Nutr Res Pract.  2011 Oct;5(5):381-388.

Comparative effect of genistein and daidzein on the expression of MCP-1, eNOS, and cell adhesion molecules in TNF-alpha-stimulated HUVECs

• Cho HY
• Park CM
• Kim MJ
• Chinzorig
• Cho CW
• Song YS

Abstract

We compared the effects of genistein and daidzein on the expression of chemokines, cell adhesion molecules (CAMs), and endothelial nitric oxide synthase (eNOS) in tumor necrosis factor (TNF)-alpha-stimulated human umbilical vascular endothelial cells (HUVECs). TNF-alpha exposure significantly increased expression of monocyte chemoattractant protein (MCP)-1, vascular adhesion molecule (VCAM)-1, and intercellular adhesion molecule-1. Genistein significantly decreased MCP-1 and VCAM-1 production in a dose-dependent manner, whereas CAM expression was not significantly lowered by genistein treatment. However, daidzein slightly decreased MCP-1 production. The effects of genistein and daidzein on MCP-1 secretion coincided with mRNA expression. Pre-treatment with either genistein or daidzein elevated eNOS expression and nitric oxide production disturbed by TNF-alpha exposure. A low concentration of isoflavones significantly inhibited nuclear factor (NF)kappaB activation, whereas a high dose slightly ameliorated these inhibitive effects. These results suggest that genistein had a stronger effect on MCP-1 and eNOS expression than that of daidzein. Additionally, NFkappaB transactivation might be partially related to the down-regulation of these mRNAs in TNF-alpha-stimulated HUVECs.

Keyword

Genistein; MCP-1; cell adhesion molecules; eNOS; NFkappaB

MeSH Terms

Cell Adhesion
Cell Adhesion Molecules
Chemokines
Down-Regulation
Endothelial Cells
Genistein
Humans
Intercellular Adhesion Molecule-1
Isoflavones
Monocytes
Nitric Oxide
Nitric Oxide Synthase Type III
RNA, Messenger
Transcriptional Activation
Tumor Necrosis Factor-alpha
Vascular Cell Adhesion Molecule-1
Cell Adhesion Molecules
Chemokines
Genistein
Intercellular Adhesion Molecule-1
Isoflavones
Nitric Oxide
Nitric Oxide Synthase Type III
RNA, Messenger
Tumor Necrosis Factor-alpha
Vascular Cell Adhesion Molecule-1

Figure

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  Fig. 1 Effects of genistein and daidzein on cell viability in tumor necrosis factor (TNF)-α-stimulated human umbilical vascular endothelial cells (HUVECs). Cells (1 × 105 cells) in 96-well plates were preincubated with and without the indicated concentrations of genistein or daidzein for 2 hr and then incubated with TNF-α (10 ng/mL) for 20 hr. Data represent the means ± SD of triplicate experiments. Values sharing the same superscript are not significantly different at P < 0.05.

• 

  Fig. 2 Inhibitory effects of genistein and daidzein on vascular adhesion molecule (VCAM)-1 production in tumor necrosis factor (TNF)-α-stimulated human umbilical vascular endothelial cells (HUVECs). Cells (4 × 105 cells) in 24-well plates were preincubated with and without the indicated concentrations of genistein or daidzein for 2 hr and then incubated with TNF-α (10 ng/mL) for 20 hr. Data represent the means ± SD of triplicate experiments. Values sharing the same superscript are not significantly different at P < 0.05.

• 

  Fig. 3 Inhibitory effects of genistein and daidzein on monocyte chemoattractant protein (MCP)-1 production in tumor necrosis factor (TNF)-α-stimulated human umbilical vascular endothelial cells (HUVECs). Cells (4 × 105 cells) in 24-well plates were preincubated with and without the indicated concentrations of genistein or daidzein for 2 hr and then incubated with TNF-α (10 ng/mL) for 20 hr. Data represent the means ± SD of triplicate experiments. Values sharing the same superscript are not significantly different at P < 0.05.

• 

  Fig. 4 Effects of genistein on mRNA gene expression in tumor necrosis factor (TNF)-α-stimulated human umbilical vascular endothelial cells (HUVECs). Cells (1 × 106 cells) in 100 mm dishes were preincubated with and without the indicated concentrations of genistein for 2 hr and then incubated with TNFα (10 ng/mL) for 20 hr. Untreated represents the negative control without TNF-α treatment. (A) Levels of mRNA were determined by RT-PCR analysis. GAPDH was used as an internal control. (B) All signals were normalized to GAPDH mRNA levels and expressed as ratios. Data represent means ± SD of triplicate experiments. Values sharing the same superscript are not significantly different at P < 0.05.

• 

  Fig. 5 Effects of daidzein on mRNA gene expression in tumor necrosis factor (TNF)-α-stimulated human umbilical vascular endothelial cells (HUVECs). Cells (1 × 106 cells) in 100 mm dishes were preincubated with and without the indicated concentrations of daidzein for 2 hr and then incubated with TNFα (10 ng/mL) for 20 hr. Untreated represents the negative control without TNF-α treatment. (A) Levels of mRNA expression were determined by RT-PCR analysis. GAPDH was used as an internal control. (B) All signals were normalized to GAPDH mRNA levels and expressed as ratios. Data represent the means ± SD of triplicate experiments. Values sharing the same superscript are not significantly different at P < 0.05.

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  Fig. 6 Effects of genistein and daidzein on nitric oxide (NO) production in tumor necrosis factor (TNF)-α-stimulated human umbilical vascular endothelial cells (HUVECs). Cells (4 × 105 cells) in 24-well plates were preincubated with and without the indicated concentrations of genistein or daidzein for 2 hr and then incubated with TNF-α (10 ng/mL) for 20 hr. Data represent the means ± SD of triplicate experiments. Values sharing the same superscript are not significantly different at P < 0.05.

• 

  Fig. 7 Effects of genistein and daidzein on nuclear factor (NF)κB activity in tumor necrosis factor (TNF)-α-stimulated human umbilical vascular endothelial cells (HUVECs). Cells (1 × 106 cells) in 100 mm dishes were preincubated with and without the indicated concentrations of genistein or daidzein for 2 hr and then incubated with TNF-α (10 ng/mL) for 20 hr. (A) DNA binding activity of NFκB was assessed by electrophoretic mobility shift assay (EMSA)(B) Values are expressed as relative intensity of radioactivity. Data represent the means ± SD of triplicate experiments. Values sharing the same superscript are not significantly different at P < 0.05.

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