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Kang, Lee, Choi, Yoon, Lee, and Rhyu: Effect of Korean Native Cimicifuga Species on Proliferation of Vascular Smooth Muscle Cells
Original Article
The Journal of Korean Society of Menopause

The Journal of Korean Society of Menopause 2012; 18(2): 100-105.

Published online: 29 August 2012

DOI: https://doi.org/10.6118/jksm.2012.18.2.100

Effect of Korean Native Cimicifuga Species on Proliferation of Vascular Smooth Muscle Cells

Young-Hee Kang, M.S.1, Dong-Yun Lee, M.D.2, Doo Seok Choi, M.D.1, 2, Byung-Koo Yoon, M.D.1, 2, Young Joo Lee, Ph.D.3, Mee-Ra Rhyu, Ph.D.4

1Samsung Biomedical Research Institute, Seoul, Korea.

2Department of Obstetrics and Gynecology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.

3Department of Bioscience and Biotechnology, Sejong University, Seoul, Korea.

4Divisions of Metabolism and Functionality Research, Korea Food Research Institute, Seongnam, Korea.

Address for Correspondence: Byung-Koo Yoon, Department of Obstetrics and Gynecology, Samsung Medical Center, Sungkyunkwan University School of Medicine, 50 Irwon-dong, Gangnam-gu, Seoul 135-710, Korea. Tel: +82-2-3410-3519, Fax: +82-2-3410-0630, bkyoon@skku.edu

Received 4 November 2011       Revised 1 December 2011       Accepted 1 December 2011

Copyright © 2012 by The Korean Society of Menopause

(open-access):

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/).

Abstract

Objectives

To examine the effect of extracts of Korean native Cimicifuge species on cell proliferation in vascular smooth muscle cells (VSMC).

Methods

VSMC were isolated from rat aorta. Cell proliferation was assessed by measure of bromodeoxyuridine incorporation into the cells. Differences in Reactive oxygen species (ROS) levels were examined after exposure to the extracts of Korean native Cimicifuge species using the detection reagents dichlorofluorecin diacetate. The rhizomes/roots were air-dried and milled with a commercial food mixer. Milled rhizomes/roots of each Cimicifuga species were separately extracted by 80% ethanol, absolute methanol, and 40% 2-propanol using homogenizer and evaporated under reduced pressure at low temperatures. Effects of extracts dissolved in phosphate-buffered saline (0.3 mg/mL) were examined.

Results

Ethanolic, methanolic or propanolic extracts of 4 Korean native Cimicifuge species (Cimicifuga [C] davurica, C. japonica, C. heracleifolia var. bifida Nakai, C. simplex) were screened. The addition of extracts of each Korean native Cimicifuge species to cells in the presence of 10% fetal bovine serum (FBS) potently inhibited cell proliferation. Significant decrease of 23%-30% was observed. Vitamin E, a potent antioxidant, inhibited 10% FBS-stimulated cell proliferation of VSMC. We also demonstrated that extracts of each Korean native Cimicifuge species decreased intracellular ROS generation induced with 10% FBS. The effect of Korean native Cimicifuge species was not species-specific and solvent-specific.

Conclusion

TExtracts of Korean native Cimicifuge species inhibit VSMC proliferation via inhibition of intracellular ROS. These findings suggest that Cimicifuge species used for reducing menopause symptoms might be cardioprotective in women.

Keywords: Cardiovascular diseases, Cimicifuga, Estrogens, Vascular smooth muscle cell

Figures and Tables

Fig. 1
The effects of extracts of Cimicifuga [C] species (0.3 mg/mL) on 10% fetal bovine serum (FBS)-stimulated proliferation of vascular smooth muscle cells. The extracts were added to the medium containing 10% FBS and the incubation proceeded for 24 h. Cell proliferation was measured by bromodeoxyuridine assay. CJ: C. japonica, CH: C. heracleifolia var. bifida Nakai, CD: C. davurica, CS: C. simplex, DSF: defined serum-free medium, P: 40% 2-propanol, M: absolute methanol, E: 80% ethanol. *P < 0.05 vs. control.
jksm-18-100-g001
Fig. 2
The effect of vitamin E on 10% fetal bovine serum (FBS)-stimulated proliferation of vascular smooth muscle cells, as assessed by bromodeoxyuridine assay. Vitamin E (40 µM) was added to the medium containing 10% FBS and the incubation proceeded for 24 h. DSF: defined serum-free medium. *P < 0.05 vs. DSF; **P < 0.05 vs. 10% FBS.
jksm-18-100-g002
Fig. 3
The effects of extracts of Cimicifuge species (0.3 mg/mL) on reactive oxygen species (ROS) generation of vascular smooth muscle cells, as assessed by fluorescence-activated cell sorter (FACs) analysis. The extracts were added to the medium containing 10% fetal bovine serum (FBS). P: 40% 2-propanol, M: absolute methanol, E: 80% ethanol. *P < 0.05 vs. control. CJ: C. japonica, CH: C. heracleifolia var. bifi da Nakai, CD: C. davurica, CS: C. simplex.
jksm-18-100-g003

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