Yonsei Med J.  2008 Oct;49(5):804-810. 10.3349/ymj.2008.49.5.804.

The Expression of Adiponectin Receptors and the Effects of Adiponectin and Leptin on Airway Smooth Muscle Cells

• Shin JH
• Kim JH
• Lee WY
• Shim JY

• KMID: 2158199
• DOI: http://doi.org/10.3349/ymj.2008.49.5.804

Abstract

PURPOSE
Obesity is a major risk factor for asthma and it influences airway smooth muscle function and responsiveness. Adiponectin is inversely associated with obesity and its action is mediated through at least 2 cell membrane receptors (AdipoR1 and AdipoR2). Leptin is positively associated with obesity. We investigated whether human airway smooth muscle (ASM) cells express adiponectin receptors and whether adiponectin and leptin regulate human ASM cell proliferation and vascular endothelial growth factor (VEGF) release. MATERIALS AND METHODS: Human ASM cells were growth-arrested in serum-deprived medium for 48 hours and then stimulated with PDGF, adiponectin and leptin. After 48 hours of stimulation, proliferation was determined using a cell proliferation ELISA kit. Human AdipoR1 and -R2 mRNA expressions were determined by RT-PCR using human-specific AdipoR1 and -R2 primers. Concentrations of VEGF, monocyte chemotactic protein (MCP)-1 and macrophage inflammatory protein (MIP)-1alpha in cell culture supernatant were determined by ELISA. RESULTS: Both AdipoR1 and AdipoR2 mRNA were expressed in the cultured human ASM cells. However, adiponectin did not suppress PDGF-enhanced ASM cell proliferation, nor did leptin promote ASM cell proliferation. Leptin promoted VEGF release by human ASM cells, while adiponectin did not influence VEGF release. Neither leptin nor adiponectin influenced MCP-1 secretion from human ASM cells. Adiponectin and MIP-1alpha were not secreted by human ASM cells. CONCLUSION: Human ASM cells expressed adiponectin receptors. However, adiponectin did not regulate human ASM cell proliferation or VEGF release, while leptin stimulated VEGF release by human ASM cells.

Keyword

Smooth muscle cells; cell proliferation; vascular endothelial growth factor; leptin; adiponectin; receptors

MeSH Terms

Adiponectin/metabolism/*pharmacology/physiology
Cell Proliferation/*drug effects
Cells, Cultured
Chemokine CCL2/metabolism
Chemokine CCL3/metabolism
Humans
Leptin/metabolism/*pharmacology/physiology
Myocytes, Smooth Muscle/cytology/drug effects/*metabolism
Obesity/metabolism
Platelet-Derived Growth Factor/metabolism
Receptors, Adiponectin/*metabolism
Respiratory System/cytology/metabolism
Vascular Endothelial Growth Factor A/metabolism

Figure

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  Fig. 1 Expression of adiponectin receptors AdipoR1 and AdipoR2 in the human airway smooth muscle cell. Expression of β-actin (lane 1), AdipoR1 (lane 2) and AdipoR2 (lane 3) mRNA is observed. Sequencing of these PCR products demonstrated that the products were compatible with human AdipoR1 and AdipoR2 (according to NCBI nucleotide-nucleotide BLAST); these findings confirmed the presence of these genes in the human airway smooth muscle cells.

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  Fig. 2 Effect of leptin on the proliferation of human ASM cells. Human ASM cells were stimulated with leptin (10, 50 and 100 ng/mL) and 20 ng/mL of PDGF, and compared to unstimulated cells. Leptin did not enhance or suppress ASM cell proliferation, while PDGF significantly augmented cell stimulation. ASM, airway smooth muscle; PDGF, platelet-derived growth factor. *p < 0.01 vs. Control.

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  Fig. 3 Effect of adiponectin on the proliferation of human ASM cells. Human ASM cells were stimulated with 20 ng/mL of PDGF in the presence or absence of adiponectin (10, 50, and 100 ng/mL), and proliferation was compared with the control. PDGF-stimulated ASM cells showed enhanced proliferation, however, adiponectin did not enhance or suppress PDGF-enhanced ASM cell proliferation. ASM, airway smooth muscle; PDGF, platelet-derived growth factor. *p < 0.01 vs. Control.

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  Fig. 4 Effect of PDGF, adiponectin and leptin on VEGF release by human ASM cells. Human ASM cells were treated with 20 ng/mL of PDGF, 50 ng/mL of adiponectin and 50 ng/mL of leptin. VEGF release was significantly enhanced by PDGF stimulation compared to no stimulation. Leptin significantly augmented VEGF release compared to no stimulation or stimulation by PDGF. Adiponectin showed no stimulatory or inhibitory effects on VEGF release, when it stimulated cells with and without PDGF or leptin. PDGF, platelet-derived growth factor; VEGF, vascular endothelial growth factor. *p < 0.05 vs. Control, **p < 0.05 vs. Control and PDGF.

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  Fig. 5 Effect of PDGF, adiponectin and leptin on MCP-1 release by human ASM cells. Human ASM cells were treated with PDGF (20 ng/mL), adiponectin (50 ng/mL), and leptin (50 ng/mL). MCP-1 was constitutively released from human ASM cells and was neither augmented by PDGF or leptin stimulation, nor suppressed by adiponectin. PDGF, platelet-derived growth factor; ASM, airway smooth muscle.

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