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Catechin stimulates osteogenesis by enhancing PP2A activity in human mesenchymal stem cells

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Abstract

Summary

Using human mesenchymal stem cells, we identified catechin from a panel of herbal ingredients and Chinese traditional compounds with the strongest osteogenic effects. Catechin increased alkaline phosphatase activity, calcium deposition, and mRNA expression of Runx2 and osteocalcin. We further clarified the signaling pathway that catechin mediated to stimulate osteogenesis.

Introduction

Human mesenchymal stem cells (hMSCs), useful as a species specific cell culture system for studying cell lineage differentiation, were examined as a tool to identify novel herbal ingredients and Chinese traditional compounds for enhancing osteogenesis.

Methods

Immortalized and primary hMSCs were induced in osteogenic induction medium in the presence of a variety of herbal ingredients and Chinese traditional compounds and osteogenic differentiation was evaluated by histochemical assays and quantitative RT-PCR.

Results

Using immortalized hMSCs, we first identified catechin, 18β-glycyrrhetinic acid, baishao, and danggui with osteogenic properties, which enhanced calcium deposition at the dose without significant cytotoxic effects. Primary hMSCs were then applied for confirming the osteogenic effects of catechin, which increased alkaline phosphatase activity, calcium deposition, and mRNA expression of Runx2 and osteocalcin. We further found the extracellular signal-regulated kinase (ERK) pathway was downregulated upon stimulation with catechin. Catechin increased the level and activity of protein phosphatases 2A (PP2A) that dephosphorylates ERK kinase (MEK) and ERK. Further, PP2A inhibitor, okadaic acid, abolished the effect of catechin-mediated inactivation of ERK and stimulation of osteogenesis. The blocking effect of okadaic acid on osteogenesis was further reversed by PD98059, a specific inhibitor of MEK. Co-immunoprecipitation revealed the association of PP2A to both MEK and ERK.

Conclusions

These studies propose catechin enhanced osteogenesis by increasing the PP2A level that inhibits the MEK and ERK signaling in hMSCs. These results prove the concept of using hMSCs as a convenient tool for rapid and consistent screening of the osteogenic herbal ingredients and traditional Chinese compounds.

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Acknowledgment

This study was supported in part by grants from Taipei Veterans General Hospital (V96E2-009); National Science Council (95-2314-B-075-047-MY3, 96-3111-B-010-003, 96-2627-B-010-009) and National Yang-Ming University, Ministry of Education. This work was assisted in part by the Devision of Experimental Surgery of the Department of Surgery, Taipei Veterans General Hospital.

Conflicts of interest

None.

Author information

Authors and Affiliations

  1. Laboratory of Pharmacokinetics, Institute of Traditional Medicine, National Yang-Ming University, 155, Sec 2, Li-Nong Street, Taipei, 112, Taiwan

    Y. J. Wei & T. H. Tsai

  2. Institute of Anatomy and Cell Biology, National Yang-Ming University, Taipei, Taiwan

    K. S. Tsai & M. C. Chang

  3. Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan

    Y. T. Lee & S. C. Hung

  4. Institute of Pharmacology, National Yang-Ming University, Taipei, Taiwan

    C. W. Chi & S. C. Hung

  5. Stem Cell Laboratory, Department of Medical Research and Education, Taipei Veterans General Hospital, 201, Sec 2, Shih-Pai Road, Taipei, 112, Taiwan

    K. S. Tsai, Y. T. Lee, C. W. Chi & S. C. Hung

  6. Department of Orthopaedics, Taipei Veterans General Hospital, Taipei, Taiwan

    M. C. Chang & S. C. Hung

  7. National Research Institute of Chinese Medicine, Taipei, Taiwan

    L. C. Lin

  8. Department of Education and Research, Taipei City Hospital, Taipei, Taiwan

    T. H. Tsai

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  1. Y. J. Wei
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Corresponding authors

Correspondence to T. H. Tsai or S. C. Hung.

Additional information

Yu-Jou Wei and Kuo-Shu Tsai have equally contributed to this article.

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Supplementary Fig. 1

Flow cytometric analysis of the surface markers in immortalized KP-hMSCs. Cells were reacted with FITC-conjugated antibodies (solid line) or isotype IgG antibodies (dotted line; PDF 128 kb)

Supplementary Fig. 2

Catechin enhances expression of osteogenic genes during differentiation. RT-PCR analysis for mRNA expression of Runx2, Col1A1, bone sialoprotein (BSP), osteopontin (OSP), and osteocalcin (Oc) at day 3 of osteogenic induction (PDF 84 kb)

Supplemetary Table 1

Summary of effects of each herbal ingredient and Chinese preparation on osteogenesis as analyzed by Alizarin Red S staining in primary MSCs. Data were shown as percentage (mean ± SD, n = 3) in comparison with the vehicle control (XLS 24 kb)

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Wei, Y.J., Tsai, K.S., Lin, L.C. et al. Catechin stimulates osteogenesis by enhancing PP2A activity in human mesenchymal stem cells. Osteoporos Int 22, 1469–1479 (2011). https://doi.org/10.1007/s00198-010-1352-9

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  • DOI: https://doi.org/10.1007/s00198-010-1352-9

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