| Project/Area Number |
14580618
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Bioorganic chemistry
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| Research Institution | TOKYO UNIVERSITY OF SCIENCE |
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Principal Investigator |
HAKAMATSUKA Takashi Tokyo University of Science, Faculty of Pharmaceutical Sciences, Associate Professor, 薬学部, 助教授 (60221488)
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| Co-Investigator(Kenkyū-buntansha) |
KINOSHITA Takeshi Teikyo University, Faculty of Pharmaceutical Sciences, Associate Professor, 薬学部, 助教授 (10107386)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2004: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2003: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2002: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | bone / bone remodeling / natural products / gene expression / Saos-2 / OCIF / 骨芽細胞 / 破骨細胞 / 南西諸島 / 薬用植物 / フラボノイド / 沖縄 / 遺伝子 / モノテルペン |
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| Research Abstract |
We have established a PCR-based bioassay to search for biologically active compounds that can modulate the expression of genes encoding bone remodeling factors. Changes in the level of target mRNA in cell cultures were quantitatively determined by competitive PCR using internal standard DNA. With this bioassay system, we have been searching for lipophilic low-molecular-weight probes that can easily pass through the cell membrane and have an effect on some signal transduction steps. The compounds discovered will be useful tools to elucidate the mechanisms of intracellular signalings guiding the expression of the target genes.
As an application of the screening strategy, the expression of the PEBP2αA/CBFa1 and OPG/OCIF genes in MC3T3-E1 cells, a mouse osteoblast-like cell line, was examined. Screening of Ryukyu plants demonstrated that the methanol extracts of Oreocnide pedunculata branches and Wikstroemia retusa leaves enhanced the expression of the PEBP2αA/CBFa1 gene and OPG/OCIF gene, respectively. Furthermore, the methanol extracts of Ryukyu plants were assessed for the capacity to enhance the gene expression of interleukin-2 in Jurkat cells, a human T cell line. Chromatography by activity-guided fractionation yielded the active principles of Cerbera manghas branches and Raphiolepis indica branches and they were identified by spectroscopic examination as monoterpene cerbinal and flavonoid epicatechin, respectively.
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