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An ethanol root extract of Cynanchum wilfordii containing acetophenones suppresses the expression of VCAM-1 and ICAM-1 in TNF-α-stimulated human aortic smooth muscle cells through the NF-κB pathway.

Koo HJ, Sohn EH, Pyo S, Woo HG, Park DW, Ham YM, Jang SA, Park SY, Kang SC - Int. J. Mol. Med. (2015)

Bottom Line: In the present study, we evaluated the anti-inflammatory effects of an ethanol root extract of C. wilfordii (CWE) on tumor necrosis factor (TNF)-α-stimulated human aortic smooth muscle cells (HASMCs).In order to identify the active components in CWE, we re-extracted CWE using several solvents, and found that the ethyl acetate fraction was the most effective in suppressing the expression of VCAM-1 and ICAM-1.We assessed and determined the amounts of these two active components from CWE, and our results suggested that the root of C. wilfordii and its two bioactive acetophenones may be used for the prevention and treatment of atherosclerosis and vascular inflammatory diseases.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicinal and Industrial Crops, Korea National College of Agriculture and Fisheries, Jeonju 560-500, Republic of Korea.

ABSTRACT
The root of Cynanchum wilfordii (C. wilfordii) contains several biologically active compounds which have been used as traditional medicines in Asia. In the present study, we evaluated the anti-inflammatory effects of an ethanol root extract of C. wilfordii (CWE) on tumor necrosis factor (TNF)-α-stimulated human aortic smooth muscle cells (HASMCs). The inhibitory effects of CWE on vascular cell adhesion molecule (VCAM)-1 expression under an optimum extraction condition were examined. CWE suppressed the expression of VCAM-1 and ICAM-1 and the adhesion of THP-1 monocytes to the TNF-α-stimulated HASMCs. Consistent with the in vitro observations, CWE inhibited the aortic expression of ICAM-1 and VCAM-1 in atherogenic diet-fed mice. CWE also downregulated the expression of nuclear factor-κB (NF-κB p65) and its uclear translocation in the stimulated HASMCs. In order to identify the active components in CWE, we re-extracted CWE using several solvents, and found that the ethyl acetate fraction was the most effective in suppressing the expression of VCAM-1 and ICAM-1. Four major acetophenones were purified from the ethyl acetate fraction, and two components, p-hydroxyacetophenone and cynandione A, potently inhibited the expression of ICAM-1 and VCAM-1 in the stimulated HASMCs. We assessed and determined the amounts of these two active components from CWE, and our results suggested that the root of C. wilfordii and its two bioactive acetophenones may be used for the prevention and treatment of atherosclerosis and vascular inflammatory diseases.

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Effects of the 4 major acetophenones, p-hydroxyacetophenone (p-HA), 2,4-dihydroxyacetophenone (2,4-DHA), cynandione A (Cyn A) and 2,5-dihydroxyacetophenone (2,5-DHA), from the EtOAc fraction of Cynanchum wilfordii extract (CWE) on the expression of cell adhesion molecules in tumor necrosis factor (TNF)-α-stimulated human aortic smooth muscle cells (HASMCs). (A) Cells were pre-treated with 10 and 50 μg/ml of each acetophenone for 2 h and then stimulated with TNF-α (10 ng/ml) for 12 h. The mRNA expression levels of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule (VCAM)-1 were determined by RT-PCR. (B) Densitometric analysis of RT-PCR is represented as the mean band density normalized to GAPDH. Results are the means ± SEM (n=3). Significantly different values are represented by symbols (**P<0.01 compared to untreated control, ##P<0.01 compared to treatment with TNF-α alone).
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f9-ijmm-35-04-0915: Effects of the 4 major acetophenones, p-hydroxyacetophenone (p-HA), 2,4-dihydroxyacetophenone (2,4-DHA), cynandione A (Cyn A) and 2,5-dihydroxyacetophenone (2,5-DHA), from the EtOAc fraction of Cynanchum wilfordii extract (CWE) on the expression of cell adhesion molecules in tumor necrosis factor (TNF)-α-stimulated human aortic smooth muscle cells (HASMCs). (A) Cells were pre-treated with 10 and 50 μg/ml of each acetophenone for 2 h and then stimulated with TNF-α (10 ng/ml) for 12 h. The mRNA expression levels of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule (VCAM)-1 were determined by RT-PCR. (B) Densitometric analysis of RT-PCR is represented as the mean band density normalized to GAPDH. Results are the means ± SEM (n=3). Significantly different values are represented by symbols (**P<0.01 compared to untreated control, ##P<0.01 compared to treatment with TNF-α alone).

Mentions: Next, we investigated whether 4 major acetophenones from the EtOAc fraction of CWE (p-HA, 2,4-DHA, Cyn A and 2,5-DHA) inhibit the TNF-α-induced expression of VCAM-1 and ICAM-1 in the HASMCs. Among these components, p-HA and Cyn A significantly inhibited the mRNA expression of VCAM-1 and ICAM-1 at 10 and 50 μg/ml (Fig. 9). However, treatment with 2,4-DHA and 2,5-DHA had little or no effect on the expression of VCAM-1 and ICAM-1. These 4 components did not affect cell viability at the concentrations tested (data not shown).


An ethanol root extract of Cynanchum wilfordii containing acetophenones suppresses the expression of VCAM-1 and ICAM-1 in TNF-α-stimulated human aortic smooth muscle cells through the NF-κB pathway.

Koo HJ, Sohn EH, Pyo S, Woo HG, Park DW, Ham YM, Jang SA, Park SY, Kang SC - Int. J. Mol. Med. (2015)

Effects of the 4 major acetophenones, p-hydroxyacetophenone (p-HA), 2,4-dihydroxyacetophenone (2,4-DHA), cynandione A (Cyn A) and 2,5-dihydroxyacetophenone (2,5-DHA), from the EtOAc fraction of Cynanchum wilfordii extract (CWE) on the expression of cell adhesion molecules in tumor necrosis factor (TNF)-α-stimulated human aortic smooth muscle cells (HASMCs). (A) Cells were pre-treated with 10 and 50 μg/ml of each acetophenone for 2 h and then stimulated with TNF-α (10 ng/ml) for 12 h. The mRNA expression levels of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule (VCAM)-1 were determined by RT-PCR. (B) Densitometric analysis of RT-PCR is represented as the mean band density normalized to GAPDH. Results are the means ± SEM (n=3). Significantly different values are represented by symbols (**P<0.01 compared to untreated control, ##P<0.01 compared to treatment with TNF-α alone).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4356471&req=5

f9-ijmm-35-04-0915: Effects of the 4 major acetophenones, p-hydroxyacetophenone (p-HA), 2,4-dihydroxyacetophenone (2,4-DHA), cynandione A (Cyn A) and 2,5-dihydroxyacetophenone (2,5-DHA), from the EtOAc fraction of Cynanchum wilfordii extract (CWE) on the expression of cell adhesion molecules in tumor necrosis factor (TNF)-α-stimulated human aortic smooth muscle cells (HASMCs). (A) Cells were pre-treated with 10 and 50 μg/ml of each acetophenone for 2 h and then stimulated with TNF-α (10 ng/ml) for 12 h. The mRNA expression levels of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule (VCAM)-1 were determined by RT-PCR. (B) Densitometric analysis of RT-PCR is represented as the mean band density normalized to GAPDH. Results are the means ± SEM (n=3). Significantly different values are represented by symbols (**P<0.01 compared to untreated control, ##P<0.01 compared to treatment with TNF-α alone).
Mentions: Next, we investigated whether 4 major acetophenones from the EtOAc fraction of CWE (p-HA, 2,4-DHA, Cyn A and 2,5-DHA) inhibit the TNF-α-induced expression of VCAM-1 and ICAM-1 in the HASMCs. Among these components, p-HA and Cyn A significantly inhibited the mRNA expression of VCAM-1 and ICAM-1 at 10 and 50 μg/ml (Fig. 9). However, treatment with 2,4-DHA and 2,5-DHA had little or no effect on the expression of VCAM-1 and ICAM-1. These 4 components did not affect cell viability at the concentrations tested (data not shown).

Bottom Line: In the present study, we evaluated the anti-inflammatory effects of an ethanol root extract of C. wilfordii (CWE) on tumor necrosis factor (TNF)-α-stimulated human aortic smooth muscle cells (HASMCs).In order to identify the active components in CWE, we re-extracted CWE using several solvents, and found that the ethyl acetate fraction was the most effective in suppressing the expression of VCAM-1 and ICAM-1.We assessed and determined the amounts of these two active components from CWE, and our results suggested that the root of C. wilfordii and its two bioactive acetophenones may be used for the prevention and treatment of atherosclerosis and vascular inflammatory diseases.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicinal and Industrial Crops, Korea National College of Agriculture and Fisheries, Jeonju 560-500, Republic of Korea.

ABSTRACT
The root of Cynanchum wilfordii (C. wilfordii) contains several biologically active compounds which have been used as traditional medicines in Asia. In the present study, we evaluated the anti-inflammatory effects of an ethanol root extract of C. wilfordii (CWE) on tumor necrosis factor (TNF)-α-stimulated human aortic smooth muscle cells (HASMCs). The inhibitory effects of CWE on vascular cell adhesion molecule (VCAM)-1 expression under an optimum extraction condition were examined. CWE suppressed the expression of VCAM-1 and ICAM-1 and the adhesion of THP-1 monocytes to the TNF-α-stimulated HASMCs. Consistent with the in vitro observations, CWE inhibited the aortic expression of ICAM-1 and VCAM-1 in atherogenic diet-fed mice. CWE also downregulated the expression of nuclear factor-κB (NF-κB p65) and its uclear translocation in the stimulated HASMCs. In order to identify the active components in CWE, we re-extracted CWE using several solvents, and found that the ethyl acetate fraction was the most effective in suppressing the expression of VCAM-1 and ICAM-1. Four major acetophenones were purified from the ethyl acetate fraction, and two components, p-hydroxyacetophenone and cynandione A, potently inhibited the expression of ICAM-1 and VCAM-1 in the stimulated HASMCs. We assessed and determined the amounts of these two active components from CWE, and our results suggested that the root of C. wilfordii and its two bioactive acetophenones may be used for the prevention and treatment of atherosclerosis and vascular inflammatory diseases.

Show MeSH
Related in: MedlinePlus