초록 ▼

Ⅲ. 연구개발의 내용 및 범위 1. 인삼으로부터 사포닌 및 비사포닌 분획 분리 인삼의 면역기능 개선, 지질과산화 개선 및 에너지대사 개선 효능 등을 평가하기 위해 실험에 사용된 백삼과 홍삼은 농협에서 판매하는 것을 구입하여 사용하였다. 백삼과 홍삼을 각각 100g씩 70% 에탄올로 3회 환류 추출하여 감압∙농축한 다음, 증류수를 첨가하여 10% 추출물을 제조한 후, 부탄올 분획을 3회 실시하여 상층액을 농축∙건조하여 사포닌 분획을 얻었다. 사포닌분획은 백삼으로부터 8.8g, 홍삼으로부터 10.4g이 분리되었다. 2. 인삼 추출물의

Ⅲ. 연구개발의 내용 및 범위 1. 인삼으로부터 사포닌 및 비사포닌 분획 분리 인삼의 면역기능 개선, 지질과산화 개선 및 에너지대사 개선 효능 등을 평가하기 위해 실험에 사용된 백삼과 홍삼은 농협에서 판매하는 것을 구입하여 사용하였다. 백삼과 홍삼을 각각 100g씩 70% 에탄올로 3회 환류 추출하여 감압∙농축한 다음, 증류수를 첨가하여 10% 추출물을 제조한 후, 부탄올 분획을 3회 실시하여 상층액을 농축∙건조하여 사포닌 분획을 얻었다. 사포닌분획은 백삼으로부터 8.8g, 홍삼으로부터 10.4g이 분리되었다. 2. 인삼 추출물의 지방산 산화 억제 효과 연구 DCF-DA assay로 활성산소 제거능을 측정하였다. 지질 과산화 분석은 thiobarbituric acid (TBA) 반응법을 이용하였다. 인삼추출물에 의해 3T3-L1 세포가 분화하여 생성된 중성지방의 함량 및 지방분해능을 검사하였다. 항산화 효소의 발현은 인삼 추출물을 처리하여 total RNA을 분리한 후, cDNA을 합성하여 RT-PCR을 실행함으로써 확인하였다. 항효소의 활성 평가는 과산화수소 처리 전·후에 인삼시료를 처리함으로써, 세포내 항산화 효소의 활성 정도를 평가하였다. 3. 인삼 추출물의 항염증 효과 연구 RAW264.7 대식세포는 FBS (10%)과 penicillin (100 U/ml)와 streptomycin sulfate(100 μg/ml)가 함유된 DMEM배지에서 습한 조건의 37℃, 5% CO₂ Incubator에서 배양하였다. 본 실험에서 RAW264.7 대식세포에 대한 세포독성 및 실험 시 처리 농도를 결정하고, 인삼 추출물의 세포독성을 측정하기 위해 MTT assay를 사용하였다. 인삼추출물의 항염증 효과를 탐색하기 위해 NO (nitric oxide)생성량 측정하였다. 또한, 생쥐에서 분리한 비장세포 배양 상층액에서 IL-1beta, MCP-1 specific IgE를 ELISA kit을 사용하여 생산량을 측정하였다. 항염증 효능 기전을 구명하기 위해 염증세포에서 분리한 total RNA을 이용한 RT-PCR법과 단백질 분획물을 이용한 western blot법을 사용하였다. 4. 인삼 추출물의 에너지 대사 개선 효과 연구 에너지대사 개선 효과 연구를 위해 ATCC에서 구입한 L6 세포를 이용하여 실험하였다. DCF-DA법을 이용해 reactive oxygen의 생성량을 측정하였고, glucose uptake 측정을 위해 세포를 24 well culture dish에 seeding하여 완전히 분화시킨 후 시료를 세포배양액에 가하고 [³H]-2-deoxyglucose을 처리하여 radioactivity를 측정하였다. 에너지 대사 관련 단백질 발현 분석은 lysis buffer을 이용하여 단백질 분획을 추출한 후, western blot법으로 실행하였다. 5. 고려인삼의 인체적용 연구 고려인삼(백삼)의 인체적용 시험을 수행하기 전에 원료의 성분분석 및 안전성 검사를 우선 수행하였다. 고려인삼의 진세노사이드 분석은 UV검출기가 장착된 HPLC을 이용하였다. 잔류농약 분석은 인삼 시료를 전처리하여 Agilent사의 6890 GC-ECD를 사용하였다. 인삼시료에 함유된 병원성 미생물 (E.coli, S. aureus, 살모넬라)을 검사하였고, 곰팡이와 효모의 생균수를 측정하였다. Aflatoxin분석은 AOAC-CB법에 준하여 시행하였다. 인삼시료의 중금속 검사을 위해 습식 분해법인 킬달 분해법을 이용하여 전처리 하였고, Cr, Cu, Pb, Ni, Zn, Cd, Hg 등을 해당 분석법에 따라 함량을 검사하였다.

Abstract ▼

Obesity is an underlying factor associated with a variety of life-threatening diseases including diabetes mellitus, heart failure, cardiovascular diseases, and liver malfunction. Alarmingly, there is a high incidence of obesity in children and adolescents, indicating that obesity will be a significa

Obesity is an underlying factor associated with a variety of life-threatening diseases including diabetes mellitus, heart failure, cardiovascular diseases, and liver malfunction. Alarmingly, there is a high incidence of obesity in children and adolescents, indicating that obesity will be a significant health problem for the foreseeable future. The cellular basis of obesity has a direct causal link with high levels of triglycerides (TAGs), the primary fat in our bodies. Panax ginseng has been taken orally to improve physical strength by people in East Asia. Many scientific studies have shown that ginseng helps prevent diabetes mellitus, atherosclerosis, hyperlipidemia, hypercholesteremia, immune dysfunction among other disorders. Beneficial effect of Korean red ginseng on triglyceride synthesis and lipid differentiation is not still yet completely elucidated. Also, No attempts have been made to elucidate the comparative effects of white ginseng and red ginseng on triglyceride synthesis and lipid differentiation. Therefore, this study was carried out to compare the beneficial role of saponin fractions isolated from white ginseng (WG) and red ginseng (RG) on lipid accumulation in 3T3-L1 preadipocytes. This study was carried out to determine and compare inhibitory effects of white ginseng (WG) and red ginseng (RG) on 3T3-L1 adipocyte differentiation. Ginseng was extracted with 70% methanol and partitioned with butanol to obtain saponin fractions, which have been known as bioactive constituents of ginseng. Triglyceride assay, Oil-red O staining, lipolysis assay were performed to elucidate beneficial effect of ginseng on in 3T3-L1 preadipocyte differentiation. As a result, all 70% MeOH extracts and saponin fraction of RG decreased triglyceride content accounting for 36.7±6.8 μM (p < 0.01). Consistent with this result, lipid droplets in 3T3-L1 adipocytes were definitely decreased by treatment with saponin fraction of RG at a concentration of 100 μg/mL. Non-saponin fractions of both WG and RG not inhibited triglyceride synthesis and lipid differentiation. In contrast, triglyceride contents of the saponin fractions of WG and RG accounted for 99.4 ± 14.2 μM (p < 0.05) and 36.7 ± 6.8 μM (p < 0.01), respectively. Consistent with this result, lipid droplets in 3T3-L1 adipocytes were definitely decreased by treatment with saponin fractions of ginseng at a concentration of 100 μg/mL. Interestingly, saponin fraction of RG was approximately 2.5-fold more potent than that of WG. These results indicate that the saponin constituents of Panax ginseng considerably reduce triglyceride synthesis and adipocyte differentiation in 3T3-L1 cells. In especial, Korean red ginseng may certainly contribute to prevent introducing various vascular diseases during obesity development. Furthermore, red ginseng may certainly contribute more effectively than white ginseng to prevent introducing various vascular diseases during obesity development. However, further studies are needed to assure the potential effect of Korean red ginseng on adipogenesis.
The generation of reactive oxygen species (ROS) or free radicals can lead to cell and tissue damage paralleled by changes in the function of the genetic molecules, resulting in impaired oxygen delivery to the skeletal muscle and increased fatigue during physical activity. Enzymatic antioxidants like superoxide dismutase (SOD), which eradicates superoxide in presence of copper and zinc; glutathione peroxidase (GPx), which converts hydrogen peroxide into water and converts various hydroperoxides into less harmful hydroxides in presence of selenium; catalase (CAT), which can also break down hydrogen peroxide as well as non-enzymatic antioxidants such as vitamin C, vitamin E and reduced glutathione (GSH) scavenge ROS and free radicals or prevents their formation. Oxidative damage by oxygen free radicals such as superoxide anions are known to be one of the factors involved in the mechanisms of diseases including cardiovascular dysfunctions, atherosclerosis, inflammation, carcinogenesis, reperfusion injury. Oxygen free radicals are created through aerobic metabolism and are mostly removed by endogenous antioxidants such as SOD, GPx, GR and catalase. Antioxidant enzymes and antioxidant molecules are believed to play an important role in the prevention of oxidative stress-related diseases such as cancer, cardiovascular disease, Alzheimer’s disease and muscular degeneration. Therefore, we investigated the effects of Korean white and red ginsengs on oxidative stress in HepG2 hepatoma and L6 skeletal muscle cells. Malondialdehyde (MDA) content and 2',7'-dichlorofluorescein diacetate (DCF-DA) assay was measured for evaluating intracellular reactive oxygen species (ROS) generation. Also, mRNA expressions and activities of intracellular antioxidant enzymes were evaluated after treatment of saponin or non-saponin fractions of ginseng in H₂O₂-treated HepG2 and L6 cells. According to DCF-DA assay, ROS generation by H₂O₂ in HepG2 cells was significantly reduced more than 40% by treatment with ginseng extracts. ROS generation was significantly increased more than 2.8-fold in H₂O₂-treated L6 myotubes. Treatment of ginseng definitely attenuated MDA production by increasing lipid peroxidation in H₂O₂-treated HepG2 cells. In addition, Panax ginseng increased effectively intracellular antioxidant enzyme activities including catalase, glutathione peroxidase (GPx) and superoxide dismutase (SOD) in HepG2 cells. Interestingly, antioxidantive effect of ginseng showd at sponin fraction as well as non-sponin fraction of white or red ginseng. According to DCF-DA assay, ROS generation was significantly increased more than 2.8-fold in H₂O₂-treated L6 myotubes. In contrast, saponin fractions fo ginsng extracts effectively reduced ROS production to almost control level. Treatment of ginseng definitely attenuated MDA production by increasing lipid peroxidation in H₂O₂-treated L6 myotubes. In addition, Panax ginseng increased effectively intracellular antioxidant enzyme activities including catalase, GPx and SOD in L6 myotubes. Interestingly, antioxidative effect of red ginseng showed more potent than those of white ginseng. These results indicate that administration of Panax ginseng may certainly contribute to prevent from damaging skeletal muscle cells and liver function by oxidative stress.

목차 Contents

• 표지 ... 1
• 제출문 ... 2
• 요약문 ... 3
• SUMMARY ... 8
• 목 차 ... 12
• 제1장 서 론 ... 13
• 제1절 연구개발의 목적 및 필요성 ... 13
• 제2절 연구범위 ... 16
• 제2장 국내외 기술개발 현황 ... 17
• 제1절 국내외 연구 현황 ... 17
• 제2절 국내외 연구현황 비교 및 필요 연구 분야 ... 18
• 제3장 연구개발수행 내용 및 결과 ... 20
• 제1절 연구방법 및 내용 ... 20
• 제2절 연구내용 및 주요결과 ... 28
• 제4장 연구개발목표 달성도 및 대외기여도 ... 47
• 제1절 목표대비 대외 달성도 ... 47
• 제2절 정량적 성과 ... 48
• 제5장 연구개발결과의 활용계획 ... 50
• 제6장 연구개발과정에서 수집한 해외과학기술정보 ... 51
• 제7장 기타 중요 변동사항 ... 52
• 제8장 국가과학기술종합정보시스템에 등록한 연구장비 현황 ... 53
• 제9장 참고문헌 ... 54