표제지
목차
ABSTRACT 7
I. 서론 13
II. 재료 및 방법 17
1. Plant extract 17
2. Cell culture 18
3. Thiobarbituric acid reactive substance (TBARS) analysis 19
4. Alamar blue assay 20
5. Oil Red-O staining 21
6. Measurement of GPDH activity 22
7. RT-PCR for the mRNA expression of C/EBPα, and PPARγ 23
8. Statistical analysis 23
III. 결과 및 고찰 24
1. 천궁 추출물의 항산화 활성 24
가. Thiobarbituric acid reactive substance (TBARS) analysis 24
2. 3T3-L1 및 C2C12 세포를 이용한 지방분화조절 및 세포독성 및 지방분화 활성 27
가. Alamar blue assay를 이용한 세포독성 조사 27
나. 천궁 메탄올추출물 처리에 따른 GPDH 활성변화 29
다. 천궁 메탄올추출물 처리에 따른 3T3-L1 지방전구세포의 분화 활성: Oil red O 30
라. 천궁 메탄올추출물 처리에 따른 PPARγ and C/EBPα 유전자의 mRNA 발현조사 35
3. 비타민A, 비타민C 및 천궁 메탄올추출물의 단일 또는 혼합처리에 의한 지방세포 분화활성 변화 37
가. 비타민A, 비타민C 및 천궁 메탄올추출물의 단일 또는 혼합처리에 의한 3T3-L1 지방전구세포의 분화활성 변화: Oil Red O 37
나. 비타민A, 비타민C 및 천궁 메탄올추출물의 단일 또는 혼합처리에 의한 3T3-L1 지방전구세포의 분화활성 변화: GPDH Assay 41
다. 비타민A, 비타민C 및 천궁 메탄올추출물의 단일 또는 혼합처리에 의한 3T3-L1 지방전구세포의 PPARγ 및 C/EBPα 유전자 mRNA 발현조사 44
4. 천궁 메탄올추출물의 지방세포분화 조절 작용기전 48
가. 지방세포의 분화유도인자의 처리조건에 따른 GPDH 활성 48
IV. 적요 52
V. 참고문헌 54
Fig 1. Effect of different concentration of quercetin on oxidant -induced TBARS production in brain homogenates measured using micro-plate reader set at535 nm wave length. Br=brain homogenate only; Ox=oxidant (10 μM Aluminum Ammonium... 25
Fig 2. Relationship between quercetin concentration and mean TBARS OD at 535 nm (A), and the inhibition rate (%) of oxidant-induced TBARS production of brain homogenate by different concentrations of quercetin (B). Means with different... 26
Fig 3. Antioxidant activity of Cnidium officinale methanol extract against lipid peroxidation expressed as quercetin equivalents. 26
Fig 4. Effects of different concentration of Cnidium officinale methanol extracts on viability of mouse 3T3-L1 preadipocyte cells (a) and C2C12 fibroblast cells (b) measured by Alamar blue assay method after 24h and 48h post-treatment.... 28
Fig 5. Effects of different concentration of Cnidium officinale methanol extracts on GPDH activities of mouse 3T3-L1 preadipocyte cells 29
Fig 6. Effects of different concentration of Cnidium officinale methanol extracts on the differentiation of mouse 3T3-L1 preadipocyte cells; (a) Photographs of differentiated 3T3-L1 cells taken by microscope before Oil red O... 34
Fig 7. Effects of different concentration of Cnidium officinale methanol extracts on mRNA expressions of PPARγ and C/EBPα genes of mouse 3T3-L1 preadipocyte cells measured by RT-PCR 36
Fig 8. Photographs of differentiated 3T3-L1 adipocytes under the treatment of vitamin A or vitamin C with/out Cnidium officinale methanol extracts measured by Oil Red O method 39
Fig 9. Optical density of Oil Red O staining from differentiated 3T3-L1 adipocyte cells under the treatment of vitamin A or vitamin C with/out Cnidium officinale methanol extracts measured by Oil Red O method. 40
Fig 10. GPDH activity of lysates from differentiated 3T3-L1 preadipocytes under the treatment of vitamin A or vitamin C with/out Cnidium officinale methanol extracts 43
Fig 11. Relative mRNA expressions of PPARγ (upper) and C/EBPα (lower) in 3T3-L1 adipocytes treated with single or combinations of vitamin A, vitamin C or CO methanol extract during differentiation. 47
Fig 12. Effects of insulin (A), dexamethasone (B) or IBMX (C) deprivation on 3T3-L1 differentiation treated with or without 100 μg/mL CO methanol extract . 51