Nutr Res Pract.  2007 Dec;1(4):285-290.

Resveratrol blunts tumor necrosis factor-alpha-induced monocyte adhesion and transmigration

• Kim DS
• Kwon HM
• Choi JS
• Kang SW
• Ji GE
• Kang YH

Abstract

The leukocyte recruitment and transmigration across the endothelial barrier into the vessel wall are crucial steps in atherosclerosis. Leukocyte trafficking on the endothelium is elicited by induction of endothelial adhesion molecules, and its transmigration is mediated by degradation of basement membrane proteins through enzymatic activity of matrix metalloproteinases (MMP). The current study investigated whether resveratrol, a polyphenol present in grapes and red wine, was capable of inhibiting leukocyte adhesion to tumor necrosis factor (TNF)-alpha-activated endothelium. It was found that resveratrol inhibited the TNF-alpha-activated endothelial expression of vascular cell adhesion molecule-1 in a dose-dependent manner. In addition, resveratrol hampered THP-1 monocyte adhesion to activated endothelial cells. This study further examined whether resveratrol interfered with transendothelial migration of leukocytes. The MMP-2 gelatinolytic activity of endothelial cells was enhanced by TNF-alpha, which was attenuated by an addition of > or =25 micrometer resveratrol. In addition, 25 micrometer resveratrol mitigated the MMP-9 activity of THP-1 cells, followed by a marked inhibition of transendothelial migration. These results demonstrated that resveratrol suppressed monocyte adhesion and migration induced by TNF-alpha through modulating expression of adhesion molecules and gelatinolytic activity of MMP. These findings suggest that dietary resveratrol may be therapeutic agent for inhibiting leukocyte recruitment into the subendothelium during inflammatory atherosclerosis.

Keyword

Resveratrol; TNF-alpha; VCAM-1; THP-1 monocyte; MMP

MeSH Terms

Atherosclerosis
Basement Membrane
Endothelial Cells
Endothelium
Leukocytes
Matrix Metalloproteinases
Monocytes*
Necrosis*
Transendothelial and Transepithelial Migration
Tumor Necrosis Factor-alpha
Vascular Cell Adhesion Molecule-1
Vitis
Wine
Matrix Metalloproteinases
Tumor Necrosis Factor-alpha
Vascular Cell Adhesion Molecule-1

Figure

• 

  Fig. 1 Inhibitory dose response of THP-1 monocytes adhesion to resveratrol in TNF-α-activated HUVEC. HUVEC were pre-treated at 10-50 µM resveratrol overnight and then activated with 10 ng/mL TNF-α for 6 h. Endothelial cells were co-cultured with calcein AM-labeled THP-1 monocytes for 2 h. Microphotographs (3 independent experiments) were obtained using a fluorescence microscopy. Magnification ×200. The bar graphs (bottom panel) represent quantitative results obtained by using a Fluoroscan ELISA plate reader at λ=485 nm excitation and λ=538 nm emission. *p<0.05, relative to TNF-α-untreated control incubation. +p<0.05, relative to TNF-α-alone incubation.

• 

  Fig. 2 Resveratrol inhibition of VCAM-1 induction in TNF-α-stimulated HUVEC. After culturing HUVEC with 10-50 µM resveratrol and 10 ng/mL TNF-α, cell extracts were subjected to 8% SDS-PAGE, followed by Western blot analysis with a VCAM-1 primary antibody. β-Actin protein was used as an internal control. The bar graphs (bottom panel) represent densitometric results (3 independent experiments). *p<0.05, relative to TNF-α-untreated control incubation. +p<0.05, relative to TNF-α-alone incubation. ++p<0.05, relative to incubation with 25 µM resveratrol.

• 

  Fig. 3 Gelatinolytic activity of MMP-2 (A) and MMP-9 (B) in TNF-α-exposed HUVEC and THP-1 cells. HUVEC (for MMP-2) and THP-1 cells (for MMP-9) were with 25 µM resveratrol overnight prior to an addition of 10 ng/mL TNF-α and continuously incubated for 24 h. The collected culture media were subjected to electrophoresis on 10% SDS-PAGE co-polymerized by 0.1% gelatin as the substrate. Gelatinolytic activities were detected as unstained bands against the background of Coomassie blue-stained gelatin and gel photographs (3 separate experiments) were obtained.

• 

  Fig. 4 Resveratrol inhibition of TNF-α-induced THP-1 monocyte transmigration. THP-1 cells were incubated by serum free-RPMI-1640 on the transwell chamber coated with 0.1% gelatin on its lower surface. Subsequently, cells were incubated for 48 h in the presence of 25 µM resveratrol and 10 ng/mL TNF-α. THP-1 cells migrated to the lower compartment of transwell chamber were microscopically photographed. The THP-1 cell migration ability was defined by the number of THP-1 cells migrated. *p<0.05, relative to TNF-α-untreated control incubation.

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