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Stimulative Effect of Ginsenosides Rg5:Rk1 on Murine Osteoblastic MC3T3‐E1 Cells
Author(s) -
Siddiqi Muhammad Hanif,
Siddiqi Muhammad Zubair,
Ahn Sungeun,
Kang Sera,
Kim YeonJu,
Veerappan Karpagam,
Yang DongUk,
Yang DeokChun
Publication year - 2014
Publication title -
phytotherapy research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.019
H-Index - 129
eISSN - 1099-1573
pISSN - 0951-418X
DOI - 10.1002/ptr.5146
Subject(s) - runx2 , ginseng , viability assay , chemistry , alkaline phosphatase , osteoblast , biochemistry , cell , pharmacology , in vitro , biology , medicine , enzyme , alternative medicine , pathology
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. Copyright © 2014 John Wiley & Sons, Ltd.