J Korean Med Sci.  2008 Oct;23(5):857-863. 10.3346/jkms.2008.23.5.857.

The Effect of Interleukin-4 and Amphiregulin on the Proliferation of Human Airway Smooth Muscle Cells and Cytokine Release

• Shim JY
• Park SW
• Kim DS
• Shim JW
• Jung HL
• Park MS

• KMID: 1783073
• DOI: http://doi.org/10.3346/jkms.2008.23.5.857

Abstract

Airway smooth muscle (ASM) hyperplasia and angiogenesis are important features associated with airway remodeling. We investigated the effect of IL-4 and amphiregulin, an epidermal growth factor family member, on the proliferation of human ASM cells and on the release of vascular endothelial growth factor (VEGF) and monocyte chemotactic protein (MCP)-1 from human ASM cells. Human ASM cells were growth-arrested for 48 hr and incubated with platelet-derived growth factor (PDGF)- BB, interleukin (IL)-4, amphiregulin, and VEGF to evaluate cell proliferation. The cells were treated with PDGF, IL-4 and amphiregulin to evaluate the release of VEGF, MCP-1. IL-4 suppressed unstimulated and PDGF-stimulated ASM cell proliferation. Amphiregulin stimulated ASM cell proliferation in a dose-dependent manner. VEGF did not have any influence on ASM cell proliferation. IL-4 stimulated VEGF secretion by the ASM cells in a dose-dependent manner and showed added stimulatory effects when co-incubated with PDGF. Amphiregulin did not promote VEGF secretion. IL-4 and amphiregulin showed no stimulatory effects on MCP-1 secretion. The results of this study showed that IL-4 had bifunctional effects on airway remodeling, one was the suppression of the proliferation of the ASM cells and the other was the promotion of VEGF release by the ASM cells, and amphiregulin can promote human ASM cell proliferation.

Keyword

Interleukin-4; Remodeling; Humans, Bronchi; Myocytes, Smooth Muscle; Cell Proliferation; Vascular Endothelial Growth Factor; Amphiregulin

MeSH Terms

Bronchi/metabolism
Cell Proliferation
Cells, Cultured
Chemokine CCL2/metabolism
Chemokine CCL3/metabolism
Cytokines/metabolism
*Gene Expression Regulation
Glycoproteins/*physiology
Humans
Intercellular Signaling Peptides and Proteins/*physiology
Interleukin-4/metabolism/*physiology
Models, Biological
Myocytes, Smooth Muscle/*metabolism
Vascular Endothelial Growth Factor A/metabolism

Figure

• 

  Fig. 1 Effect of IL-4 on the proliferation of human ASM cells. (A) When cells were treated with IL-4 (10, 50, and 100 ng/mL), cell proliferation was significantly suppressed at all concentrations compared to control cells. *p<0.001 vs. Control and PDGF. (B) When cells were treated with IL-4 (10, 100 ng/mL) in presence of PDGF, IL-4 inhibited PDGF-enhanced cell proliferation. †p<0.001 vs. PDGF.

• 

  Fig. 2 Effect of amphiregulin on the proliferation of human ASM cells. (A) Amphiregulin induced cell proliferation in a dose-dependent manner. Amphiregulin significantly enhanced cell proliferation at 50 and 100 ng/mL. *p<0.01 vs. Control. (B) Amphiregulin was added in combination with PDGF to compare the proliferation of cells treated with PDGF only. Proliferation of cells treated with amphiregulin and PDGF did not show an add-on effect compared to the proliferation of cells treated with PDGF only. *p<0.01 vs. Control.

• 

  Fig. 3 (A) Effect of VEGF on the proliferation of human ASM cells. VEGF did not enhance cell proliferation at 10, 30, and 50 ng/mL. (B) Effect of anti-VEGF antibody and anti-VEGF R2 antibody on the PDGF-enhanced cell proliferation. Both 100 ng/mL of anti-VEGF antibody and/or 100 ng/mL of anti-VEGF R2 antibody did not influence PDGF-augmented cell proliferation. VEGF did not augment PDGF-enhanced cell proliferation when cells were treated with 10 ng/mL of VEGF in combination with PDGF. *p<0.01 vs. Control.

• 

  Fig. 4 Effect of IL-4 and amphiregulin on VEGF release by human ASM cells. (A) IL-4 (10 and 100 ng/mL) enhanced VEGF release in a dose-dependent manner. PDGF (20 ng/mL) also significantly increased VEGF release. *p<0.001 vs. Control, IL4-100 and PDGF. (B) IL-4 (10 and 100 ng/mL) increased PDGF-enhanced VEGF secretion. *p<0.01 vs. Control and PDGF. †p<0.01 vs. Control. (C) Amphiregulin (50, 100, and 150 ng/mL) did not augment VEGF secretion at all concentrations. *p<0.001 vs. Control, AR50, AR100 and AR150.

• 

  Fig. 5 Effect of IL-4 and amphiregulin on MCP-1 release by human ASM cells. PDGF (20 ng/mL) increased MCP-1 release. However, neither IL-4 (50 and 100 ng/mL) nor amphiregulin (50 and 100 ng/mL) augmented MCP-1 release by human ASM cells. *p<0.05 vs. Control.

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