Korean J Physiol Pharmacol.  2015 Jul;19(4):335-340. 10.4196/kjpp.2015.19.4.335.

Zerumbone, Sesquiterpene Photochemical from Ginger, Inhibits Angiogenesis

• Park JH
• Park GM
• Kim JK

• KMID: 2285576
• DOI: http://doi.org/10.4196/kjpp.2015.19.4.335

Abstract

Here, we investigated the role of zerumbone, a natural cyclic sesquiterpene of Zingiber zerumbet Smith, on angiogenesis using human umbilical vein endothelial cells (HUVECs). Zerumbone inhibited HUVECs proliferation, migration and tubule formation, as well as angiogenic activity by rat aorta explants. In particular, zerumbone inhibited phosphorylation of vascular endothelial growth factor receptor-2 and fibroblast growth factor receptor-1, which are key regulators of endothelial cell function and angiogenesis. In vivo matrigel plug assay in mice demonstrated significant decrease in vascularization and hemoglobin content in the plugs from zerumbone-treated mice, compared with control mice. Overall, these results suggest that zerumbone inhibits various attributes of angiogenesis, which might contribute to its reported antitumor effects.

Keyword

Angiogenesis; Zerumbone; Vascular endothelial growth factor receptor; Human umbilical vein endothelial cells

MeSH Terms

Animals
Aorta
Endothelial Cells
Fibroblast Growth Factors
Ginger*
Human Umbilical Vein Endothelial Cells
Mice
Phosphorylation
Rats
Receptors, Vascular Endothelial Growth Factor
Vascular Endothelial Growth Factor Receptor-2
Fibroblast Growth Factors
Receptors, Vascular Endothelial Growth Factor
Vascular Endothelial Growth Factor Receptor-2

Figure

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  Fig. 1 Structure of Zerumbone.

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  Fig. 2 Inhibition of endothelial cell proliferation, migration, and capillary-like tubule formation by zerumbone. HUVECs were plated in 96-well plates, allowed to attach overnight, and then cultured for 24 h with indicated concentration of zerumbone. (A) The proliferation and (B) LDH releases were determined as described in the Methods section. (C) For cell migration, a monolayer of inactivated HUVECs was wounded by scratching with a 0.1 ml pipette tip, and fresh medium containing indicated concentration of zerumbone was added. After 18 h, migration of HUVECs was measured as described in the Methods section. (D) For capillary-like tubule formation, HUVECs were seeded onto matrigel-coated 48-well plates and incubated with indicated concentration of zerumbone for 18 h. Endothelial tubules were photographed and quantitated. The results are means±SEM for at least three independent experiments. ***p<0.001 versus 0 µM zerumbone-treated cells.

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  Fig. 3 Effect of zerumbone on microvessel outgrowth arising from rat aortic rings. Aortic rings isolated from SD rats were embedded in matrigel in 48-well plates, and then fed with medium containing zerumbone for 7 days. Photographs are representative of three independent experiments.

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  Fig. 4 Effects of zerumbone on VEGFR2- and FGFR1-mediated signaling pathways. (A) HUVECs were treated with various concentrations of zerumbone and stimulated with human VEGF (10 ng/ml) for 15 min. Phosphorylation of VEGFR2 by VEGF was determined using Western blot. (B) HUVECs were treated with zerumbone and stimulated with human bFGF (50 ng/ml) for 15 min. Phosphorylation of of FGFR by bFGF were determined using Western blot. The bands were quantified using image analysis software and their relative intensity was expressed as fold against the image of the VEGF- or bFGF- stimulated cells. ***p<0.001 versus VEGF or bFGF+0 µM zerumbone-treated cells.

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  Fig. 5 Effects of zerumbone in vivo angiogenesis by matrigel plug assay. (A) Matrigel containing mouse VEGF (100 ng/ml), mouse bFGF (100 ng/ml), and zerumbone was subcutaneously injected into C57BL/6J mice. After 7 days, matrigels ware removed and photographed. (B) Paraffin sections of matrigel plugs were stained with hematoxylin and eosin. Original magnification, ×100; Scale bar 100 µm. (C) The quantification of neovascularization on the matrigel was performed by measuring the hemoglobin content. The results are reported as the mean±SEM of three independent experiments. Statistical significance is based on the difference when compared with non-treated animals, **p<0.01, ***p<0.001.

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