Stimulative effect of ginsenosides Rg5:Rk1 on murine osteoblastic MC3T3-E1 cells

Phytother Res. 2014 Oct;28(10):1447-55. doi: 10.1002/ptr.5146. Epub 2014 Mar 19.

Abstract

Panax ginseng C.A. Meyer (P. ginseng), hereafter referred to as P. ginseng, is known to exert a wide range of pharmacological effects both in vitro and in vivo; however, few studies have investigated the effects of ginseng on bone metabolism. We therefore investigated the potential antiosteoporotic properties of ginseng on the growth and differentiation of murine MC3T3-E1 cells. Rg5:Rk1 is a mixture of protopanaxadiol-type ginsenosides, isolated from fresh P. ginseng root, via a repetitive steaming and drying process. In this study, we examined the stimulatory effects of Rg5:Rk1 on the differentiation and mineralization of MC3T3-E1 cells. Undifferentiated cells were treated with a range of concentrations of Rg5:Rk1 (1-50 µg/mL), and cell viability was measured with the 3-(4,5-dimethyl-thiazol-2yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Treatment with Rg5:Rk1 significantly increased cell viability in a dose-dependent manner. To investigate the possible mechanisms by which Rg5:Rk1 affects the early differentiation phase of MC3T3-E1 cells, the cells were treated with Rg5:Rk1 for 14-24 days before assessing the levels of multiple osteoblastic markers. The markers examined included alkaline phosphatase (ALP) activity type I collagen content (Coll-I), calcium deposition (by Alizarin Red S staining), extracellular mRNA expression of bone morphogenetic protein-2 (BMP-2), and the level of Runt-related transcription factor 2 (Runx2). Rg5:Rk1 treatment also increased the activities of proteins associated with osteoblast growth and differentiation in a dose-dependent manner. Overall, we found that the Rg5:Rk1 mixture of ginsenosides improved the osteoblastic function of MC3T3-E1 cells by increasing their proliferative capacity. This improvement is due to the action of Rg5:Rk1 on BMP-2, which is mediated by Runx2-dependent pathways.

Keywords: Panax ginseng; alkaline phosphatase; bone formation; ginsenosides; mineralization; osteoblast.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Alkaline Phosphatase / metabolism
  • Animals
  • Bone Morphogenetic Protein 2 / metabolism
  • Cell Differentiation / drug effects
  • Cell Proliferation / drug effects
  • Cell Survival
  • Collagen Type I / metabolism
  • Core Binding Factor Alpha 1 Subunit / metabolism
  • Dose-Response Relationship, Drug
  • Ginsenosides / pharmacology*
  • Mice
  • Osteoblasts / drug effects*
  • Panax / chemistry

Substances

  • Bmp2 protein, mouse
  • Bone Morphogenetic Protein 2
  • Collagen Type I
  • Core Binding Factor Alpha 1 Subunit
  • Ginsenosides
  • Runx2 protein, mouse
  • ginsenoside Rg5
  • Alkaline Phosphatase