J Korean Soc Menopause.  2012 Aug;18(2):100-105.

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

• Kang YH
• Lee DY
• Choi DS
• Yoon BK
• Lee YJ
• Rhyu MR

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.

Keyword

Cardiovascular diseases; Cimicifuga; Estrogens; Vascular smooth muscle cell

MeSH Terms

1-Propanol
2-Propanol
Animals
Aorta
Bromodeoxyuridine
Cardiovascular Diseases
Cell Proliferation
Cimicifuga
Estrogens
Ethanol
Female
Humans
Indicators and Reagents
Menopause
Methanol
Muscle, Smooth, Vascular
Rats
Reactive Oxygen Species
Vitamin E
Vitamins
1-Propanol
2-Propanol
Bromodeoxyuridine
Estrogens
Ethanol
Indicators and Reagents
Methanol
Reactive Oxygen Species
Vitamin E
Vitamins

Figure

• 

  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.

• 

  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.

• 

  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.

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