Vitisin B stimulates osteoblastogenesis via estrogen receptor-mediated activation of non-genomic Src and MAPKs pathway

Background/Purpose: Vitisin B is a major component existed in Vitis thunbergii, a herbal medicine used in Taiwan for treatment of inflammatory bone diseases. We recently reported that vitisin B stimulated differentiation in primary cultured osteoblasts and treatment with vitisin B-enriched preparation obviously ameliorated ovariectomy-induced bone loss in mice. This study further delineated the action mechanism(s) that how vitisin B stimulates osteoblastogenesis by using MC3T3-E1 osteoblasts. Methods: Cell differentiation and mineralization were identified by alkaline phosphatase (ALP) activity and Alizarin red S staining, respectively. RT-PCR and western blot were used to analyze the expression of osteoblast-associated genes and signal molecules. The transcriptional activity of estrogen receptor (ER) was also assessed. Results: Vitisin B significantly increased ALP activity, bone mineralization, mRNA expression of osteoids (type 1 collagen, bone sialprotein and osteocalcin) and bone-characteristic transcription factors (runt-related transcription factor-2 and osterix) through ER since such responsiveness were obviously repressed by ER antagonist ICI182,780. Unlike 17β-estradiol (E2), vitisin B failed to stimulate either ERαor ERβmediated transcriptional activity. Nevertheless, vitisin B rapidly induced ERα and Src phosphorylations within 5 min and evoked late phosphorylations of p38 and ERK after 15-30 min stimulation through ER. Furthermore, p38 inhibitor SB203580 and MEK inhibitor PD98059 significantly inhibited vitisin B induced differentiation. Src inhibitor PP2 significantly repressed vitisin B-induced activation of MAPK and final mineralization suggesting that Src might play an important role. Conclusions: Vitisin B

[1]  Yu-Ling Huang,et al.  A Special Ingredient (VtR) Containing Oligostilbenes Isolated from Vitis thunbergii Prevents Bone Loss in Ovariectomized Mice: In Vitro and In Vivo Study , 2013, Evidence-based complementary and alternative medicine : eCAM.

[2]  Lie-Chwen Lin,et al.  Neobavaisoflavone stimulates osteogenesis via p38-mediated up-regulation of transcription factors and osteoid genes expression in MC3T3-E1 cells. , 2012, Phytomedicine : international journal of phytotherapy and phytopharmacology.

[3]  R. Choi,et al.  Baicalin, a flavone, induces the differentiation of cultured osteoblasts: an action via the Wnt/β-catenin signaling pathway. , 2012, The Journal of Biological Chemistry.

[4]  Yu-Ling Huang,et al.  Ugonin K-stimulated osteogenesis involves estrogen receptor-dependent activation of non-classical Src signaling pathway and classical pathway. , 2012, European journal of pharmacology.

[5]  Yu-Ling Huang,et al.  Ugonin K promotes osteoblastic differentiation and mineralization by activation of p38 MAPK- and ERK-mediated expression of Runx2 and osterix. , 2011, European journal of pharmacology.

[6]  W. Chiou,et al.  8-Prenylkaempferol accelerates osteoblast maturation through bone morphogenetic protein-2/p38 pathway to activate Runx2 transcription. , 2011, Life sciences.

[7]  Michael D. Schneider,et al.  The p38 MAPK pathway is essential for skeletogenesis and bone homeostasis in mice. , 2010, The Journal of clinical investigation.

[8]  L. Yu-Lee,et al.  Src family kinase/abl inhibitor dasatinib suppresses proliferation and enhances differentiation of osteoblasts , 2010, Oncogene.

[9]  M. Piccart,et al.  The Src inhibitor dasatinib accelerates the differentiation of human bone marrow-derived mesenchymal stromal cells into osteoblasts , 2010, BMC Cancer.

[10]  M. Rybchyn,et al.  Osteoblasts play key roles in the mechanisms of action of strontium ranelate , 2009, British journal of pharmacology.

[11]  Yu-Ling Huang,et al.  Oligostilbenes from the roots of Vitis thunbergii. , 2009, Planta medica.

[12]  Baojie Li,et al.  Protein Palmitoylation Regulates Osteoblast Differentiation through BMP-Induced Osterix Expression , 2009, PloS one.

[13]  W. Fu,et al.  Enhancement of bone morphogenetic protein-2 expression and bone formation by coumarin derivatives via p38 and ERK-dependent pathway in osteoblasts. , 2008, European journal of pharmacology.

[14]  Hong-hao Zhou,et al.  Genistein stimulates osteoblastic differentiation via p38 MAPK-Cbfa1 pathway in bone marrow culture , 2007, Acta Pharmacologica Sinica.

[15]  M. Yen,et al.  Forkhead Proteins Are Critical for Bone Morphogenetic Protein-2 Regulation and Anti-tumor Activity of Resveratrol* , 2007, Journal of Biological Chemistry.

[16]  Renny T. Franceschi,et al.  Critical role of the extracellular signal–regulated kinase–MAPK pathway in osteoblast differentiation and skeletal development , 2007, The Journal of cell biology.

[17]  Wei Fang Chen,et al.  Diosgenin Induces Hypoxia-Inducible Factor-1 Activation and Angiogenesis through Estrogen Receptor-Related Phosphatidylinositol 3-kinase/Akt and p38 Mitogen-Activated Protein Kinase Pathways in Osteoblasts , 2005, Molecular Pharmacology.

[18]  E. Levin,et al.  Integration of the extranuclear and nuclear actions of estrogen. , 2005, Molecular endocrinology.

[19]  Yu-Ling Huang,et al.  Resveratrol derivatives from the roots of Vitis thunbergii. , 2005, Journal of natural products.

[20]  R. Roskoski,et al.  Src protein-tyrosine kinase structure and regulation. , 2004, Biochemical and biophysical research communications.

[21]  Frank Claessens,et al.  Selective DNA recognition by the androgen receptor as a mechanism for hormone-specific regulation of gene expression. , 2003, Molecular genetics and metabolism.

[22]  A. Bilancio,et al.  Src Is an Initial Target of Sex Steroid Hormone Action , 2002, Annals of the New York Academy of Sciences.

[23]  S. Mohan,et al.  Two Different Pathways for the Maintenance of Trabecular Bone in Adult Male Mice , 2002, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[24]  E. Levin.  highlighted topics Genome and Hormones: Gender Differences in Physiology Invited Review: Cell localization, physiology, and nongenomic actions of estrogen receptors , 2001 .

[25]  S. Safe,et al.  Estrogen Regulation of Cyclin D1 Gene Expression in ZR-75 Breast Cancer Cells Involves Multiple Enhancer Elements* , 2001, The Journal of Biological Chemistry.

[26]  B. Katzenellenbogen,et al.  Nongenotropic, Sex-Nonspecific Signaling through the Estrogen or Androgen Receptors Dissociation from Transcriptional Activity , 2001, Cell.

[27]  J. Bonjour,et al.  Printed in U.S.A. Copyright © 1999 by The Endocrine Society Regulation of Alkaline Phosphatase Activity by p38 MAP Kinase in Response to Activation of Gi Protein-Coupled Receptors by Epinephrine in Osteoblast-Like Cells* , 2022 .

[28]  K. Korach,et al.  Estrogen receptor gene disruption: molecular characterization and experimental and clinical phenotypes. , 1996, Recent progress in hormone research.