Nutr Res Pract.  2009 Mar;3(1):3-8.

Blockade of vascular angiogenesis by Aspergillus usamii var. shirousamii-transformed Angelicae Gigantis Radix and Zizyphus jujuba

• Kang SW
• Choi JS
• Bae JY
• Li J
• Kim DS
• Kim JL
• Shin SY
• You HJ
• Park HS
• Ji GE
• Kang YH

Abstract

The matrix metalloproteinases (MMP) play an important role in tumor invasion, angiogenesis and inflammatory tissue destruction. Increased expression of MMP was observed in benign tissue hyperplasia and in atherosclerotic lesions. Invasive cancer cells utilize MMP to degrade the extracellular matrix and vascular basement membrane during metastasis, where MMP-2 has been implicated in the development and dissemination of malignancies. The present study attempted to examine the antiangiogenic activity of the medicinal herbs of Aspergillus usamii var. shirousamii-transformed Angelicae Gigantis Radix and Zizyphus jujube (tAgR and tZj) with respect to MMP-2 production and endothelial motility in phorbol 12-myristate 13-acetate (PMA)- or VEGF-exposed human umbilical vein endothelial cells (HUVEC). Nontoxic tAgR and tZj substantially suppressed PMA-induced MMP-2 secretion. In addition, 25 microg/mL tAgR and tZj prevented vascular endothelial growth factor-stimulated endothelial cell transmigration and tube formation. The results reveal that tAgR and tZj dampened endothelial MMP-2 production leading to endothelial transmigration and tube formation. tAgR and tZj-mediated inhibition of endothelial MMP may boost a therapeutic efficacy during vascular angiogenesis.

Keyword

Angiogenesis; Aspergillus. usamii var. shirousamii; transformation; Angelicae Gigantis Radix and Zizyphus jujuba

MeSH Terms

Angelica
Aspergillus
Basement Membrane
Endothelial Cells
Extracellular Matrix
Human Umbilical Vein Endothelial Cells
Hyperplasia
Matrix Metalloproteinases
Neoplasm Metastasis
Phorbols
Plants, Medicinal
Transendothelial and Transepithelial Migration
Ziziphus
Matrix Metalloproteinases
Phorbols

Figure

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  Fig. 1 Cytotoxicity of tAgR (A) and tZj (B) in PMA-exposed HUVEC. After HUVEC were cultured for 24 h in the presence of 50 ng/mL PMA and 1-25 µg/mL tAgR and tZj, MTT assay was performed. The bar graph data represent means ± SEM from 5 independent experiments with multiple estimations. Values are expressed as percent cell survival relative to untreated control cells (cell viability=100%). *P<0.05, compared to untreated control.

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  Fig. 2 Gelatinolytic activity of MMP-2 of PMA-exposed HUVEC treated with 1-25 µg/mL tAgR (A) and tZj (B). After HUVEC culture protocols with 50 ng/mL PMA and 1-25 µg/mL tAgR and tZj for 24 h, equal volumes of culture media were subjected to 10% SDS-PAGE for the measurement of the enzyme activity of MMP-2. Gelatinolytic activity was detected as unstained bands against the background of Coomassie blue-stained gelatin. The bar graphs (means ± SEM, n=4) represent quantitative densitometric results of upper bands. *P<0.05, compared to untreated control. †P<0.05, compared to untreated control and 50 ng/mL PMA.

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  Fig. 3 Inhibition of cell transmigration by tAgR and tZj in VEGF-treated endothelial cells. HUVEC were cultured on gelatin-coated filters of 8 µg/mL pore transwells. Cells transmigrated for 24 h onto the lower surface of the filter were stained with toluidine blue and counted. The bar graphs (means ± SEM, n=4) represent the numbers of cells transmigrated. *P<0.05, compared to untreated control.

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  Fig. 4 Representative images showing tube formation in VEGF-treated endothelial cells plated onto matrigel. After 12 h incubation with 25 µg/mL tAgR and tZj, cells were fixed and images at ×100 magnification were captured. The number of tubes from the images was counted. Multiple five random fields of view were analyzed for the quantitative results. Tube formation results are plotted as means ± SEM (n=4). *P<0.05, compared to VEGF-untreated control.

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