초록 ▼

○ 연구결과
① 돌나물 유전자원 보존 및 저장성 향상 - RAPD를 이용한 유연관계 분석결과, 2군으로 분류할 수 있었으며, Ⅰ군에는 완주 및 군산 지역종을 포함한 27개 지역종이 속하였고, Ⅱ군에는 줄기와 잎이 가늘고 꽃이 잘 피는 금산, 부천, 포천, 용인 등 4 지역종이 속하였으며, 형태적 특성과 잘 일치하였음.
- 기내 저온 활성보존은 0.2mg/L BA와 50mg/L sucrose, 1.2% agar를 기본으로 한 배지에 10-20mg/L ABA 또는 6-9% sucrose를 첨가하는 것이 양호하였음. 초저온 동

○ 연구결과
① 돌나물 유전자원 보존 및 저장성 향상 - RAPD를 이용한 유연관계 분석결과, 2군으로 분류할 수 있었으며, Ⅰ군에는 완주 및 군산 지역종을 포함한 27개 지역종이 속하였고, Ⅱ군에는 줄기와 잎이 가늘고 꽃이 잘 피는 금산, 부천, 포천, 용인 등 4 지역종이 속하였으며, 형태적 특성과 잘 일치하였음.
- 기내 저온 활성보존은 0.2mg/L BA와 50mg/L sucrose, 1.2% agar를 기본으로 한 배지에 10-20mg/L ABA 또는 6-9% sucrose를 첨가하는 것이 양호하였음. 초저온 동결보존은 0.4-0.6M sucrose 용액에서 1일간 전배양 후 냉동보존할 경우, 재생율은 11.1-16.7%였음.
- 돌나물을 스치로폼 접시에 랩포장 할 경우, 5℃에서는 수확 후 8일까지 상품성을 유지한 상태로 저장이 가능하였음.
② 돌나물의 건강 기능성 구명 및 고부가가치 상품의 개발
- Ethyl acetate 추출물로부터 간기능 개선 및 간암억제 화합물은 tetrahydroxyflavonol(quercetin)로 동정되었으며, 돌나물 추출물은 사염화탄소에 의한 급성 간 손상을 재생시키는데, silymarin(대조구)보다 효과적이었음.
- 돌나물 추출물은 ethionine 및 ethanol에 의한 지방간 유도를 유의적으로 억제하였고, 특히 hexane과 ethyl acetate 추출물에서 지방간 억제효과가 매우 우수하였으며, 간암 세포주 HepG2의 성장에 미치는 ethyl acetate 추출물의 IC50은 155.8 μg/mL로 세포 성장 억제 효과가 가장 우수하였음.
돌나물 즙과 을 백서에 - ethanol 경구투여한 후 2시간이 지났을 때 혈중 ethanol 농도는 대조구보다 약 2.3배 낮아 ethanol을 분해가 우수하였고, 돌나물의 methanol 및 butanol 추출물은 ADH 및 ALDH 효소활성을 유의적으로 증가시켰음.
- 백서에 ethanol 경구투여 전에 돌나물 추출물 투여구에서는 1시간 경과시 혈중 acetalde-hyde의 농도가 대조구의 약 50%로 시간이 경과하면서 빠르게 감소하였고, 인체에서도 혈중 ethanol 및 acetaldehyde의 농도를 유의적으로 감소시켰음.
- 이에 따라 돌나물 추출물을 주원료로 하여 숙취해소 음료를 제조하였음.
- 돌나물 ethyl acetate 추출물의 subfraction E4의 항산화활성이 가장 우수하였으며, 이 물질은 quercetin으로 동정되었고, 면역활성은 ethyl acetate 추출물 subfraction E5에서 가장 우수하였으며, 이 물질은 coumarin계의 esculetin인 것으로 확인되었음.
- 백서에 돌나물 추출물을 4주간 섭취시킨 간조직 및 혈액에서의 효소활성은 양성대조구인 tocopherol 또는 ascorbic acid와 유사하거나 우수한 항산화능을 나타냈고, 특히 methanol 및 ethyl acetate 추출물에서 polyphenol 함량이 높게 나타났음.
- 백서에 고지방식을 급여한 후 돌나물 추출물 처리는 항산화 관련 glutathione peroxidase, glutathione reductase의 발현증가와 면역기능 관련 COX2 단백질의 발현을 증가시켰음.
- 돌나물 즙(juice)에 당과 겔화제를 혼합, 성형하여 젤리를 제조하였으며, 돌나물 즙(juice)의 참가농도에 따라 칼슘, 철분, 칼륨, ascorbic acid 및 유기산 함량이 증가되었음.
- 돌나물 추출물의 지방세포의 성장을 억제하는 효과는 ethyl acetate subfraction E1의 효과가 가장 우수하였으며, 지방 세포 성장을 억제하는 물질은 caffeic acid로 동정되었음.
- 지방전구세포 MC3T3-L1이 지방세포로 분화되는 것을 50% 억제시키는 돌나물 ethyl acetate 추출물의 IC50은 66.98 μg/mL이었고, 유전자들의 발현양을 측정한 결과, PPAR-gamma는 ethyl acetate 추출물을 첨가시 거의 발현되지 않았음.
- 백서의 고콜레스테롤 식이에서 돌나물 분말 급여량이 많을수록 식이섭취량이 많았음에도 체중 증가량은 크게 감소하였고, total cholesterol, TG 및 LDL 농도 및 동맥경화지수 (AI)가 유의하게 감소하였으며, 간조직에 축적된 지방 및 혈청 ALP 농도도 대조구보다 낮았음.
- 돌나물 분말이 고지방식으로 유도된 비만, 고지혈증, 비알코올성지방간질환에 대한 효과가 우수하였으므로 돌나물 분말을 이용하여 돌나물 차를 개발하였음.
③ 개발된 제품의 안전성, 안정성 구명
- 돌나물의 무기이온은 군산, 금산, 완도 지역종의 칼륨, 칼슘, 철분 함량이 높았으며, ascorbic acid와 beta-carotene 함량은 군산 지역종에서 가장 높았음. 향기성분은 총 16종이 분리되었는데, pyrazine 및 이들 유도체가 주성분이었음.
- 돌나물을 이용한 숙취해소 음료, 항산화 및 면역활성 강화 젤리, 지질대사 및 혈행개선 차제품의 안전성 평가결과, 인체에 독성을 유발하지 않는 안전한 제품이었고, 음료는 실온에서 3개월, 차는 6개월, 젤리는 4℃에서 10일 동안 품질이 변하지 않아 안정성이 있었음.

Abstract ▼

One of the genus Sedum of Crassulaceae, S. sarmentosum Bunge is widely distributed in North-East Asia. S. sarmentosum is one of the favorite fresh-vegetables in Korea. The whole plant of S. sarmentosum has been traditionally used as a folk medicine for clinic of chronic viral hepatitis, skin burn, a

One of the genus Sedum of Crassulaceae, S. sarmentosum Bunge is widely distributed in North-East Asia. S. sarmentosum is one of the favorite fresh-vegetables in Korea. The whole plant of S. sarmentosum has been traditionally used as a folk medicine for clinic of chronic viral hepatitis, skin burn, and cancer in Korea. The plant has several alkaloids and terpens as major constituents, and its anti-hepatoxic property was also strong. Recently, S. sarmentosum was known as a source of herbal estrogen. In China, it is also used as a general gastric and renal regulator using various extracts or powders. However, S. sarmentosum has a low storage potential, and any healthy-food is not developed in Korea. Accordingly, there is an urgent need to work related to functional mechanisms and metabolism for producing health-benefit function food with high additive value in S. sarmentosum.
This work was conducted with the following objectives;
i) To study genetic relationship and in vitro conservation or cryopreservation for genetic resources and the storage temperature of shoot products for marketing of S. sarmentosum; ii) To develope the health-benefit function food based on the works related to functional mechanisms and metabolism for increasing the utilization and additive value of S. sarmentosum; iii) To investigate safety and stability of products for commercializing health- benefit function food using S. sarmentosum.
1. Anaysis of genetic relationship, in vitro conservation of genetic resources and development of shoot storage technique in S. sarmentosum
○ Thirty-one strains were classified into two groups by cluster analysis based on the morphological characteristics, and Group Ⅰ included most of local strains with flowering characteristic, and Group Ⅱ included non-flowering, Wanju and Gunsan strains. Wanju and Gunsan local strains with good growth and non-flowering characteristics were favorable as edible resources. In analysis of genetic relationship among 31 strains by randomly amplified polymorphic DNA (RAPD) with fifteen 10-mers and twelve 20-mers random primers, the result analyzed by 20-mers primers was matched well with morphological characteristics compared to 10-mers primer. Thirty-one strains were clustered into 2 groups. Group included twenty-Ⅰ seven local strains, and Group Ⅱ included Keumsan, Bucheon, Pocheon and Yongin strains with long hight, slender stem, narrow leaf, and abundant flowering characteristics.
○ In vitro multi-shoots were produced on MS medium with 0.2 mg/L BA in 80-90 days after node culture. For In vitro conservation of genetic resources, when shoot tips were cultured on MS media containing 5% sucrose and 1.2% agar, 0.2mg/L BA, and 10-20 mg/L ABA at 4℃, or MS media containing 1.2% agar, 0.2 mg/L BA, and 6-9% sucrose, they could be conserved over one year without subculture. In vitro shoot-tips were successfully cryopreserved in PVS 2 solution with the preculture vitrification technique using 0.4-0.6M sucrose. The range of 11.1-16.7% post-thaw regeneration was obtained. The result showed a low viability, and S. sarmentosum was one of recalcitrant species for cryopreservation. Accordingly, the conservation of genetic resources of S. sarmentosum was more efficiency in vitro conservation using hard agar medium or ABA.
○ The weight loss of fresh shoots showed significant difference (8.7-25.3%) between just before storage and 10 days after storage. Wanju local strain showed the lowest weight loss, and the most severe case was Pohang local strain. The freshness of shoot after storage was maintained in order of Wanju, Gunsan, Wando and Pohang local strain. When the fresh shoot was stored with wrap film-packaged condition, the freshness of shoots was maintained for 8 days after 5℃ storage.
○ Vitamine C content and antioxidant activity of 13 local strains were analysed. Vitamin C contents of 13 local strains were ranged from 407.8 to 434.4 mg/kg, and it was high in Gunsan, Gwangyang, Buan, Wanju, Keumsan, Pocheon, and Bucheon strains. Gwangyang, Suncheon, Wando strains showed high EDA compared to the others. Vitamine C content and antioxidant activity were highest in Gwangyang strain.
2. Investigation of health-benefit function and development of healthy-food with high additive value
A. Improvement of hepatoprotective effect, isolation and identification of cancer inhibitory substance, and development of functional beverage using S. sarmentosum extracts
○ Hepatic protective and cancer inhibitory substance was identified as tetrahydroxyflavonol (quercetin). Hepatoprotective effect to the liver injury induced by carbon tetrachloride in rats was higher in hexane, ethyl acetate, butanol extracts than in silymarine (control).
○ Methanol, hexane, ethyl acetate and butanol extracts was significantly inhibited the induction of fatty liver by ethionine and ethanol, and hexane and ethyl acetate extracts were excellent inhibited effect compared to the others. Inhibitory effect of solvent extracts on human hepatoma HepG2 cell growth was highest in ethyl acetate extract (IC₅₀ = 155.8 μg/mL).
○ After oral adminstration of ethanol and aqueous or solvent extracts of S. sarmentosum in rats, blood ethanol level in aqueous extract was lower 2.3 times than the control, and ethanol oxidation in aqueous extract was more enhanced compared to the solvent extracts. Methanol and butanol extracts were significantly enhanced ADH and ALDH activities of ethanol administered rat.
○ When aqueous or solvent extracts fermented before oral adminstration of ethanol in rats, blood acetaldehyde level in solvent extracts was decreased about 50% after one hour compared to the control, that in aqueous extract was hardly detected after four hour. Blood ethanol and acetaldehyde levels in human were significantly decreased by fermentation of aqueous or solvent extracts before oral adminstration of ethanol. Accordingly, a alcohol detoxification beverage contained S. sarmentosum extracts was prepared, and it is appling the patent.
B. Isolation and identification of antioxidant- and immune-activity substances, mechanisms of antioxidant and immune activities, and development of functional jelly using S. sarmentosum extracts
○ DPPH radical scavenging activity was most superior in subfraction E4 from ethyl acetate among solvent extracts, and the substance was identified as quercetin. Immune activity was most superior in subfraction E5 from ethyl acetate among them, the substance was identified as esculetin of coumarin group.
○ After oral adminstration of S. sarmentosum solvent extracts for 4 weeks in rats, antioxidant activity was similar or high compared to the tocopherol or ascorbic acid (controls). Polyphenol concentration was high in methanol and ethyl acetate extracts, and it accorded that antioxidant activity was high in oral adminstration of methanol extract.
○ When the RAW 264.7, murin macrophage cell line, was cultured in medium adding solvent extracts or LPS (lipopolysaccharide) for 2 days, nitric oxide (NO) production was decreased by methanol, hexane and ethyl acetate extracts in 0.1 μg/mL LPS level. Levels of iNOS protein with LPS treatment was decreased in solvent extract treatments.
○ In rats fed with high fat diet and S. sarmentosum extracts, an aspect of protein expression increased glutathione peroxidase and glutathione reductase related to the antioxidant activity, and COX2 related to the immune activity.
○ Accordingly, the Sedum jelly was prepared using S. sarmentosum juice supplemented with sugar and gelling agent, and contents of Ca, Fe, K, ascorbic acid and organic acids were increased to high levels of S. sarmentosum juice. Feeding preference was highest in jelly contained S. sarmentosum juice than solvent extracts.
C. Isolation and identification of substances related to lipid metabolism and blood flow and immune activities, mechanisms of S. sarmentosum extracts and powder on lipid metabolism and blood flow, and development of functional tea using S. sarmentosum power
○ Inhibition of S. sarmentosum extracts in rat adipocytes proliferation was highest in ethyl acetate subfraction E1, the substance was identified as caffeic acid.
○ IC₅₀ of ethyl acetate extract on inhibition of MC3T3-L1 differentiation was 66.98 μg/mL, and PPAR-gamma gene expression was hardly detected in adding ethyl acetate extract.
○ When S. sarmentosum power (1.5-4.5%) was added in hypercholesterol feed of rat, the level of weight gain in more increased S. sarmentosum power was low in spite of higher feed intake. In feeding S. sarmentosum power, total cholesterol, TG and LDL levels, and atherogenic indices were significantly decreased, and liver lipid and serum ALP levels was low compared to the control. Diet intake of S. sarmentosum could be prevented obesity, non-alcoholic fatty liver disease and hyperlipidemias induced by high fat diet, and it, inhibited hypertension, atherosclerosis and diabetes etc. Accordingly, the Sedum tea was prepared using S. sarmentosum powder.
3. Analysis of inorganic, nutritive and volatile contents and estimation of safety and stability for products of healthy-food
○ Water content was higher at vegetative stage than flowering stage, and the range was 92.8-98.9%. K, Ca and Fe contents were high in Keumsan, Wando, Gunsan strains. Ascorbic acid and beta-carotene was highest in Gunsan strain. Sixteen volatile compounds was analyzed from shoot, and main compound was pyrazine and its derivatives.
○ Beverage, jelly or tea adding solvent extracts, aqueous extract or powder of S. sarmentosum were safe in repeated toxicity test, antimutagenic test, cytotoxicity test, inhibition effect of cancer cell, and hepatotoxicity test. Beverage with hepato-protective activity was stable for 3 months at room temperature, and powder tea was stable after 6 months at room temperature. Jelly adding aqueous extract was stable for 10 days at 4℃.

목차 Contents

• 표지 ... 1
• 제출문 ... 2
• 요약문 ... 3
• SUMMARY ... 16
• CONTENTS ... 21
• 목차 ... 23
• 제 1 장 연구개발과제의 개요 ... 25
• 제 1 절 연구개발의 목적 ... 25
• 제 2 절 연구개발의 필요성 ... 26
• 제 3 절 연구개발의 내용 및 범위 ... 28
• 제 2 장 국내외 기술개발 현황 ... 31
• 제 1 절 국내 돌나물 연구 현황 ... 31
• 제 2 절 국외 돌나물 연구현황 ... 33
• 제 3 장 연구개발 수행 내용 및 결과 ... 34
• 제 1 절 돌나물 유전자원 유연관계 분석, 기내 보존 및 저장성 향상 기술 개발 ... 34
• 1. 서 언 ... 34
• 2. 연구 방법 ... 35
• 3. 결과 및 고찰 ... 41
• 가. 돌나물 유전자원의 유연관계 ... 41
• 나. 돌나물 유전자원 기내 활성보존 및 초저온 동결보존 ... 56
• 다. 수확후 저장온도에 따른 품질변이 ... 62
• 라. 지역 수집종들의 항산화 활성 및 비타민 C 함량 ... 69
• 4. 결 론 ... 72
• 제 2 절 돌나물의 건강기능성 규명 및 고부가가치 제품의 개발 ... 74
• 1. 서 언 ... 74
• 2. 연구방법 ... 75
• 3. 결과 및 고찰 ... 78
• 가. 간기능개선 및 간암억제 물질의 분리·동정과 효과 탐색 및 제품개발 ... 78
• 나. 돌나물의 항산화 및 면역활성물질의 분리·동정과 효과 탐색 및 제품개발 ... 119
• 다. 지질대사 및 혈행개선 물질의 분리·동정과 효과 탐색 및 제품개발 ... 145
• 4. 결 론 ... 166
• 제 3 절 돌나물의 성분분석 및 건강기능성 돌나물제품의 안전성 및 안정성 평가 ... 169
• 1. 서 언 ... 169
• 2. 연구방법 ... 170
• 3. 결과 및 고찰 ... 173
• 가. 돌나물 주요성분 ... 173
• 나. 간기능 개선 및 간암 억제작용이 강화된 돌나물 제품(숙취해소음료)의 안전성 및 안정성 평가 ... 177
• 다. 항산화 및 면역활성이 강화된 돌나물 제품(돌나물 젤리)의 안전성 및 안정성 평가 ... 187
• 라. 지질대사 및 혈행 개선 기능이 강화된 돌나물 제품(돌나물 차)의 안전성 및 안정성 평가 ... 195
• 4. 결 론 ... 201
• 제 4 장 목표 달성도 및 관련분야에의 기여도 ... 203
• 1. 연구 목표 달성도 ... 203
• 2. 주요 연구 성과 및 활용 실적 ... 206
• 제 5 장 연구개발결과의 활용계획 ... 209
• 제 6 장 연구개발 과정에서 수집한 해외 과학기술 정보 ... 211
• 1. 숙취 해소제 시장 동향 (한국/일본/중국/대만) ... 211
• 2. 기타 관련분야의 환경변화 ... 212
• 제 7 장 참고문헌 ... 214
• 끝페이지 ... 221