Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (19): 3523-3526.doi: 10.3969/j.issn.1673-8225.2010.19.021

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Expression of osteogenesis- and adipogenesis-related transcription factors during osteogenic differentiation of rat mesenchymal stem cells induced by eucommia bark

Zhang Xian1, Cai Jian-ping1, Zhang Yan-hong2, Tan Xiang-ling2   

  1. 1Department of Orthopaedics, Wuxi Hospital of Traditional Chinese Medicine, Wuxi  214001, Jiangsu Province, China;
    2Institute of Bioengineering, Nantong University, Nantong  226007, Jiangsu Province, China
  • Online:2010-05-07 Published:2010-05-07
  • Contact: Tan Xiang-ling, Professor, Institute of Bioengineering, Nantong University, Nantong 226007, Jiangsu Province, China tanxl@ntu.edu.cn
  • About author:Zhang Xian, Associate chief physician, Department of Orthopaedics, Wuxi Hospital of Traditional Chinese Medicine, Wuxi 214001, Jiangsu Province, China zhangxian0772@sina.com
  • Supported by:

    the Social Development Program of Bureau of Science and Technology of Wuxi City of Jiangsu Province in 2007, No. CSE00713*

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Abstract:

BACKGROUND: Eucommia bark has obvious effects of intervention and treatment on osteoporosis. Previous studies have confirmed that eucommia bark can significantly induce differentiation of bone marrow mesenchymal stem cells (BMSCs) into osteoblasts and adipogenic cells. There are no reports concerning changes in osteogenesis- and adipogenesis-related transcription factor expression.

OBJECTIVE: To observe changes in expression of osteogenic and adipogenic differentiation related transcription factors during osteogenic differentiation of rat BMSCs induced by eucommia bark.

METHODS: Water and MeOH extract of eucommia bark was prepared. BMSCs from Sprague Dawley rat were cultured in vitro for 3 passages. Extract was volume metered into 15 mL as stock solution according to 2 g of crude drug. Extract of eucommia bark were diluted to 10-3 and 10-4 groups. The test contained six groups. Negative control group received the same volume of phosphate buffered saline as inducers. Positive control group induced by dexamethasone 0.01 mmol/L, β-sodium glycerophosphate 10 mmol/L and L-sodium ascorbate 50 mg/L. There were water extract of eucommia bark 10-4 dilution group, water extract of eucommia bark 10-3 dilution group, MeOH extract of eucommia bark 10-4 dilution group and MeOH extract of eucommia bark 10-3 dilution group. The culture was stopped at 15 days following induction in each group. Reverse transcription PCR and fluorescent quantitative PCR were used to detect expression of osteogenic transcription factor Runx2 and Osterix, and adipogenic transcription factor peroxisome proliferator activated receptor γ (PPARg) and fatty acid binding protein.

RESULTS AND CONCLUSION: The expression of Runx2 did not change except downregulated at 10-3 dilution of water extract and upregulated at 10-4 dilution of MeOH extract. Osx was downregulated obviously in all induction groups, and the expression of PPARr did not change and fatty acid binding protein was downregulated in all induction groups remarkably. These results suggested that of MSCs induced by eucommia bark was related to inhibition of the expression of fatty acid binding protein.

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