Korean J Physiol Pharmacol.  2014 Aug;18(4):347-352. 10.4196/kjpp.2014.18.4.347.

Effects of Watercress Containing Rutin and Rutin Alone on the Proliferation and Osteogenic Differentiation of Human Osteoblast-like MG-63 Cells

• Hyun H
• Park H
• Jeong J
• Kim J
• Kim H
• Oh HI
• Hwang HS
• Kim HH

• KMID: 2285530
• DOI: http://doi.org/10.4196/kjpp.2014.18.4.347

Abstract

Most known osteoporosis medicines are effective for bone resorption, and so there is an increasing demand for medicines that stimulate bone formation. Watercress (N. officinale R. Br.) is widely used as a salad green and herbal remedy. This study analyzed a watercress extract using ultra-performance liquid chromatography/mass spectrometry, and identified a rutin as one of its major constituents. Osteogenic-related assays were used to compare the effects of watercress containing rutin (WCR) and rutin alone on the proliferation and differentiation of human osteoblast-like MG-63 cells. The reported data are expressed as percentages relative to the control value (medium alone; assigned as 100%). WCR increased cell proliferation to 125.0+/-4.0% (mean+/-SD), as assessed using a cell viability assay, and increased the activity of alkaline phosphatase, an early differentiation marker, to 222.3+/-33.8%. In addition, WCR increased the expression of collagen type I, another early differentiation marker, to 149.2+/-2.8%, and increased the degree of mineralization, a marker of the late process of differentiation, to 122.9+/-3.9%. Rutin alone also increased the activity of ALP (to 154.4+/-12.2%), the expression of collagen type I (to 126.6+/-6.2%), and the degree of mineralization (to 112.3+/-5.0%). Daidzein, which is reported to stimulate bone formation, was used as a positive control; the effects of WCR on proliferation and differentiation were significantly greater than those of daidzein. These results indicate that WCR and rutin can both induce bone formation via the differentiation of MG-63 cells. This is the first study demonstrating the effectiveness of either WCR or rutin as an osteoblast stimulant.

Keyword

Differentiation; Osteoblast; Proliferation; Rutin; Watercress

MeSH Terms

Alkaline Phosphatase
Bone Resorption
Cell Proliferation
Cell Survival
Collagen Type I
Humans
Osteoblasts
Osteogenesis
Osteoporosis
Rutin*
Spectrum Analysis
Alkaline Phosphatase
Collagen Type I
Rutin

Figure

• 

  Fig. 1 PDA chromatogram (upper) of MS positive ion electrospray (lower) analyses of (A) WCR and (B) rutin.

• 

  Fig. 2 Effects of WCR and rutin at various concentrations (µg/ml) on cell proliferation in MG-63 cells. The data are expressed as percentages relative to the control value (assigned a value of 100%), and are mean and SD values of three independent samples. *p<0.05, **p<0.01, significantly different from the control group.

• 

  Fig. 3 Stimulation of ALP activity in MG-63 cells by WCR and rutin at various concentrations (µg/ml). The data are expressed as percentages relative to the control value (assigned a value of 100%), and are mean and SD values of three independent samples. *p<0.05, **p<0.01, ***p<0.001, significantly different from the control group.

• 

  Fig. 4 Effects of WCR and rutin at various concentrations (µg/ml) on the collagen content of MG-63 cells after 5 days. The data are percentages relative to the control value (assigned a value of 100%), and are mean and SD values of three independent samples. **p<0.01, ***p<0.001, significantly different from the control group.

• 

  Fig. 5 Effects of WCR and rutin at various concentrations (µg/ml) on mineralization of the extracellular matrix in MG-63 cells after 7 days. The data are expressed as percentages relative to the control value (assigned a value of 100%), and are mean and SD values of three independent samples. ARS, Alizarin red staining. *p<0.05, **p<0.01, ***p<0.001, significantly different from the control group.

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