지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
커뮤니티
연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학위논문
- 저자정보
- 지도교수
- 손영주, 정혁상
- 발행연도
- 2017
- 저작권
- 경희대학교 논문은 저작권에 의해 보호받습니다.
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Background and Purposes : Osteoporosis is a condition
characterized by low bone mass and increased bone fragility. It
has become a major problem of senior citizens. Recently, many
studies of the bone remodelling process suggest that factors
involved in inflammation are linked with osteoporosis. The
purpose of this study is to experiment the effect of water extract
of Forsythiae Fructus (wFF) on osteoclast differentiation; and the
other purpose is to examine the effect of wFF on osteoporosis in
ovariectomized rat.
Materials and methods : To test the effect of wFF on
osteoclast differentiation and suppression of gene expression
related to bone resorption RAW 264.7 cells were used. Various
level of concentration of wFF (50, 100, 200 μg/mL) were added to
RAW 264.7 cells co-cultured with RANKL. The number of TRAP
positive cell, TRAP activity, Pit area, mRNA expression of makers
(RANK, TRAP, CAⅡ, CTK, MMP-9, NFATc1, c-Fos), protein
expression of makers (NFATc1, c-Fos) were investigated.
Moreover, the expressions of MAPK, NF-κB, I-κB by RANKL were
studied using western blot analysis. For in vivo study, 40 female
Sprague-Dawley (SD) rats were divided into five groups: Sham
(Sham-operated group) and OVX (ovariectomized groups) were
medicated with distilled water, and the remaining ovariectomized
groups were orally administrated with E2 (17β-estradiol, 100 μ
g/kg), wFF-L (31 mg/kg), wFF-H (310 mg/kg) for 8 weeks. And
then body weight, uterus weight, bone density of femur, femurs
weight, tibia weight were measured respectively.
Results : wFF suppressed TRAP positive cell formation and
TRAP activity, Pit area formation in RANKL-induced RAW 264.7
cells. And wFF decreased the expression of
osteoclastogenesis-related markers (TRAP, CAⅡ, CTK. MMP-9)
and also reduced the protein expression of maker (NFATc1,
c-Fos). These suggest that wFF inhibits osteoclast differentiation
and bone resorption. In the OVX rat model, wFF inhibited
increasing of body weight, decreasing of BMD, decreasing of
femurs weight, decreasing of tibia ash and decreasing oftrabecular area.
Conclusions : From the results of the above, Forsythiae
Fructus should be effective for osteoporosis prevention and
treatment.
characterized by low bone mass and increased bone fragility. It
has become a major problem of senior citizens. Recently, many
studies of the bone remodelling process suggest that factors
involved in inflammation are linked with osteoporosis. The
purpose of this study is to experiment the effect of water extract
of Forsythiae Fructus (wFF) on osteoclast differentiation; and the
other purpose is to examine the effect of wFF on osteoporosis in
ovariectomized rat.
Materials and methods : To test the effect of wFF on
osteoclast differentiation and suppression of gene expression
related to bone resorption RAW 264.7 cells were used. Various
level of concentration of wFF (50, 100, 200 μg/mL) were added to
RAW 264.7 cells co-cultured with RANKL. The number of TRAP
positive cell, TRAP activity, Pit area, mRNA expression of makers
(RANK, TRAP, CAⅡ, CTK, MMP-9, NFATc1, c-Fos), protein
expression of makers (NFATc1, c-Fos) were investigated.
Moreover, the expressions of MAPK, NF-κB, I-κB by RANKL were
studied using western blot analysis. For in vivo study, 40 female
Sprague-Dawley (SD) rats were divided into five groups: Sham
(Sham-operated group) and OVX (ovariectomized groups) were
medicated with distilled water, and the remaining ovariectomized
groups were orally administrated with E2 (17β-estradiol, 100 μ
g/kg), wFF-L (31 mg/kg), wFF-H (310 mg/kg) for 8 weeks. And
then body weight, uterus weight, bone density of femur, femurs
weight, tibia weight were measured respectively.
Results : wFF suppressed TRAP positive cell formation and
TRAP activity, Pit area formation in RANKL-induced RAW 264.7
cells. And wFF decreased the expression of
osteoclastogenesis-related markers (TRAP, CAⅡ, CTK. MMP-9)
and also reduced the protein expression of maker (NFATc1,
c-Fos). These suggest that wFF inhibits osteoclast differentiation
and bone resorption. In the OVX rat model, wFF inhibited
increasing of body weight, decreasing of BMD, decreasing of
femurs weight, decreasing of tibia ash and decreasing oftrabecular area.
Conclusions : From the results of the above, Forsythiae
Fructus should be effective for osteoporosis prevention and
treatment.
목차
I. 서 론 ········································································································· 1II. 재료 및 방법 ······························································································ 41. 재료 ·········································································································· 41) 시약 ······································································································ 42) 시료의 준비 ························································································ 42. 세포 실험 방법 ······················································································ 51) RAW 264.7 세포 배양 ··································································· 52) 세포 독성 측정 ·················································································· 53) TRAP 염색 ························································································· 54) TRAP 활성도 측정 ············································································ 65) pit 형성 억제 측정 ········································································· 66) 역전사 중합 반응 분석 ····································································· 77) Western blot 분석 ··········································································· 83. 동물 실험 방법 ······················································································ 91) 실험동물 사육 및 관리 ····································································· 92) 난소절제술를 이용한 폐경기 골다공증 모델 제작 및 장기무게변화측정 ································································································ 93) 대퇴골 무게 측정 ············································································ 104) 폐경기 골다공증 흰쥐 대퇴골의 조직학적 검사 ························ 104. 통계분석 ································································································ 11III. 실험결과 ·································································································· 121. 連翹가 RAW 264.7 세포 생존율에 미치는 영향 ·························· 122. 連翹가 파골세포 형성 및 활성 억제에 미치는 효능 ···················· 143. 連翹의 pit 형성억제에 미치는 영향 ················································ 164. 連翹의 파골세포 기능에 필요한 유전자 발현 억제에 미치는 효능 ············································································································· 185. 連翹의 NFATc1, c-Fos 유전자 및 단백질 발현 억제에 미치는효능 ········································································································· 206. 連翹의 MAPK 인산화에 미치는 효능 ············································· 227. 連翹의 NF-κB, I-κB 발현에 미치는 효능 ····································· 248. 連翹의 동물 몸무게 및 장기무게에 미치는 효과 ························· 269. 連翹의 대퇴골 골소주 면적에 미치는 효과 ··································· 29IV. 고 찰 ······································································································ 31V. 결 론 ······································································································· 36참고문헌 ········································································································· 37ABSTRACT ···································································································· 43
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