Remodeling in an Allergic Rhinitis Murine Model

Affiliations
  • 1Department of Otorhinolaryngology-Head and Neck Surgery, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.
  • 2Department of Otorhinolaryngology, Seoul National University College of Medicine, Seoul, Korea. ygmin312@dreamwiz.com
  • 3Sensory Organ Research Institute, Seoul National University Medical Research Center, Seoul, Korea.
  • 4Department of Immunology, Seoul National University Graduate School of Medicine, Seoul, Korea.
  • 5Department of Otolaryngology-Head and Neck Surgery, Seoul National University Boramae Hospital, Seoul, Korea.
  • 6Department of Otorhinolaryngology, Seoul National University Bundang Hospital, Seoul National University College of Medicine, Seoul, Korea.

Abstract

PURPOSE
There is growing evidence that nasal airway remodeling occurs in allergic rhinitis (AR). Although angiogenesis is an important component of airway remodeling in asthma, its involvement in AR has been little studied. Furthermore, information regarding the role of potent angiogenic factors, such as vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF), in the nasal airway remodeling process is limited. This study was conducted to investigate the role of VEGF and PDGF in nasal airway remodeling, and to assess the preventive effects of anti-angiogenic drugs on this process in a murine AR model.
METHODS
Mice were systemically sensitized and subjected to inhalation of ovalbumin (OVA) twice a week for 3 months. Control mice were challenged with phosphate buffered saline, while the treatment group received SU1498, a VEGF receptor inhibitor, and/or AG1296, a PDGF receptor inhibitor, via intraperitoneal injection 4 hours prior to each OVA inhalation. Staining using hematoxylin and eosin, Masson's trichrome, and periodic acid-Schiff were separately performed to assess eosinophil infiltration, subepithelial fibrosis, and goblet cell hyperplasia, respectively, in the nasal airway. Immunohistochemical staining for matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) was also conducted.
RESULTS
Repetitive intranasal inhalation of OVA resulted in significant increases in eosinophil infiltration, subepithelial fibrosis, goblet cell count, and MMP-9/TIMP-1 expression. Administration of SU1498 or AG1296 prevented these abnormal responses.
CONCLUSIONS
The results of this study suggest that a causal relationship may exist between angiogenic factors and nasal airway remodeling in AR. Inhibition of VEGF or PDGF receptors may, in turn, suppress the remodeling process through the regulation of MMP-9/TIMP-1 expression.

Keyword

Allergic rhinitis; nose; airway remodeling; vascular endothelial growth factor; platelet-derived growth factor

MeSH Terms

Airway Remodeling
Angiogenesis Inducing Agents
Angiogenesis Inhibitors
Animals
Asthma
Cinnamates
Eosine Yellowish-(YS)
Eosinophils
Fibrosis
Goblet Cells
Hematoxylin
Hyperplasia
Inhalation
Injections, Intraperitoneal
Matrix Metalloproteinase 9
Mice
Nose
Ovalbumin
Ovum
Platelet-Derived Growth Factor
Receptors, Platelet-Derived Growth Factor
Receptors, Vascular Endothelial Growth Factor
Rhinitis
Rhinitis, Allergic, Perennial
Tissue Inhibitor of Metalloproteinase-1
Tyrphostins
Vascular Endothelial Growth Factor A
Angiogenesis Inducing Agents
Angiogenesis Inhibitors
Cinnamates
Eosine Yellowish-(YS)
Hematoxylin
Matrix Metalloproteinase 9
Ovalbumin
Platelet-Derived Growth Factor
Receptors, Platelet-Derived Growth Factor
Receptors, Vascular Endothelial Growth Factor
Tissue Inhibitor of Metalloproteinase-1
Tyrphostins
Vascular Endothelial Growth Factor A

Figure

  • Fig. 1 Experimental protocol for mouse sensitization and drug treatment. Mice were sensitized by intraperitoneal injection of inhalation of ovalbumin (OVA) plus aluminum hydroxide on days 0, 7, 14, and 21. Intranasal challenge was performed by daily OVA inhalation on days 28 to 35, followed by drug treatment, and then OVA inhalation was resumed twice a week for the next 3 months. Control mice were sensitized and challenged with PBS twice a week for 3 months. Mice were sacrificed 24 hr after the final OVA challenge. SU1498 or AG1296 treatment was given intraperitoneally 4 hr before each OVA inhalation. Arrows indicate intraperitoneal injection of OVA plus aluminum hydroxide; triangles, intranasal OVA or phosphate-buffered saline challenge; reverse triangles, intraperitoneal SU1498 or AG1296 injection; circles, sacrifice.

  • Fig. 2 Inhalation of ovalbumin (OVA) treatment enhances eosinophil recruitment and the production of IL-4 and TGF-β. (A) Eosinophils were counted in nasal mucosa sections from mice in groups B, C, D1, D2, and D3 that were sacrificed at 0 or 3 months. (B) IL-4 production was measured in the supernatant of OVA-stimulated splenocyte cultures from mice sacrificed at 0 or 3 months. (C) TGF-β production was measured in the supernatant of OVA-stimulated splenocyte cultures from mice sacrificed at 0 or 3 months. Data are presented as the mean±SD. Group A, phosphate-buffered saline treated; group B, OVA treated; group C, dimethyl sulfoxide/OVA treated; group D1, OVA/SU1498 treated; group D2, OVA/AG1296 treated; group D3, OVA/SU1498/AG1296 treated. *P<0.05.

  • Fig. 3 Anti-angiogenic drug treatment significantly reduces subepithelial fibrosis in the nasal septum. Masson's trichrome staining was performed on nasal septum sections of mice at (A) 0 and (B) 3 months. Images were taken at ×200 magnification. (C) A quantitative summary of areas that stained positive for trichrome. Values are reported as percentages of positive-stained area/whole area for each group of mice. Group A, phosphate-buffered saline treated; group B, inhalation of ovalbumin (OVA) treated; group C, dimethyl sulfoxide/OVA treated; group D1, OVA/SU1498 treated; group D2, OVA/AG1296 treated; group D3, OVA/SU1498/AG1296 treated.

  • Fig. 4 Anti-angiogenic drug treatment significantly reduces goblet cell hyperplasia in the nasal septum. Periodic acid-Schiff (PAS) staining was performed on nasal septum sections of mice at (A) 0 and (B) 3 months. Images were taken at ×400 magnification. (C) Goblet cell counts are reported as the mean±SD for each group of mice. Group A, phosphate-buffered saline treated; group B, inhalation of ovalbumin (OVA) treated; group C, dimethyl sulfoxide/OVA treated; group D1, OVA/SU1498 treated; group D2, OVA/AG1296 treated; group D3, OVA/SU1498/AG1296 treated.

  • Fig. 5 Anti-angiogenic drug treatment significantly reduces matrix metalloproteinase-9 (MMP-9) expression in the nasal septum. Nasal septum sections harvested from mice at (A) 0 or (B) 3 months were subjected to immunohistochemical staining for MMP-9, which was detected using confocal microscopy. Images were taken at ×400 magnification. Blue, DAPI; red, MMP-9. (C) Areas that stained positive for MMP-9 are reported for each group of mice as percentages of positive-stained area/whole area. Group A, phosphate-buffered saline treated; group B, inhalation of ovalbumin (OVA) treated; group C, dimethyl sulfoxide/OVA treated; group D1, OVA/SU1498 treated; group D2, OVA/AG1296 treated; group D3, OVA/SU1498/AG1296 treated.

  • Fig. 6 Anti-angiogenic drug treatment significantly reduces tissue inhibitor of metalloproteinase-1 (TIMP-1) expression in the nasal septum. Nasal septum sections harvested from mice at (A) 0 or (B) 3 months were subjected to immunohistochemical staining for TIMP-1, which was detected using confocal microscopy. Images were taken at ×400 magnification. Blue, 4',6-diamidino-2-phenylindole; red, TIMP-1. (C) Areas that stained positive for TIMP-1 are reported for each group of mice as percentages of positive-stained area/whole area. Group A, phosphate-buffered saline treated; group B, inhalation of ovalbumin (OVA) treated; group C, dimethyl sulfoxide/OVA treated; group D1, OVA/SU1498 treated; group D2, OVA/AG1296 treated; group D3, OVA/SU1498/AG1296 treated. *P<0.05.


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