보고서 정보
주관연구기관 |
도울농산(농) |
보고서유형 | 최종보고서 |
발행국가 | 대한민국 |
언어 |
한국어
|
발행년월 | 2011-12 |
과제시작연도 |
2010 |
주관부처 |
농림축산식품부 Ministry of Agriculture, Food and Rural Affairs(MAFRA) |
등록번호 |
TRKO201400026349 |
과제고유번호 |
1545002463 |
사업명 |
농림기술개발 |
DB 구축일자 |
2014-11-29
|
DOI |
https://doi.org/10.23000/TRKO201400026349 |
초록
○ 연구결과
- 마늘유래 유황함유 성분중 기능성 불회발성 성분의 분리정제 및 분석법 확립
- 마늘시료의 혈행개선(혈소판 응집억제활성)을 숙성 흑마늘이 농도의존적으로 억제함
- 인체 임상 실험을 통한 평가에서 흑마늘 추출액 섭취군이 placebo 보다 혈소판 응집능이 증가함
- 흑마늘 숙성공정 표준화 및 기능성 유황성분의 바이오마크 함유식품 개발
Abstract
▼
IV. Research Results and Application
1. Research results
1) Establshment of isolation and selection of biomarker materials of garlic origin for improvement of blood flow
① Isolation and development of analytical methods of garlic-specific sulfur compounds like alliin, isoalliin, methiin,
IV. Research Results and Application
1. Research results
1) Establshment of isolation and selection of biomarker materials of garlic origin for improvement of blood flow
① Isolation and development of analytical methods of garlic-specific sulfur compounds like alliin, isoalliin, methiin, and three gamma-glutamyl pep tides. All six garlic specific sulfur compounds (alliin, isoalliin, methiin, three gamma-glutamyl peptides) were isolated and purified. Analytical methods for the six sulfur compounds were successfully developed. The result was published in SCIE journal in 201l.
② Establishment of analytical method for dialk(en)yl sulfides formed from alk(en)yl thiosulfinates was performed. Allicin has been known for anticancer activity, antithrombotic activity, blood flow enhancing activity, etc. Thiosulfinates decompose and form dialk(en) yl suifides including diallyl disulfides, trisulfide, tetrasulfide, pentasuifide, which the major components of garlic oil. The individual sulfides have been isolated and purified by using recycling pre-parative HPLC. Individual compounds were provided to Professor Lee (#2 subteam) for the study of enhancement of blood flow,
③ Development of analytical methods of S-allyl-L~cysteine (SAC) and S-allylmercapto-L-cysteine (SAMC) from aged black garlic were successfully performed. SAC was analyzed whenever the principal research team leader asked for. SAMC was synthesized and provided to Dr. Ryu and Or. Lee for the study of its efficiency. Analytical methods for SAC and SAMC were firmly established and many samples of fresh garlic and processed garlic were analyzed on the request of Dr. Ryu.
④ General non-expert public expects that garlic has high functional capacities even though enough research has not performed. It is necessary to set directions for useful and effective research on garlic functionality.
2) Functional assay and standardization of garlic products
① To examine the effect of fractions of AGE on platelet aggregation, we treated freshly isolated rat platelets with various fraction of AGE for 5 min and then initiated platelet aggregation with thrombin. AGE showed inhibitory effect against thrombin induced platelet aggregation in a concentration dependent manner and achieved significance between 1-10% (v:v). Notably. S-allylcysteine (SAC). major sulfur compound. showed very weak inhibitory effect against thrombin induced platelet aggregation. But. water soluble fraction and oil fraction of AGE showed the strongest inhibitory effect against thrombin-induced platelet aggregation than other extraction conditions. Oil fraction of AGE inhibited thrombin-induced platelet aggregation significantly in a concentration-dependent manner.
② To exam ine the effect of fractions of AGE on blood coagulation. we treated freshly isolated rat blood plasma with various fraction of AGE for 5 min and then measured prothrombin time and activated partial thromboplastin time. AGE showed no inhibitory effect against blood coagulation.
③ For the identification of active components for antiplatelet effects of AGE. individual components of AGE (garlic oil. SAC, SAMC, DAS, DADS, DATS) were tested for their effects on thrombin induced platelet aggregation. Total percentage aggregation of platelet was reduced concentration-dependent manner. Garlic oil and OATS showed that the significant inhibitory effect on platelet aggregation than DAS. DADS. Water soluble sulfur compound. S-allylmerceptocysteine (SMAC) showed that the inhibitory effect on platelet aggregation than followed by SAC. Also, individual components of AGE were tested for their effects on ADP or collagen induced platelet aggregation. SAMC, garlic oil and three dially sulphides inhibited ADP induced platelet aggregation in a dose-dependent manner. And garlic oil and three dially sulphides inhibited collagen induced platelet aggregation. As a result, water soluble sulfur compound, SMAC and three dially sulphides showed the most potent inhibitory effect on agonists-induced platelet aggregation.
④ To confirm the anti platelet effect of SAMC and gad ic oil, we conducted an in vivo platelet aggregation measurement after single or multiple oral administration of SAMC (1500 g/kg body weight once daily) and garlic oil (1ml or 3ml/kg body weight once daily) as control corn oil. As a result. thrombin and ADP-induced platelet aggregation was significantly inhibited by SAMC and garlic oil. However, treatment of SAMC and garlic oil not inhibited food intake and body weight.
⑤ We have demonstrated that AGE and its compoenents inhibited agonists-induced platelet aggregation and activation, leading to reduced thrombus formation" In addition, repeated oral administration of SAMC and garlic oil manifested more potent antiplatelet aggregation effects in vivo, indicating that long-term intake of SAMC and garlic oil could be more effective in the prevention of thrombotic events. More importantly, this study has provided, we believe, important evidence for the utility of the common food materials for the effective prevention of cardiovascular diseases.
3) Development of the highly functional garlic products. For export items
① We were examined t he healthy functional properties unique aging garlic. T he majority of aged black garlic is the bulk of dry weight is composed of fructose-containing carbohydrates, followed by protein, fatty acids, unvolatile organic acids and free amino acids. the chemical composition, organoleptic test. and improvement of lipid, metabolism antithrombogenic capacity were examined. Effects of its extract were compared to those of raw garlic extracts. T he color belt, smell, taste garlic's color are rendered black by spontaneous aging on 15 days. under the conditions of high temperature (70-80℃) and high humidity (90-95%, RH) according to aging periods of raw garlic.
② Aged black garlic also contains moderate levels of ,B-carotene and α-tocopherol. It also has increased a high total phenolic, f1avonoids and antioxidant status following aged periods.
③ Total phenols, f1avonoids, and S - allylcystein(SAC), S - alIyl- mercaptocysteine(SAMC) contents were the highest in 15 days aged black garlic. Aged black garlic have higher level of electronic donating abilities(EDA) than that of raw garlic. In terms of its superoxide dismutase(SOD) activities, the aged black garlic of 15 days showed higher activity as compared to the raw garlic.
④ In order to conduct the effect of raw garlic and 10, 15 or 20 days aged black garlic on lipid metabolism, antioxidative capacity in rats , Sprague-Dawley male rats weighing 220± 20 g were blocked in to seven group according to containing 5% of raw garlic or aged black garlic at different aged periods of 10, 15 or 20 days. Especially 15 days is lower plasma total lipids, triglyceride and total cholesterol LDL concentration. HDL cholesterol increased than that of other samples garlic.
⑤ We investigated the aged garlic extracts and garlic oil for pre-c1inical test on platelet aggregation. The aged garlic extracts and garlic oil showed inhibitory effect against collagen/ADP and collagen/EPI induced platelet aggregation in pre-clinical human test.
⑥ The extraction of garlic oil using supercritical carbon dioxide was performed in a semi-continuous flow extractor at the optimal conditions of pressure and teamperature 268-270 bar. and 57-61℃. The most efficiency of extraction of garlic was achieved the highest yield of allicin and other orgamosulfur compounds. Especially, extraction of thiosulfinate are depended on extraction. In conclusion, aged black garlic has a potential to be used as a functional health food with benefit effects on lowering cholesterol and antithromotic levels.
4) Verification of t he effectiveness of garlic by clinical trial
① Subject enrollment
Present study investigated the effect of aged garlic extract
on blood flow in healthy adults using a 12-week, randomized, double-blind, placebo-controlled clinical trial. Sixty subjects were randomly assigned to two treatment groups and were intake to aged garlic extract or placebo during the 12-weeks.
Sixty subjects were randomly divided into aged garlic (n=30) or placebo (n=30) group. Fifty-eight subjects finished the study according to protocol and one subject was withdrawn. One subject is ongoing and will be ended according to the schedule.
② Assessing the Primary efficacy
After the 12 weeks of supplementation, epinephrine and ADP induced platelet aggregation are increased. But there was no significant difference between aged garlic and placebo groups.
③ Assessing the Secondary efficacy
After the 12 weeks of supplementation, systolic and diastolic blood viscosity was no significant difference between aged garlic and placebo groups.
④ Assessing the Safety
After the 12 weeks of supplementation, heart rate in the aged garlic group was significantly reduced compared to the placebo group. Also there was no clinically significant change of safety parameter.
⑤ Conclusion
In conclusion, after the aged garlic supplementation, there was no significant change for blood flow. But heart rate significantly decreased in aged garlic group. So, these results suggested that aged garlic extract supplementation may have positive effect on human health.
2. Application
1) Large scale of aged black garlic processing for lowering cholesterol or improvement of blood circulation derived garlic and standardization of items quality.
2) Selection of biomarker containing lowering cholesterol and improvement of blood circulation derived garlic and development of highly valuable garlic items.
3) Confirmation of highly functional compounds derived garlic through pre-c1 inical experiment and practical use for recongnition of functional health food from Korean FDA.
4) Practical use for recongnition of functional health food of garlic through pre-clinical experiment of human.
5) Development of biomedicine containing functional organic sulfur compounds obtained from garlic.
6) Commercial use of useful compound obtained from garlic.
7) Development and commercial use for export items using highly functional garlic.
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