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Anti-osteoclastogenic effect and action mechanism of the sargachromanol G isolated from Sargassum siliquastrum

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Alternative Title
Sargassum siliquastrum로부터 분리한 sargachromanol G의 파골세포형성 억제 효과 및 작용 기전
Abstract
Inflammatory bone diseases are characterized by the presence of pro-inflammatory cytokines that regulate bone turnover. The receptor activator of NF-κB ligand (RANKL) is a soluble osteoblast-derived protein that induces bone resorption through osteoclast differentiation and activation. Sargachromanol G is isolated from Sargassum siliquastrum and has cytotoxicity, antioxidant, and antiviral activities. The active components and underlying mechanisms of its anti-osteoclastogenic activity remain largely unknown. In the present study, we investigated the anti-osteoclastogenic effects of sargachromanol G isolated from S. siliquastrum on the expression of IL-1β-induced osteoclastogenic factors (RANKL, IL-6, PGE2 and COX-2) in human osteoblast MG-63 cells, as well as LPS or RANKL-induced pro-inflammatory factors and osteoclastogenic factors (nitric oxide (NO), cytokines (TNF-α, IL-1β and IL-6), TRAP, CTR, TRAF6, Cath-K, and MMP-9) in murine macrophage RAW 264.7 cells. We also examined the role of nuclear factor-κB (NF-κB) and mitogen activated protein kinase (MAPK) signaling induced by IL-1β in MG-63 and LPS or RANKL in RAW 264.7 cells. Sargachromanol G dose-dependently inhibited the production of osteoclastogenic factors in MG-63 and RAW 264.7 cells. Sargachromanol G also inhibited phosphorylation of NF-κB (IκB-α, p65 and p50) and MAPK (ERK1/2, JNK and p38). These results suggest that the anti-osteoclastogenic activity of sargachromanol G isolated from S. siliquastrum may result from modulation of osteoclastogenic factors and cytokines via suppression of phospholrylated MAPK and NF-κB activation.
Author(s)
Yoon, Weon Jong
Issued Date
2010
Awarded Date
2010. 8
Type
Dissertation
URI
http://dcoll.jejunu.ac.kr/jsp/common/DcLoOrgPer.jsp?sItemId=000000005151
Alternative Author(s)
윤원종
Affiliation
제주대학교 일반대학원
Department
대학원 의학과
Advisor
유은숙
Table Of Contents
CONTENTS
ABSTRACT ..............................Ⅰ
CONTENTS ..............................Ⅱ
LIST OF SCHEME ...........................Ⅴ
LIST OF TABLES ...........................Ⅵ
LIST OF FIGURES ...........................Ⅶ

1. Introduction .............................1
2. Material and Methods ........................7
2-1. Chemicals and reagents
2-2. Isolation of sargachromanol G
2-3. Cell culture
2-4. Cell viability
2-5. Measurement of PGE2 production
2-6. Measurement of cytokines (TNF-α, IL-1β and IL-6) production
2-7. Measurement of NO production
2-8. Immunoblotting analysis
2-9. RT-PCR analysis
2-10. Tartrate-resistant acid phosphatase (TRAP) staining and assay
2-11. Statistical analysis
3. Results
3-1. Results
Isolation of sargachromanol G and effect of sagachromanol G in IL-1β-stimulated osteoblast
3-1-1. Sargachromanol G isolated from Sargassum siliquastrum
3-1-2. Effects of solvent fractions and sargachromanol G from S. siliquastrum on the cell viability in MG-63 cells
3-1-3. Effects of solvent fractions and sargachromanol G on PGE2 production in IL-1β-stimulated MG-63 cells
3-1-4. Effects of solvent fractions and sargachromanol G on IL-6 production in IL-1β-stimulated MG-63 cells
3-1-5. Effects of sargachromanol G on protein levels of COX-2, RANKL and OPG in IL-1β-stimulated MG-63 cells
3-1-6. Effect of sargachromanol G on NF-κB signaling pathway in IL-1β-stimulated MG-63 cells
3-1-7. Effects of sargachromanol G on MAPKs signaling pathway in IL-1β-stimulated MG-63 cells
3-2. Results
Effect of sargachromanol G in LPS-stimulated macrophage
3-2-1. Effects of solvent fractions and sargachromanol G from S. siliquastrum on the cell viability in RAW 264.7 cells
3-2-2. Effects of solvent fractions and sargachromanol G from S. siliquastrum on NO production and cytotoxicity in LPS-stimulated RAW 264.7 cells
3-2-3. Effects of solvent fractions and sargachromanol G from S. siliquastrum on PGE2 production in LPS-stimulated RAW 264.7 cells
3-2-4. Effects of sargachromanol G on protein levels of iNOS and COX-2 in LPS-stimulated RAW 264.7 cells
3-2-5. Effects of sargachromanol G on pro-inflammatory cytokines production in LPS-stimulated RAW 264.7 cells
3-2-6. Effect of sargachromanol G on NF-κB signaling pathway in LPS-stimulated RAW 264.7 cells
3-2-7. Effects of sargachromanol G on MAPKs signaling pathway in LPS-stimulated RAW 264.7 cells
3-3. Results
Effect of sargachromanol G in RANKL- stimulated preosteoclast
3-3-1. Effects of sargachromanol G on osteoclastogenic factors in RANKL-stimulated RAW 264.7 cells
3-3-2. Effect of sargachromanol G on osteoclast differentiation from RANKL-stimulated RAW 264.7 cells
3-3-3. Effect of sargachromanol G on NF-κB signaling pathway in RANKL-stimulated RAW 264.7 cells
3-3-4. Effects of sargachromanol G on MAPKs signaling pathway in RANKL-stimulated RAW 264.7 cells
3-3-5. Effects of sargachromanol G on transcription factors related-osteoclastogenesis in RANKL-stimulated RAW 264.7 cells
4. Discussion ..............................86
5. Reference ..............................94
Degree
Doctor
Publisher
제주대학교 일반대학원
Citation
Yoon, Weon Jong. (2010). Anti-osteoclastogenic effect and action mechanism of the sargachromanol G isolated from Sargassum siliquastrum
Appears in Collections:
General Graduate School > Medicine
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