Effects of dihydrophaseic acid 3ʹ-O-β-D-glucopyranoside isolated from Lycii radicis cortex on osteoblast differentiation
Effects of dihydrophaseic acid 3ʹ-O-β-D-glucopyranoside isolated from Lycii radicis cortex on osteoblast differentiation
- 주제(키워드) herbal medicine , bioactive compound , dihydrophaseic acid 3ʹ-O-β-D-glucopyranoside , osteoblast , osteoclast , bone remodeling
- 발행기관 아주대학교
- 지도교수 김현주,정선용
- 발행년도 2017
- 학위수여년월 2017. 8
- 학위명 석사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000026018
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Ethanol extract of Lycii radicis cortex (LRC) prevented the loss of bone mineral density in ovariectomized mice by promoting the differentiation of osteoblast linage cells. In this study, I performed fractionation and isolation of the bioactive compound(s) responsible for the bone formation–enhancing effect of LRC extract. A known sesquiterpene glucoside, (1ʹR,3ʹS,5ʹR,8ʹS,2Z,4E)-dihydrophaseic acid 3ʹ-O-β-D-glucopyranoside (abbreviated as DPA3G), was isolated and identified as a candidate constituent. I investigated the effects of DPA3G on osteoblast and osteoclast differentiation, which play fundamental roles in bone formation and bone resorption, respectively, in bone remodeling. The DPA3G fraction treatment in mesenchymal stem cell line C3H10T1/2 and preosteoblast cell line MC3T3-E1 significantly enhanced cell proliferation and alkaline phosphatase activity in both cell lines compared to the untreated control cells. Furthermore, DPA3G significantly increased mineralized nodule formation and the mRNA expression of osteoblastogenesis markers, Alpl, Runx2, and Bglap, in MC3T3-E1 cells. The DPA3G treatment, however, did not influence osteoclast differentiation in primary-cultured monocytes of mouse bone marrow. Because osteoblastic and osteoclastic precursor cells coexist in vivo, I tested the DPA3G effects under the co-culture condition of MC3T3-E1 cells and monocytes. Remarkably, DPA3G enhanced not only osteoblast differentiation of MC3T3-El cells but also osteoclast differentiation of monocytes, indicating that DPA3G plays a role in the maintenance of the normal bone remodeling balance.
more목차
I. INTORDUCTION 1
II. MATERIALS AND METHODS 6
A. Fractionation, isolation, and structure elucidation of the bioactive component 6
B. Cell culture 10
C. Water-soluble tetrazolium salt (WST) assay in osteoblast cells 11
D. Alkaline phosphatase (ALP) activity assay in osteoblast cell 11
E. Mineralized nodule formation in osteoblast cells 12
F. Quantitative reverse-transcription PCR (qRT-PCR) 12
G. Osteoclastogenesis of primary monocytes and tartrate-resistant acid phosphatase (TRAP) activity assay and staining 14
H. Co-culture of MC3T3-E1 cells and primary monocytes 14
I. Statistical analysis 15
III. RESULTS 16
A. DPA3G was isolated and identified from the LRC extract as a bioactive component for enhancing osteoblast differentiation 16
B. DPA3G increased the cellular proliferation, differentiation, and mineralized nodule formation of osteoblasts 18
C. DPA3G did not influence differentiation of osteoclasts 24
D. DPA3G enhanced both osteoblast and osteoclast differentiation in the MC3T3-E1 and primary monocyte co-culture system 27
IV. DISCUSSION 32
V. CONCLUSIONS 34
REFERENCES 35
국문요약 43
