효소를 이용한 감태분획물의 항비만 효과

다국어 초록 (Multilingual Abstract)

Obesity is a worldwide problem that contributes to serious diseases including diabetes, hypertension and atherosclerosis. Recently, much research is conducted about functional natural materials and their anti-obesity activity. This study investigated the effects of the enzyme-treated Ecklonia cava extracts (EEc) on 3T3-L1 preadipocytes and mice fed high fat diet.
In vitro, EEc in order to observe effect of inhibiting adipogenesis, EEc were treated with different concentrations (12.5 μg/ml, 50 μg/ml, 200 μg/ml) on induced differentiation of 3T3-L1 cells. The results of MTS assay were not observed that cell viability reduced on any concentrations of EEc treatment. The levels of glucose in differentiated 3T3-L1 cells were lowest in the group treated with a concentration of 200 μg/ml EEc. Triglyceride accumulations and lipid droplets were decreased in a dose-dependent manner. In addition, EEc reduced the C/EBPs transcriptional activity significantly in 3T3-L1 cells.
In vivo, to test anti-obese effect of diet containing EEc, C57BL/6NTac mice were divided into six groups : Normal diet group (ND), High fat diet group (HFD), High fat with Garcinia extract diet group (GHD), High fat with EEc diet group (EHD250, EHD500, EHD1000), and all groups were fed with each diet for 9 weeks. After the end of all feeding, factors relevant to lipid metabolism which body weight, lipid patterns, concentration of glucose and leptin in serum were determined, also GOT, GPT were measured, and protein expression in liver tissue were determined by western blot analysis. Body weight was significantly increased in HFD group compared to EHD groups, and the weights of liver, perirenal fat and epididymal fat corresponded to the increase in body weight. The levels of triglyceride, total cholesterol, low density lipoprotein-cholesterol in blood serum were lower in the EHD1000 group compared to the HFD group. Glucose and leptin concentrations were lowest in the EHD1000 group. The levels of GOT and GPT in the EHD1000 group were lower than in the HFD group and C/EBPs expressions were decreased in a dose-dependent manner. These results demonstrate that EEc significantly inhibit adipogenesis and can be used for the regulation of obesity.

목차 (Table of Contents)

• Ⅰ. 서론 1
• Ⅱ. 재료 및 방법 4
• 1. 실험 재료
• 1) 시약 및 재료 4
• 2. 실험방법 5
• 1) 효소를 이용한 감태분획물 (EEc)의 제조 5
• 2) 세포실험 5
• ① 3T3-L1 preadipocyte 배양 및 MTS assay 5
• ② 세포 내 glucose 및 triglyceride 함량 6
• ③ Oil red O 염색 7
• ④ Western blot 분석 7
• 3) 동물실험 8
• ① 실험동물의 사육 및 식이조성 8
• ② 실험동물의 혈액 및 장기 회수 11
• ③ 체중변화, 식이섭취량 측정 및 식이효율 계산 11
• ④ 혈청 지질 함량 측정 11
• ⑤ 혈청 glucose 및 leptin 농도 측정 12
• ⑥ 혈청 중의 GOT, GPT 측정 12
• ⑦ 간 조직 내 triglyceride 함량 측정 12
• ⑧ Western blot analysis 13
• 4) 통계학적 처리 13
• Ⅲ. 결과 및 고찰 15
• 1. 세포실험 결과 15
• 1) 3T3-L1 preadipocyte의 세포생존율 15
• 2) 세포 내 glucose 함량의 변화 17
• 3) 세포 내 triglyceride 함량의 변화 19
• 4) Lipid droplet 생성에 미치는 영향 21
• 5) 세포 수준에서 adipogenesis 저해활성 23
• 3. 동물실험 결과 25
• 1) 체중변화, 식이섭취량 및 식이효율 25
• 2) 장기 및 주변지방량의 변화 28
• 3) 혈청 지질의 변화 30
• 4) 혈청 glucose 및 leptin 농도의 변화 32
• 5) 혈청 중의 GOT, GPT 활성 변화 36
• 6) 간 조직 내 triglyceride 함량의 변화 38
• 7) 간 조직에서 adipogenesis 저해활성 40
• Ⅳ. 결론 및 요약 42
• Ⅴ. 참고 문헌 45