title: Stimulative effect of ginsenosides Rg5:Rk1 on murine osteoblastic MC3T3-E1 cells.
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.
authors.affiliation: Ginseng Genetic Resource Bank, Graduate School of Biotechnology, College of Life Science, Kyung Hee University, Suwon, 449-701, Korea.
authors.forename: Muhammad Hanif
authors.initials: MH
authors.lastname: Siddiqi
authors.forename: Muhammad Zubair
authors.initials: MZ
authors.lastname: Siddiqi
authors.forename: Sungeun
authors.initials: S
authors.lastname: Ahn
authors.forename: Sera
authors.initials: S
authors.lastname: Kang
authors.forename: Yeon-Ju
authors.initials: YJ
authors.lastname: Kim
authors.forename: Karpagam
authors.initials: K
authors.lastname: Veerappan
authors.forename: Dong-Uk
authors.initials: DU
authors.lastname: Yang
authors.forename: Deok-Chun
authors.initials: DC
authors.lastname: Yang
mesh_terms: D016475:3T3 Cells; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival; D024042:Collagen Type I / Q000378:metabolism; D050659:Core Binding Factor Alpha 1 Subunit / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D036145:Ginsenosides / Q000494:pharmacology*; D051379:Mice; D010006:Osteoblasts / Q000187:drug effects*; D005894:Panax / Q000737:chemistry
chemical_list: C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2; D024042:Collagen Type I; D050659:Core Binding Factor Alpha 1 Subunit; D036145:Ginsenosides; C493716:Runx2 protein, mouse; C572381:ginsenoside Rg5; D000469:Alkaline Phosphatase
keywords: Panax ginseng; alkaline phosphatase; bone formation; ginsenosides; mineralization; osteoblast