보고서 정보
주관연구기관 |
전북대학교 Chonbuk National University |
보고서유형 | 최종보고서 |
발행국가 | 대한민국 |
언어 |
한국어
|
발행년월 | 2016-02 |
과제시작연도 |
2015 |
주관부처 |
농촌진흥청 Rural Development Administration(RDA) |
등록번호 |
TRKO201600003085 |
과제고유번호 |
1395041107 |
사업명 |
농축산물부가가치향상 |
DB 구축일자 |
2016-06-25
|
DOI |
https://doi.org/10.23000/TRKO201600003085 |
초록
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Ⅳ. 연구개발결과
○ 1세부과제명 : 세포주 및 실험동물모델을 이용한 팥의 비만/당뇨 개선효과와 작용기전규명
- 20여 종의 팥 에탄올 추출물에서 항산화 활성, 혈전용해효소 활성 스크리닝 후 선별된 팥 품종으로 지방세포, 췌장세포에서 팥 추출물의 비만/당뇨 효능 검증
- 고지방식이로 비만/당뇨 유발 실험동물을 이용, 팥 추출물 섭취시 혈중 및 간중 지질수준개선, 체중 증가 억제, 지방조직 축적 감소, 지방간 및 지질대사 조절 효소 유전자 발현 개선 등 비만개선 효과 확인
- 사육 중 주별 공복혈당 저하, 혈중
Ⅳ. 연구개발결과
○ 1세부과제명 : 세포주 및 실험동물모델을 이용한 팥의 비만/당뇨 개선효과와 작용기전규명
- 20여 종의 팥 에탄올 추출물에서 항산화 활성, 혈전용해효소 활성 스크리닝 후 선별된 팥 품종으로 지방세포, 췌장세포에서 팥 추출물의 비만/당뇨 효능 검증
- 고지방식이로 비만/당뇨 유발 실험동물을 이용, 팥 추출물 섭취시 혈중 및 간중 지질수준개선, 체중 증가 억제, 지방조직 축적 감소, 지방간 및 지질대사 조절 효소 유전자 발현 개선 등 비만개선 효과 확인
- 사육 중 주별 공복혈당 저하, 혈중 인슐린 및 인슐린 저항성 지표 정상화 등내당능 장애 개선 효과 확인
- 실험동물에서 식이와 함께 팥 추출물 공급할 경우 식이섭취량 감소에 영향 확인하였으며, 뇌 시상하부 식욕 조절 유전자 발현이 식욕감소, 포만 유도로 유의적 차이 확인
- 팥 추출물을 음료 제조에 용이하도록 침전물 형성 방지 공정 확립, 배합비 개발, 제조공정 확립, 발효음료 안정성 확인 후 대량생산 공정과 생산단가 절감연구 수행하여 팥 싹 분말 3종, 유산균발효음료, 환 등 제품 개발
○ 2세부과제명 : 실험동물을 이용한 팥의 혈행개선 효능 검증 및 작용기전 연구
- 실험동물 식이와 팥 추출물을 혼합하여 공급한 후 혈액 응고시간, 혈전 용해능(Fibrinogen, FDP, D-dimer), 혈소판 응집능, 혈소판 수를 측정하고 tail bleeding test를 통해 혈행개선 효능 검증
○ 1협동과제명 : 싹나물과 발효음료용으로 가공적성이 우수한 팥 품종 선발
- 나물용으로 가공적성이 우수하고 다수성인 팥 우량계통 선발
- 발아시간에 따른 팥 싹나물의 생육길이 및 수율, 일반성분 및 무기성분, 총 폴리페놀, 플라보노이드 및 탄닌 함량 분석
- 발아시간에 따른 팥 싹나물 에탄올 추출물의 항산화활성 분석
- 발효음료 제조에 적합한 팥 품종 선별을 위해 당도, 탁도, 항산화 활성 확인
- 팥 싹나물 분말 가공제품의 일반 및 무기성분, 기능성 성분, 항산화 활성 측정
- 팥 유산균 발효음료 가공제품 품질 특성 및 항산화 활성 확인
○ 2협동과제명 : 팥 추출물로부터 활성 물질 분리 및 효능검색
- 팥 분획물의 항산화 활성 확인 후 분획물에서 화합물을 분리하여 항산화 효능 검증을 하였으며, genistein, kaempferol, quercetin, rutin 등 6종의 화합물을 분리하였음
- 팥에서 분리한 genistein이 Caenorhabditis elegans(선충)에서 수명연장, 스트레스 저항성 증가, 항산화 효소활성 증가, 체내 SOD-3, HSP-16.2 발현 효과 확인
- 팥에서 분리한 vitexin이 Caenorhabditis elegans(선충)에서 수명연장, ROS 축적억제, 항산화 효소 활성 증대, 열 및 산화적 스트레스 저항성 효과 확인하였으며, 선충의 활동능 증가를 보여줌
- 팥나물 추출물의 항산화효과 확인
Abstract
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1. Investigation of anti-obesity/diabetes effect of adzuki bean (Vigna angularis) and its underlying mechanism in vitro and in vivo
Effects of adzuki bean (Vigna angularis) extract on proliferation and differentiation of 3T3-L1 preadipocytes into mature adipocytes; The aim of this work was to inv
1. Investigation of anti-obesity/diabetes effect of adzuki bean (Vigna angularis) and its underlying mechanism in vitro and in vivo
Effects of adzuki bean (Vigna angularis) extract on proliferation and differentiation of 3T3-L1 preadipocytes into mature adipocytes; The aim of this work was to investigate the effects of black adzuki bean (BAB) extract on adipocytes, and to elucidate the cellular mechanisms. In order to examine the proliferation of preadipocytes and differentiating adipocytes, cell viability and DNA content were measured over a period of time. Lipid accumulation during cell differentiation and the molecular mechanisms underlying the effects of BAB on the transcriptional factors involved, with their anti-adipogenic effects, were also identified. We observed that BAB exhibits anti-adipogenic effects through the inhibition of proliferation, thereby lowering mRNA expression of C/EBPβ and suppressing adipogenesis during the early stage of differentiation. This, in turn, resulted in a reduction of TG accumulation in a dose- and time-dependent manner. Treating the cells with BAB not only suppressed the adipogenesis-associated key transcription factors PPARγ and C/EBPα but also significantly decreased the mRNA expression of GLUT4, FABP4, LPL and adiponectin.The expression of lipolytic genes like ATGL and HSL were higher in the treatment group than in the control. Overall, the black adzuki bean extract demonstrated an anti-adipogenic property, which makes it a potential dietary supplement for attenuation of obesity.
Adzuki bean (Vigna angularis) inhibits obesity through the modulation of lipid metabolism in diet induced obese mice; The primary objective of this study was to investigate the obesity inhibitory effects of black adzuki bean (BAB) extract on high-fat diet induced obese mice. For 12 weeks, 4-week-old C57BL/6J mice were fed a control diet containing 10kcal% from fat, a high-fat diet containing 60kcal% from fat (HD), a high-fat diet mixed with 1% freeze-dried powder BAB extract (BAB), or a high-fat diet mixed with 0.08% kaempferol (Kfr). Compared to the HD group, the BAB group showed significantly reduced body weight, reduced white adipose tissue weight, and an adipocyte phenotype. BAB decreased signs of liver pathology, liver weight, and the formation of hepatic lipid droplets. Lipogenesis-associated genes were significantly suppressed in epididymal fat tissue, while elevated levels of lipolytic and β-oxidation genes were observed in the BAB group relative to the HD group. Overall, BAB treatment is demonstrated as a potential dietary supplement capable of attenuating obesity.
Adzuki bean extract (Vigna angularis) protects pancreatic cells and ameliorate glucose tolerance in C57BL/6J mice fed a high fat diet; In the present study, we investigated the effect of black adzuki bean (BAB) extract on pancreatic cells and determined their mechanism of action in impaired glucose tolerance in an animal model of type 2 diabetes. In addition, we performed, functional gene annotation analysis to identify genes related to the regulation of glucose metabolism and insulin response. Treatment of pancreatic β-cells with BAB (0.2 mg/mL) led to tolerance of the high glucose induced glucotoxicity, resulting in a similar viability as cells maintained in normal glucose media.In addition, dietary supplementation with BAB significantly (P < 0.05) improved hyperglycemia and HOMA-IR in high fat diet induced glucose intolerance obese C57BL/6J mice. Our results suggest that BAB extract ameliorates hyperglycemia, glucose intolerance, and lowers HOMA-IR by regulating insulin secretion and response, and maintain the integrity of pancreatic β-cells from hyperglycemic conditions.
Effects of adzuki bean (Vigna angularis) extract on body composition and hypothalamic neuropeptide expression in rats fed a high-fat diet; Adzuki beans have been evaluated as potential remedies for various health conditions, and recent studies have reported their effects on the regulation of lipid metabolism, but it remains to be determined whether they may be effective in overcoming obesity by regulating appetite and satiety. This study investigated the effect of black adzuki bean (BAB) extract on body composition and hypothalamic neuropeptide expression in Sprague Dawley rats(Rattus norvegicus) fed a high-fat diet. The body weights and epididymal fat weights were significantly reduced and the serum lipid profiles were improved in the group fed the diet containing BAB compared to the HD group. The expression of AGRP mRNA significantly decreased in the BAB groups, and treatment with BAB-2 resulted in a marked induction of the mRNA expression of POMC and CART, which are anorexigenic neuropeptides that suppress food intake. Furthermore, mRNA expression levels of ObRb, a gene related to leptin sensitivity in the hypothalamus, were significantly higher in the BAB groups than in the HD group. These results suggest that supplementation with BAB has a significant effect on body weight via regulation of hypothalamic neuropeptides.
2. Adzuki Beans (Vignaangularis) supplementation improves blood flow in high fat diet fed Rats
This study investigated the effects of black adzuki bean (BAB) on the in vitro fibrinolytic activity and in vivo blood flow improving effect in SD rats. We evaluated the fibrinolytic activity of 20 different kinds of adzuki bean ethanol extract and BAB exhibited effective fibrinolytic activity compared to the rest of the adzuki beans. For in vivo study, four weeks old SD rats were fed for 8 weeks with normal diets containing 10% kcal from fat (ND) or high-fat diets containing 60% kcal from fat (HD) or a high fat diet+1% freeze-dried ethanolic extract of black adzuki bean (BAB-1). BAB supplementation resulted in normalization in serum triglyceride (TG), total cholesterol(TC), fibrin degradation product (FDP), fibrinogen, adenosine diphosphate (ADP) and platelet (PLT). Overall, these results suggest that the black adzuki bean extract demonstrated a blood flow improving property, but we need further study to makes a potent dietary supplement for anti-platelets.
3. The purpose of this study was to evaluate the yield, antioxidant content, and antioxidant activity of adzuki beans according to germination time. Cultivated varieties were Vigna angularis var. Nipponensis cv. Chungju-pat(CJP), and Yeonduchae (YDC), and Vigna radiata (L.) R. Wilczek cv. Dahyeon (DH). The moisture, crude protein, calcium, and magnesium contents of YDC significantly changed with increasing germination time, whereas potassium, natrium, and aluminium contents did not significantly change. Sprout yield, total polyphenol, flavonoid, and tannin contents of ethanolic extracts from adzuki and mung beans significantly increased with increasing germination time.
Total polyphenol contents of ungerminated CJP, YDC, and DH were 1.96, 2.68, and 2.02mg/g, and those of CJP and YDC germinated for 144 h were 3.33 and 3.47 mg/g, respectively. Total flavonoid content of adzuki beans substantially decreased with increasing germination time. Total tannin content substantially increased with increasing germinationtime, and YDC showed higher contents (0.85 mg/g) sample germinated for 120 h. DPPH radical scavenging activities of CJP and YDC substantially decreased with increasing germination time, whereas that of DH increased. ABTS radical scavenging activities of ungerminated CJP, YDC, and DH were 4.57, 6.51, and 2.82 mg/g,respectively, and increased after germination for 72∼120 h.
We evaluated the quality characteristics of adzuki bean fermented beverages on the breeding lines. The total polyphenols of the adzuki bean fermented beverages races after germination for 24 and 48 h were 149.16∼435.68 and 167.80∼413.40, respectively.The flavonoid contents of the adzuki bean fermented beverages races after germination for 24 and 48 h were 19.04∼33.61 and 21.63∼41.21, respectively. The ABTS radical scavenging activity of the adzuki bean fermented beverages races after germination for 24 and 48 h were 4.74∼8.50 and 3.87∼9.96 respectively.
4. Effects of genistin and vitexin from Vigna angularis on Lifespan-extending in Caenorhabditis elegans
Several theories emphasize that aging is closely related to oxidative stress and disease. The formation of excess ROS can lead to DNA damage and the acceleration of aging. Vigna angularis is one of the important medicinal plants in Korea. Previous phytochemical studies of Vigna angularis (Ohwi) Ohwi & Ohashi (Leguminosae) have shown the presence of saponins and flavonoids. From the seed of V. angularis,genistein-7-O-β-D-glucopyranoside (genistin) and vitexin were isolated. Lifespan-extending effect of genistin and vitexin were elucidated using Caenorhabditis elegans model system. Genistin showed potent lifespan extension of worms under normal culture condition. This compound also exhibited the protective effects against thermal and oxidative stress conditions. In the case of heat stress, genistin-treated worms exhibited enhanced survival rate, compared to control worms. In addition, genistin-fed worms lived longer than control worms under oxidative stress induced by paraquat. To verify the possible mechanism of genistin-mediated increased lifespan and stress resistance of worms, we investigated whether genistin might alter superoxide dismutase (SOD), catalase activities and intracellular ROS levels. Our results showed that genistin was able to elevate SOD and catalase activities of worms and reduce intracellular ROS accumulation in a dose-dependent manner. Vitexin showed potent lifespan extensive activity and it elevated the survival rates of nematodes against the stressful environ\-ments including heat and oxidative conditions. In addition, our results showed that vitexin was able to elevate antioxidant enzyme activities of worms and reduce intracellular ROS accumulation in a dose-dependent manner. These studies demonstrated that the increased stress tolerance of vitexin-mediated nematode could be attributed to increased expressions of stress resistance proteins such as superoxide dismutase (SOD-3) and heat shock protein (HSP-16.2). In this work, we also studied whether vitexin-mediated longevity activity was associated with aging-related factors such as progeny, food intake, growth and movement. The data revealed that these factors were not affected by vitexin treatment except movement. Vitexin treatment improved the body movement of aged nematode, suggesting vitexin affects healthspan as well as lifespan of nematode. These results suggest that vitexin might be a probable candidate which could extend the human lifespan.
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