초록

○ 연구결과 - 염증 개선 효능을 가진 오가피 생잎(분말) 및 추출물(액상)을 이용한 가공품 개발
1. 오가피 잎 추출물 대량생산 공정 확립 및 제품화 (제1세부 및 위탁)
1) 오가피 원료 선정
2) 전처리 공정의 최적화
3) 용매량, 온도, 시간 등 추출 조건 확립
4) 제제화 시험용 시험물질의 생산
5) 시제품의 규격화 및 표준화

Abstract ▼

Ⅳ. Results of the Project and Their Applications
Section 1. Investigation and establishment of large scale -manufacturing process of Acanthopanax leaf extracts and the product development of leaf extracts (liquid form) and dried leaf (powder form) having an improving effect on inflammation (Contr

Ⅳ. Results of the Project and Their Applications
Section 1. Investigation and establishment of large scale -manufacturing process of Acanthopanax leaf extracts and the product development of leaf extracts (liquid form) and dried leaf (powder form) having an improving effect on inflammation (Contract Standardization of Acanthopanax leaf raw materials, the optimization of the condition of a manufacturing process, the establishment of the analyses of approvals and the basis of quality control)
We established simultaneous quantitative analysis for the index components, eleutheroside E and chiisanoside, The quantitative analyses demonstrated that index components were comparatively more detected from 3 kinds of species, Acanthopanax sessijifiorum, Acanthopanax senticosus var.subinermis, Acanthopanax divaricatus among general domestic species.
The extrusion process of the pre-treatment process was established as a new manufacturing process to raise yield of active components. Extrusion process using the mixture of at 9 : 1 ratio of Acanthopanax leaf and corn starch was performed under this conditions barrel temperature 130 'C water content 40%, screw revolution speed 200rpm, material feeding speed 110g/min.
The extraction process was optimized that 10 times water of an extraction solvent volume added to powered extrude and extracted it 11 0 'C for 2 hours.
The test materials were prepared by above optimized extraction process. The liquid form drinks, soy sauce, capsule and tea were manufactured using the test materials and the marker components from the test materials and these approvals were quantitatively analyzed. We confirmed the material stability by checking the marker components.
Section 2, Antioxidant and Anti -inflammatory Effects of Acanthopanax leaf extracts in vitro and in an acute gastritis mouse model, and Preparation of tea with A, senticosus leaves and their sensory characteristics
I} Antioxidant Effects of Acanthopanax leaf extracts Extracts
Acanthopanax senticosus var. subinermis (A. senticosus) has been used in East Asia as a traditional medicine. The purpose of this study is the investigation of an extraction method for A.senticosus leaves to improve their anti -oxidant and anti -inflammatory activities in in vitro and in vivo. In additions, we verified the availability for the A. senticosus-products prepared with developed processes using a sensory evaluation.
Six extracts were prepared: extracted with water (W), ethanol (Eth) , water or ethanol after ultra high pressure (WP, EthP), and water or ethanol after an extrusion process (WEx, EthEx). DPPH radical-scavenging activity and ferric reducing ability showed that all extracts had a significant anti -oxidant activity.
2) Anti -inflammatory Effects of Acanthopanax leaf extracts in RA W264. 7mouse macrophage cell
In anti -inflammatory activities, Eth and EthEx significantly inhibited the release of LPS-induced nitric oxide (NO), tumor necrosis factor-a , and interleukin-1i3 in a concentration-dependent manner independent of cell viability in RAW 264.7 mouse macrophage cells. Inducible NO synthase (iNOS) protein was significantly decreased by the EthEx extract. These findings suggested that the Eth and EthEx extracts of A. senticosus leaves had anti -inflammatory activities. and the EthEx extract. m particular. suppressed LPS-induced NO through the down-regulation of iNOS and decreased the release of pro-inflammatory cytokines.
3) anti - oxidant and anti - inflammatory activity of Acanthopanax leaf extracts in an acute gastritis mouse model,
To investigate whether A. senticosus leaves could exert anti-oxidant and anti -inflammatory activity in VIVO m an acute gastritis mouse model, we measured malondialydehyde (MDA) , NO and CRP levels in bloods after providing orally extrusion -extracts containing high or low concentration of A. senticosus leaves for an acute gastritis mouse model. A. senticosus leaves significantly suppressed the formation of a lipid peroxide, MDA, and an inflammation index, CRP independent of an extrusion process. In particular, an extrusion extract containing a low concentration of the leaves showed a more potential result in a level of blood - NO than the other extracts.
4) Preparation of tea with Acanthopanax leaf extracts and evaluation of their sensory characteristics
A sensory evaluation was conducted to determine effect of extrusion or roasting process on the consumer preference for tea product prepared with A. senticosus leaves. We prepared tea samples with raw leaves, extrusion leaves and extrusion leaves after roasting several times. The results of a sensory evaluation demonstrated that extruded leaves showed a higher score than raw leaves, and roasting process increased preference for taste, color, flavor and overall desirability than non-roasted leaves. Extruded leaves roasted three times showed a significantly higher sensory scores than those of other samples.
Section 3. Alleviation Effect of Acanthopanax leaf extracts on Gastritis.
1) Verification of the Effect In -vitro (valuation of inhibition effect on human mast cell degranulation)
We measured inhibition effect of Acanthopanax leaf extracts which followed six extraction processes each, on human mast cell degranulation in this cooperative study and obtained the following results. Because all extraction processes of the six products showed strong histamine inhibition effect, it can be estimated that Acanthopanacis Folium extracts have an alleviation effect on gastritis. Most results of extrusion extracts especially showed significant data.
Very high pressure extraction showed better data. According to this cooperative study. it seems that extrusion extraction and very high pressure extraction have a comparative advantage. Water and alcohol are available solvents for the extractions and their crystals can be decided by economical efficiency.
2) Verification of the Effect In -vivo (valuation of inflammatory inhibition effect on gastritis induced rats)
We observed effect of Acanthopanax leaf extracts on rat's Compound 48/80 induced acute gastritis through this study. Acanthopanax leaf extruded high-dose treated group (200 mg/kg). Acanthopanax leaf extruded low-dose treated group (40 mg/kg) and Acanthopanax leaf non -extruded treated group (200 mg/kg) were injected into rats. Four days after injections. Compound 48/80 was abdominally injected to rats to induce a gastritis. Three hours after injections. blood was collected and the stomach was extracted to observe change of indexes of such as histamine. MDA. SOD and adherent mucus. High -dose and low-dose group of extruded Acanthopanax leaf extract and non-extruded group showed more relieved sores than control group and average PAS positive reaction.
PAS stammg negative reaction rates on mucosal surface of high -dose group and low-dose group were statistically lower than control group. Non-extruded group's was lower than control group . but there was no statistical significant. Histamine counts in serum showed no statistical difference.
MDA value showed inhibition effect in high-dose group and low-dose group. but there was no statistical difference. There was no inhibition effect in non-extruded group. SOD value showed no effect in high-dose group. but it showed effect in low-dose group and non-extruded group. Low-dose group especially had no difference from normal group and showed the most effective SOD activity promoting value.
XO value showed no effect in high-dose group. but it showed statistically different effect in low-dose group and non-extruded group. High-dose group showed MPO value more decrease than control group. but there was no statistical significant. Also. low-dose group and non-extruded group showed no statistically significant effect. too.
Adherent mucus value showed outstandingly statistically significant effect m high-dose group and non-extruded group. There was effect m low-dose group.
According to the results above. it is concluded that Acanthopanax leaf extracts have alleviation effect that inhibits gastritis from advancing
Section 4. Alleviation Effect of Acanthopanax leaf extracts Extract on Arthritis.
1) Verification of the Effect In-vitro (valuation of differentiation inhibition effect on osteoclast destructing joint )
We measured (by using TRAP staining method, TRAP activity method, NF-kB activation measurement) inhibition effect of Acanthopanacis Folium extracts , which followed six extraction processes, on osteoclast differentiation in this cooperative study and obtained following results, Because all the six products of extraction processes showed strong inhibition effect on osteoclast differentiation, it can be estimated that Acanthopanacis Folium extracts have alleviation effect on arthritis, Most results of extrusion extracts especially showed significant data, in case of hot water extraction showed better data than others, According to this cooperative study, it is concluded that all the six extraction processes are available, Water and alcohol are available solvent for the extractions and their crystals can be decided by economical efficiency,
2) Verification of the Effect In -vivo (valuation of inhibition effect on arthritis induced rats)
We observed effect of Acanthopanacis Folium extracts on rat's Complete Freund's Adjuvant (CFA) induced arthritis through this study, While Acanthopanacis Folium extruded high-dose treated group (200 mg/kg), Acanthopanacis Folium extruded low-dose treated group (40 mg/kg) and Acanthopanacis Folium non-extruded treated group (200 mg/kg) were injected into rats, we observed morphometric change, morphology changes, and cytokine content changes in paw exudates and histopathological change.
Visibly checked grades of arthritis were more relieved than high -dose group, low-dose group and non-extruded group, The body weights were more increased in high-dose group and non-extruded group than in control group. The body weights were statistically more increased in high-dose group on 18th and in non-extruded group on 16th and 20th than in control group.
The paw edema volume were more decreased in high-dose group, low-dose group and non -extruded group than in control group, The paw edema volume was statistically more decreased in high-dose group on 16th and 18th, in low-dose group on 18th and in non-extruded group on 18th than in control group.
The thickness of ankle joint were more decreased in high-dose group, low-dose group and non-extruded group than in control group, The thickness of ankle joint was statistically more decreased in high-dose group on 18th and 20th and in low-dose group on 18th than in control group.
The white blood cell and neutrophil counts were slightly lower in high -dose group, low-dose group and non-extruded group than in control group, The neutrophil Counts were statistically lower in non-extruded group than in control group, TNF-a and IL-1 i3 contents in the paw exudates were slightly lower in high-dose group , low-dose group and non-extruded group than in control group. The histopathological lesion grades of arthritis were slightly lower in high-dose group, low-dose group and non-extruded group than in control group, The histopathological lesion grades of arthritis was statistically lower in non -extruded group than in control group.
According to the results above, it is concluded that Acanthopanacis Folium extracts have inhibiting effect on advancement of rat's CFA induced arthritis a small amount.

목차 Contents

• 제출문 ... 1
• 요약문 ... 2
• SUMMARY ... 25
• 목차 ... 35
• 제1장 연구개발과제의 개요 ... 38
• 제1절 연구개발과제의 목적 ... 38
• 제2절 연구개발 필요성 ... 38
• 제2장 국내외 기술개발 현황 ... 40
• 제3장 연구개발수행 내용 및 결과 ... 41
• I. 연구개발수행 내용 ... 41
• 제1절 오가피 잎 추출물 대량생산 공정 확립 및 제품화(위탁과제 : 오가피 잎 원료, 추출물 및 시제품의 지표 성분의 정성, 정량) ... 41
• 1. 오가피 원료 선정을 위한 오가피 종류별 잎 원료의 유효성분 정량분석 ... 41
• 2. 전처리 공정의 최적화 ... 43
• 3. 용매량, 온도, 시간 등 추출 조건 확립을 위한 유효성분 정량분석 ... 44
• 4. 제제화 시험용 시험물질의 생산 ... 44
• 5. 시제품의 규격화 및 표준화 ... 44
• 제2절 오가피잎 추출물의 항염 /항산화 효능 및 식품학적 품질평가 ... 45
• 1. 항산화 활성 평가 및 대식세포 (Raw 264.7 cells)의 염증 지표 변화 ... 45
• 2. 위염/관절염 유발 동물 모델을 이용한 염증 지표와 항산화 활성 평가 및 시제품의 소비자 선호도 조사 ... 46
• 제3절 오가피잎 추출물의 위염 개선 효과 ... 49
• 1. In-vitro 에서의 효능 검증 (사람비만세포 탈과립 억제효능 평가) ... 49
• 2. In-vivo 에서의 효능 검증 (위염 유도 흰쥐에 대한 염증억제 효능 평가) ... 50
• 제4절 오가피잎 추출물의 관절염 개선 효과 ... 54
• 1. In-vitro 에서의 효능 검증 (관절파괴를 하는 파골세포 분화억제효능 평가) ... 54
• 2. In-vivo 에서의 효능 검증 (관절염 흰쥐에 대한 억제효능 평가) ... 55
• II. 연구개발 수행 결과 ... 59
• 제1절 오가피 잎 추출물 대량생산 공정 확립 및 제품화(위탁과제 : 오가피 잎 원료, 추출물 및 시제품의 지표 성분의 정성, 정량) ... 59
• 1. 오가피 원료 선정을 위한 오가피 종류별 잎 원료의 유효성분 정량분석 ... 59
• 2. 전처리 공정의 최적화 ... 60
• 3. 용매량, 온도, 시간 등 추출 조건 확립을 위한 유효성분 정량분석 ... 63
• 4. 제제화 시험용 시험물질의 생산 ... 64
• 5. 시제품의 규격화 및 표준화 ... 65
• 6. 요약 및 결론 ... 67
• 제2절 오가피잎 추출물의 항염 /항산화 효능 및 식품학적 품질평가 ... 69
• 1. 항산화 활성 평가 및 대식세포 (Raw 264.7 cells)의 염증 지표 변화 ... 69
• 2. 위염/관절염 유발 동물 모델을 이용한 염증 지표와 항산화 활성 평가 ... 73
• 3.시제품의 관능 평가와 순위 검사 ... 77
• 4. 요약 및 결론 ... 78
• 제3절 오가피잎 추출물의 위염 개선 효과 ... 80
• 1. In-vitro 에서의 효능 검증 (사람비만세포 탈과립 억제효능 평가) ... 80
• 2. In-vivo 에서의 효능 검증 (위염 유도 흰쥐에 대한 염증억제효능 평가) ... 85
• 3. 요약 및 결론 ... 95
• 제4절 오가피잎 추출물의 관절염 개선 효과 ... 97
• 1. In-vitro 에서의 효능 검증 (관절파괴를 하는 파골세포 분화억제효능 평가) ... 97
• 2. In-vivo 에서의 효능 검증 (관절염 흰쥐에 대한 억제효능 평가) ... 103
• 3. 요약 및 결론 ... 116
• 제4장 목표달성도 및 관련분야에의 기여도 ... 118
• 1. 연도별 연구개발의 목표 및 내용 ... 118
• 2. 평가의 착안점 및 기준 ... 120
• 제5장 연구개발결과의 활용 계획 ... 121
• 제6장 연구개발과정에서 수집한 해외과학기술정보 ... 122
• 제7장 참고문헌 ... 123
• 끝페이지 ... 127