Allergy Asthma Immunol Res.  2016 Mar;8(2):161-169. 10.4168/aair.2016.8.2.161.

Administration of Pigment Epithelium-Derived Factor Inhibits Airway Inflammation and Remodeling in Chronic OVA-Induced Mice via VEGF Suppression

• Zha W
• Su M
• Huang M
• Cai J
• Du Q

• KMID: 2165933
• DOI: http://doi.org/10.4168/aair.2016.8.2.161

Abstract

PURPOSE
Pigment epithelium-derived factor (PEDF) is a recently discovered antiangiogenesis protein. PEDF possesses powerful anti-inflammatory, antioxidative, antiangiogenic, and antifibrosis properties. It has been reported that PEDF can regulate vascular endothelial growth factor (VEGF) expression. This study aimed to evaluate whether recombinant PEDF protein could attenuate allergic airway inflammation and airway remodeling via the negative regulation of VEGF using a murine model of chronic ovalbumin (OVA)-induced asthma and BEAS-2B human bronchial epithelial cells.
METHODS
In an in vivo experiment, mice sensitized with OVA were chronically airway challenged with aerosolized 1% OVA solution for 8 weeks. Treated mice were given injections of recombinant PEDF protein (50 or 100 microg/kg body weight) via the tail vein. In an in vitro experiment, we investigated the effects of recombinant PEDF protein on VEGF release levels in BEAS-2B cells stimulated with IL-1beta.
RESULTS
Recombinant PEDF protein significantly inhibited eosinophilic airway inflammation, airway hyperresponsiveness, and airway remodeling, including goblet cell hyperplasia, subepithelial collagen deposition, and airway smooth muscle hypertrophy. In addition, recombinant PEDF protein suppressed the enhanced expression of VEGF protein in lung tissue and bronchoalveolar lavage fluid (BALF) in OVA-challenged chronically allergic mice. In the in vitro experiment, VEGF expression was increased after IL-1beta stimulation. Pretreatment with 50 and 100 ng/mL of recombinant PEDF protein significantly attenuated the increase in VEGF release levels in a concentration-dependent manner in BEAS-2B cells stimulated by IL-1beta.
CONCLUSIONS
These results suggest that recombinant PEDF protein may abolish the development of characteristic features of chronic allergic asthma via VEGF suppression, providing a potential treatment option for chronic airway inflammation diseases such as asthma.

Keyword

Asthma; airway inflammation; airway remodeling; PEDF; VEGF

MeSH Terms

Airway Remodeling
Animals
Asthma
Bronchoalveolar Lavage Fluid
Collagen
Eosinophils
Epithelial Cells
Goblet Cells
Humans
Hyperplasia
Hypertrophy
Inflammation*
Lung
Mice*
Muscle, Smooth
Ovalbumin
Ovum
Tail
Vascular Endothelial Growth Factor A*
Veins
Collagen
Ovalbumin
Vascular Endothelial Growth Factor A

Figure

• 

  Fig. 1 PEDF suppresses (A) airway hyperreactivity (AHR) and (B) inflammatory cell accumulation in bronchoalveolar lavage fluid (BALF) in a murine model of asthma. (A) PEDF decreased AHR compared to ovalbumin (OVA)-sensitized and challenged mice. (B) PEDF decreased the accumulation of total inflammatory cells, as well as eosinophils, compared to the OVA group. Data are presented as means±SEM (n=8 per group). *P<0.05 compared to the control group; **P<0.05 compared to the OVA group.

• 

  Fig. 2 PEDF attenuates airway inflammation in a murine model of asthma. Hematoxylin and eosin staining (original magnification ×100): (A) control group, (B) OVA group, (C) PEDF low group, and (D) PEDF high group.

• 

  Fig. 3 PEDF inhibits IL-4, IL-5, IL-13, and TGF-β1 in BALF and OVA-specific IgE in serum in a murine model of asthma. Data are presented as means±SEM (n=8 per group). *P<0.05 compared to the control group; **P<0.05 compared to the OVA group.

• 

  Fig. 4 PEDF attenuates airway goblet cell hyperplasia and mucus production in a murine model of asthma. Periodic acid-Schiff (PAS) staining (original magnification ×200): (A) control group, (B) OVA group, (C) PEDF low group, and (D) PEDF high group. (E) Quantitative analyses of mucus production in lung sections were performed as described in the Methods. Data are presented as means±SEM (n=8 per group). *P<0.05 compared to the control group; **P<0.05 compared to the OVA group.

• 

  Fig. 5 PEDF inhibits peribronchiolar fibrosis in chronic experimental asthma. Masson's trichrome-positive peribronchiolar collagen layer (original magnification ×200) of lung tissue from (A) control group, (B) OVA group, (C) PEDFlow group, and (D) PEDFhigh group. (E) Quantitative analyses of the mean area of airway fibrosis and (F) total lung hydroxyproline content were determined as described in the Methods. Data are presented as means±SEM (n=8 per group). *P<0.05 compared to the control group; **P<0.05 compared to the OVA group.

• 

  Fig. 6 PEDF inhibits the areas of α-smooth muscle actin (SMA) in a murine model of asthma. Peribronchial α-SMA immunostaining (immunohistochemistry, original magnification ×200) of lung tissue from (A) control group, (B) OVA group, (C) PEDF low group, and (D) PEDF high group. (E) Quantitative analyses of immunostaining area for peribronchial α-SMA. Data are presented as means±SEM (n=8 per group). *P<0.05 compared to the control group; **P<0.05 compared to the OVA group.

• 

  Fig. 7 PEDF ameliorates the expression of VEGF protein in lung tissue in chronic experimental asthma. (A) Total protein from lung tissue was extracted 24 hours after the final OVA challenge and subjected to Western blot analysis of VEGF. GAPDH was utilized as the standard control. (B) The band signal strength of VEGF expressed as a ratio to GAPDH. Data are presented as means±SEM (n=8 per group). *P<0.05 compared to the control group; **P<0.05 compared to the OVA group.

• 

  Fig. 8 PEDF reduces BALF VEGF levels in chronic experimental asthma. Data are presented as means±SEM (n=8 per group). *P<0.05 compared to the control group; **P<0.05 compared to the OVA group.

• 

  Fig. 9 Recombinant PEDF suppresses IL-1β-induced VEGF release in BEAS-2B cells. Data are presented as means±SEM (n=3 per group). *P<0.05 compared to the control group; **P<0.05 compared to the IL-1β group.

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