초록 ▼

<배경> 와송은 우리나라에서 오래전부터 민간요법으로 간염, 종기에 대한 면역작용, 지혈제 및 암치료제 등으로 사용되어져 왔으며, 최근 와송에 존재하는 phytochemicals로 sterol, triterpenoid류, 플라보노이드류 및 페놀 화합물 등이 분리되었고, 소화기 계통의 암에 효과가 좋은 것으로 알려지면서 이에 대한 연구가 활발히 진행되고 있는 실정이나, 생리활성 물질의 분리나 동물실험을 통한 생물활성 검증 및 와송과 관련된 제품의 개발은 미비한 실정이다.
<내용> 와송의 유용물질을 분리하여 항산화 활성 측정, 실험

<배경> 와송은 우리나라에서 오래전부터 민간요법으로 간염, 종기에 대한 면역작용, 지혈제 및 암치료제 등으로 사용되어져 왔으며, 최근 와송에 존재하는 phytochemicals로 sterol, triterpenoid류, 플라보노이드류 및 페놀 화합물 등이 분리되었고, 소화기 계통의 암에 효과가 좋은 것으로 알려지면서 이에 대한 연구가 활발히 진행되고 있는 실정이나, 생리활성 물질의 분리나 동물실험을 통한 생물활성 검증 및 와송과 관련된 제품의 개발은 미비한 실정이다.
<내용> 와송의 유용물질을 분리하여 항산화 활성 측정, 실험동물을 이용한 항당뇨 활성 및 항암․면역 활성을 실험하여 천연소재인 생약재와 복합조성물을 만들어 기능성을 향상시킨 와송 가공품의 개발과 상품화를 위한 제조공정을 개발하였다.
<성과> 와송의 항산화 및 항당뇨 활성을 확인하였으며, 국내 전문학술지 발표 6 건과 특허 출원을 수행하였으며, 이러한 결과를 이용하여 제품의 개발과 상품화를 통하여 향후 와송 재배농가의 소득 증대와 기능성 상품 매출액의 증대에 기여할 것으로 예상된다.

Abstract ▼

Wa-song(Orostachys japonicus A. Berger), in Korea, is a perennial herb known as a medicinal plant. It has been used as an anti-inflammatory agent for the treatment of hepatitis, boils and piles, and as a hemostatic agent for the treatment of hematemesis, rhinorrhagia and bloody excrement. Its major

Wa-song(Orostachys japonicus A. Berger), in Korea, is a perennial herb known as a medicinal plant. It has been used as an anti-inflammatory agent for the treatment of hepatitis, boils and piles, and as a hemostatic agent for the treatment of hematemesis, rhinorrhagia and bloody excrement. Its major bioactive compound, gallic acid (GA), significantly enhanced hepatic ethanol metabolism.
Several phytochemicals, including triterpenes, sterols and flavonoids have been isolated from the plant. Some biological activities such as antioxidant, anti-complementary and cytotoxic have been reported by many authors in Korean. Furthermore, methanol extract of Wa-song also been shown to have a protective effect against H2O2-induced apoptosis in mouse hypothalamic neuronal cell. The active ingredients of Wa-song protecting neuronal cell from reactive oxygen species were proposed to be hydrophobic compounds. Water extract of the aerial part of Wa-song also been shown to have an inhibitory activity against HIV-1 protease.
Antioxidant activity of Wa-song was analyzed to clarify the influence of extractive solvent and drying method such as sun, hot-air and freeze drying. The contents of total phenols and flavonoids were higher in 95% ethanol extracts than water extracts, significantly. Ability of reducing power and DPPH radical, hydroxyl radical and nitrite scavenging ability were higher in the hot-air dring sample > freeze drng sample > sun dring sample, in order and their abilities were also higher in 95% ethanol extracts than water extracts. In conclusion of this experiment, antioxidant activities of Wa-song extracts were in proportion to the contents of total phenols and flavonoids. And, hot-air drying for enhancement of antioxidant activity of Wa-song is better than freeze and sun drying samples. In reaction system containing linoleic acid, the antioxidant activities against lipid oxidation enhanced in proportion to storage time. The antioxidant activity of ethanol extracts was higher than that of water extracts. In the drying methods, Wa-song extracts showed higher antioxidant activity in the order of hot-air, freeze and sun dring. The lipid oxidation the highest antioxidant activity, in system containing the promoting factors, such as Fe⁺² and Cu⁺² ions, also showed that in ethanol extract of hot-air dring sample. The antioxidant activity of medicinal plants and Wa-song composites(MW) was analyzed. DPPH and ABTs radical scavenging ability were distinctly appeared in the low concentration of medicinal plants and Wa-song composites. The inhibitory activity of MW against α-glucosidase showed significantly increased by added amount of Wa-song.
The antioxidant activity of Wa-song was tested in vitro system containing oil and lard. Three kind samples of Wa-song was prepared water of hot-air drying(HW), ethanol extracts of hot-air drying(HE) and water extracts of freeze drying(FW). The different levels (0.1, 0.5 and 1.0 g/100 g) of HW, HE and FW were added to soybean oil and lard. Chromaticity, anisidine value, acid value, peroxide value(POV) and TBA value in oils were measured periodically during their storage for 28 days at 60℃ and 48 hrs at 180℃. The chromaticity of edible oils was generally increased with prolonged storage days and increase in quantities of extracts at 60℃. The anisidine value showed not significantly increased during storage for 14 days. In soybean and lard added HE, its value was lower than control and BHT for storage for 28 days at 60℃. The acid value of HW was significantly increased during storage from 14 to 21 days in soybean oil and from 7 to 14 days in lard. POV was highly increased between storage for 7 and 14 days in reaction system of soybean oil. TBA values of all samples were lower than control and 0.02% BHT during their storage.
During the storage at 180℃, the chromaticity of edible oils was generally increased with prolonged heating time and HW was higher than FW. The anisidine value showed significantly increased during heating for 48 hrs. The acid value was not significant by added amount of Wa-song. Its value in HW added sample was lower than FW, after heating for 48 hrs. POV was lower HW than FW, also. After heating for 24 hrs, TBA values in soybean oil containing HW and FW added sample was lower than control. In lard, its value in HW and FW added sample was lower than control during heating for 12~48 hrs. In adding HE, it was significantly decreased by sample concentration after storage 36 hours at 180℃ storage.
The antidiabetic potential of Wa-song in streptozotocin induced diabetic rats was conducted. Male Sprague-Dawley rats were divided into 8 groups at different dose of 0.1% or 0.5% were as the diabetic experimental groups. The long time administration of extracts from Wa-song decreased the fasting blood glucose levels and serum glucose level in diabetic rats significantly decreased from the second week, and especially rats in the group fed with FW at dose of 0.5% showed the more significant effect comparing with the diabetic control group fed with basal diet. The significant decrease of serum low density lipoprotein cholesterol(LDL-C) levels in diabetic rats could reduce the risk of cardiovascular disease. There was no significant increase in hepatic glycogen levels of rats, but hepatic protein levels were increased significantly in rats of the group fed with HE at dose of 0.5% and FW at dose of 0.1%. From the results of hepatic TBARS levels, CD levels and hepatic antioxidant activity of rats, they demonstrated that antioxidant and anti-lipidperoxidant ability of diabetic rats were improved.
Effects of feeding diets containing medicinal plants and Wa-song composites(MW) in streptozotocin induced diabetic rats was examined. Groups were divied into MW-0(medicinal plants: Wa-song=1:0), MW-1(medicinal plants: Wa-song=1:1) and MW-3(medicinal plants: Wa-song=1:3) in induced diabetic rats. Oral glucose tolerant test(OGTT) was significantly decreased by time, group fed MW-3 shown 202.00 mg/dL after 180 min. And its level was same as the fasting blood glucose level.
Blood glucose and glucosylated hemoglobin was significantly decreased by added amount of Wa-song. Glycogen content of liver was significantly increased by Wa-song contents.
Wa-song was activated macrophage and than elevated nitric oxide(NO) concentration. Secreation of cytokine was activated anti-tumor and immunostimulation. Anticancer activity of Wa-song was significantly decreased on HT-29 and HepG-2 cells. In MTT assay, growth inhibitory effects on AGS, HT-29 cells were significantly increased by adding amount of Wa-song. Anticancer activity of Sang-hwang mushroom extract and Wa-song composites(1:2 or 2:1/v:v) on MCF-7 cells was 56.55±2.18%, 73.59±5.33%. And its levels were significantly increased than control.
Package design and experimental products in a liquids, pills, tabelts containing Wa-song were developed. In the organoleptic characteristics of products, the overall acceptability was relatively higher.

목차 Contents

• 표지 ... 1
• 제출문 ... 2
• 요약문 ... 3
• SUMMARY ... 10
• CONTENTS ... 13
• 목차 ... 15
• 제1장 연구개발과제의 개요 ... 18
• 제2장 국내외 기술개발 현황 ... 20
• 제3장 연구개발수행 내용 및 결과 ... 21
• 제1절 와송의 생리활성 및 상품화 ... 21
• 1. 서 론 ... 21
• 2. 와송 유용물질의 분리 및 분석 ... 22
• 가. 채취시기별 물질 분리 ... 22
• 나. 용매별 물질 분리 ... 22
• 다. 와송의 유용물질 분리 ... 24
• 3. 와송 천연물소재를 이용한 최적조성물 개발 ... 27
• 가. 항당뇨 활성 부원료 선정 ... 28
• 나. 와송 및 선정생약재의 추출수율 ... 34
• 다. 와송 최적조성물의 배합조건 ... 35
• 4. 와송 및 최적조성물의 항산화활성 ... 36
• 제2절 와송의 가공특성 및 생산공정 구축 ... 39
• 1. 와송의 가공특성 ... 39
• 가. 건조방법에 따른 건조수율 ... 40
• 나. 건조방법을 달리한 와송의 물추출물 제조 ... 41
• 다. 건조방법을 달리한 와송의 에탄올추출물 제조 ... 42
• 2. 와송의 유용물질을 함유하는 조성물을 이용한 제형화 ... 44
• 가. 바이오헬스 소재 및 기능성식품의 형태 ... 44
• 나. 과립분말형 시제품 제형화 공정 ... 45
• 다. 액상형 시제품 제형화 공정 ... 46
• 라. 와송을 활용한 환형, 파우치형 및 정제형 제품 제형화 공정 ... 46
• 마. 대량생산시스템 설계 ... 48
• 3. 와송 천연물소재 및 최적조성물 제조공정 ... 49
• 제3절 와송의 항산화 활성 구명 ... 51
• 1. 서 론 ... 51
• 2. 재료 및 방법 ... 52
• 가. 항산화 활성 측정 ... 52
• 나. 식용유지기질에서 항산화 활성 측정 ... 55
• 3. 결과 및 고찰 ... 56
• 가. 와송 채취시기별 항산화 활성 ... 56
• 나. 용매별 추출물의 항산화 활성 ... 69
• 다. 처리조건별 와송 추출물의 항산화 활성 ... 81
• 라. 식용유지 기질에서 와송 추출물의 항산화 활성 ... 90
• 마. 생약재복합물 및 와송 추출물의 항산화 활성 ... 111
• 제4절 와송의 항당뇨 활성 구명 ... 121
• 1. 서 론 ... 121
• 2. 재료 및 방법 ... 122
• 가. 실험동물, 사육조건 및 식이조성 ... 122
• 나. 당뇨 유발 ... 122
• 다. 식이섭취량, 식이효율 및 체중측정 ... 122
• 라. 경구 당부하 검사 ... 123
• 마. 실험동물의 처리 ... 124
• 바. 혈액 분석 ... 124
• 사. 조직성분 분석 ... 125
• 아. 통계처리 ... 127
• 3. 결과 및 고찰 ... 127
• 가. 와송 추출물의 항당뇨 활성 ... 127
• 나. 생약재복합물과 와송 추출물의 급이에 따른 항당뇨 활성 ... 141
• 제5절 와송의 항암.면역 활성 구명 ... 158
• 1. 서 론 ... 158
• 2. 재료 및 방법 ... 159
• 가. 세포 주 배양법 ... 159
• 나. 일산화 질소 측정법 ... 160
• 다. Cytotoxicity 측정법 ... 160
• 라. Sulforhodamine B (SRB) assay ... 160
• 마. Cell morphology and nuclear staining ... 161
• 바. Flow cytometric analysis of cell cycle ... 161
• 사. MTT assay ... 161
• 3. 결과 및 고찰 ... 162
• 가. 와송 추출물에 의한 NO 생산 ... 162
• 나. 와송 추출물에 의한 대식세포의 cytotoxicity 생산 ... 166
• 다. 와송 추출물의 암세포 성장 억제 효과 ... 167
• 라. 와송 추출물의 DNA fragmentation ... 170
• 마. 와송 추출물의 FACS에 의한 세포주기 변화 ... 171
• 바. 와송 추출물 및 생약재 혼합 조성물의 암세포 성장 억제능 ... 172
• 사. 와송과 상황버섯 추출물 및 혼합 조성물의 암세포 성장 억제능 ... 175
• 제6절 시제품 개발 및 상품화 ... 180
• 1. 제품별 포장디자인 개발 ... 180
• 가. 와송 액상제품 포장디자인 ... 180
• 나. 환제품 및 정제형 제품의 포장디자인 ... 180
• 다. 액상 시제품의 품질평가 ... 181
• 라. 개발시제품의 관능평가 ... 182
• 2. 건강기능식품 제조방법 및 규격 ... 185
• 3. 특허출원 ... 187
• 4. 제품 홍보 ... 201
• 제4장 목표달성도 및 관련분야에의 기여도 ... 202
• - 1차년도 ... 202
• - 2차년도 ... 203
• - 3차년도 ... 204
• 제5장 연구개발 성과 및 성과활용 계획 ... 205
• 제1절 연구개발결과의 활용방안 ... 205
• 제2절 기대성과 ... 205
• 1. 기술적 측면 ... 205
• 2. 경제적ㆍ산업적 측면 ... 206
• 3. 연구성과 ... 206
• 제6장 연구개발과정에서 수집한 해외과학기술정보 ... 208
• 제7장 참고문헌 ... 209
• 끝페이지 ... 214