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      Multiple signaling pathways involved in stimulation of osteoblast differentiation by N-methyl- D-aspartate receptors activation in vitro

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          Abstract

          Aim:

          Glutamate receptors are expressed in osteoblastic cells. The present study was undertaken to investigate the mechanisms underlying the stimulation of osteoblast differentiation by N-methyl- D-aspartate (NMDA) receptor activation in vitro.

          Methods:

          Primary culture of osteoblasts was prepared from SD rats. Microarray was used to detect the changes of gene expression. The effect of NMDA receptor agonist or antagonist on individual gene was examined using RT-PCR. The activity of alkaloid phosphotase (ALP) was assessed using a commercial ALP staining kit.

          Results:

          Microarray analyses revealed that 10 genes were up-regulated by NMDA (0.5 mmol/L) and down-regulated by MK801 (100 μmol/L), while 13 genes down-regulated by NMDA (0.5 mmol/L) and up-regulated by MK801 (100 μmol/L). Pretreatment of osteoblasts with the specific PKC inhibitor Calphostin C (0.05 μmol/L), the PKA inhibitor H-89 (20 nmol/L), or the PI3K inhibitor wortmannin (100 nmol/L) blocked the ALP activity increase caused by NMDA (0.5 mmol/L). Furthermore, NMDA (0.5 mmol/L) rapidly increased PI3K phosphorylation, which could be blocked by pretreatment of wortmannin (100 nmol/L).

          Conclusion:

          The results suggest that activation of NMDA receptors stimulates osteoblasts differentiation through PKA, PKC, and PI3K signaling pathways, which is a new role for glutamate in regulating bone remodeling.

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          Most cited references 59

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          The regulation of the transcription factor NF-kappaB activity occurs at several levels including controlled cytoplasmic-nuclear shuttling and modulation of its transcriptional activity. A critical component in NF-kappaB regulation is the IkappaB kinase (IKK) complex. This review is focused on recent progress as well as unanswered questions regarding the regulation and function of NF-kappaB and IKK.
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                Author and article information

                Journal
                Acta Pharmacol Sin
                Acta Pharmacol. Sin
                Acta Pharmacologica Sinica
                Nature Publishing Group
                1671-4083
                1745-7254
                July 2011
                20 June 2011
                : 32
                : 7
                : 895-903
                Affiliations
                [1 ]Department of Endocrine and Metabolic Diseases, Shanghai Institute of Endocrine and Metabolic Diseases, Shanghai Clinical Center for Endocrine and Metabolic Diseases, Ruijin Hospital, Shanghai Jiaotong University School of Medicine , Shanghai 200025, China;
                [2 ]Laboratory of Endocrine & Metabolic Diseases, Institute of Health Science, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences , Shanghai 200025, China;
                [3 ]Shanghai Institute of Traumatology and Orthopaedics, Department of Orthopaedics, Ruijin Hospital, Shanghai Jiaotong University School of Medicine , Shanghai 200025, China
                Author notes
                Article
                aps201138
                10.1038/aps.2011.38
                4003127
                21685927
                Copyright © 2011 CPS and SIMM
                Categories
                Original Article

                Pharmacology & Pharmaceutical medicine

                bone, osteoblast, glutamate receptor, nmda receptor, pka, pkc, pi3k

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