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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 a 90% ethanol extract of Cynanchum wilfordii (CWE) on the adhesion of THP-1 cells to tumor necrosis factor (TNF)-α-stimulated human aortic smooth muscle cells (HASMCs). Confluent HASMCs were pre-treated for 2 h with CWE (2, 20 and 200 μg/ml) and then incubated with TNF-α (10 ng/ml) for 12 h. (A) Calcein AM-labeled THP-1 cells were added to the HASMC monolayer and allowed to adhere for 1 h. The adherence of labeled THP-1 cells to HASMCs was observed under a fluorescence microscope at x100 magnification. (B) Values are the means ± SEM of triplicate experiments. 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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f3-ijmm-35-04-0915: Effects of a 90% ethanol extract of Cynanchum wilfordii (CWE) on the adhesion of THP-1 cells to tumor necrosis factor (TNF)-α-stimulated human aortic smooth muscle cells (HASMCs). Confluent HASMCs were pre-treated for 2 h with CWE (2, 20 and 200 μg/ml) and then incubated with TNF-α (10 ng/ml) for 12 h. (A) Calcein AM-labeled THP-1 cells were added to the HASMC monolayer and allowed to adhere for 1 h. The adherence of labeled THP-1 cells to HASMCs was observed under a fluorescence microscope at x100 magnification. (B) Values are the means ± SEM of triplicate experiments. 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: We determined the effects of CWE on the adherence of THP-1 monocytes to TNF-α-stimulated HASMCs. The HASMCs were treated without or with various concentrations (2, 20 and 200 μg/ml) of CWE for 2 h prior to stimulation with TNF-α (10 ng/ml). Stimulation with TNF-α elicited a significant increase in the adhesion of THP-1 monocytes to the HASMCs (P<0.01). Treatment with CWE significantly inhibited the adhesion of the THP-1 monocytes to the HASMCs in a dose-dependent manner (Fig. 3).


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 a 90% ethanol extract of Cynanchum wilfordii (CWE) on the adhesion of THP-1 cells to tumor necrosis factor (TNF)-α-stimulated human aortic smooth muscle cells (HASMCs). Confluent HASMCs were pre-treated for 2 h with CWE (2, 20 and 200 μg/ml) and then incubated with TNF-α (10 ng/ml) for 12 h. (A) Calcein AM-labeled THP-1 cells were added to the HASMC monolayer and allowed to adhere for 1 h. The adherence of labeled THP-1 cells to HASMCs was observed under a fluorescence microscope at x100 magnification. (B) Values are the means ± SEM of triplicate experiments. 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

f3-ijmm-35-04-0915: Effects of a 90% ethanol extract of Cynanchum wilfordii (CWE) on the adhesion of THP-1 cells to tumor necrosis factor (TNF)-α-stimulated human aortic smooth muscle cells (HASMCs). Confluent HASMCs were pre-treated for 2 h with CWE (2, 20 and 200 μg/ml) and then incubated with TNF-α (10 ng/ml) for 12 h. (A) Calcein AM-labeled THP-1 cells were added to the HASMC monolayer and allowed to adhere for 1 h. The adherence of labeled THP-1 cells to HASMCs was observed under a fluorescence microscope at x100 magnification. (B) Values are the means ± SEM of triplicate experiments. 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: We determined the effects of CWE on the adherence of THP-1 monocytes to TNF-α-stimulated HASMCs. The HASMCs were treated without or with various concentrations (2, 20 and 200 μg/ml) of CWE for 2 h prior to stimulation with TNF-α (10 ng/ml). Stimulation with TNF-α elicited a significant increase in the adhesion of THP-1 monocytes to the HASMCs (P<0.01). Treatment with CWE significantly inhibited the adhesion of the THP-1 monocytes to the HASMCs in a dose-dependent manner (Fig. 3).

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