본 연구에서는 시엽의 기능성 화장품 소재로서의 활용 가능성을 평가하기 위해 항산화와 인 간섬유아세포인 CCD-986sk 세포에서의 항주름 효과를 알아보았다. 전자공여능 측정, ABTS+ radical 소거능 측정을 통해 항산화 활성을 확인한 결과, 시엽은 1,000 μg/ml 농도에서 대조군인 ascorbic acid 와 농도 의존적으로 유사한 항산화 활성을 보였다. Elastase 저해 활성 측정, Collagenase 저해 활성 측 정을 통해 항주름 효과를 확인한 결과, 시엽은 1,000 μg/ml 농도에서 대조군인 epigallocatechin gallate 와 농도 의존적으로 유사한 항주름 효과를 나타내었다. UVB로 유도된 CCD-986sk 세포 내 pro-collagen type I의 합성률과 MMP-1의 저해률을 측정한 결과 대조군인 EGCG는 20 μg/ml에서 90.2%, 시엽은 30 μg/ml에서 88.5%의 pro-collagen 합성률을 보였다. 또한 EGCG 20 μg/ml와 시엽 30 μg/ml에서 각각 33.0%, 40.8%의 MMP-1 저해율을 확인하였다. Western blot을 통하여 시엽의 pro-collagen type I과 MMP-1의 단백질 발현량을 측정한 결과 UVB 단독군 대비 시엽을 함께 처리했 을 때 pro-collagen type I의 단백질 발현량이 증가하고 MMP-1의 단백질 발현량이 감소하는 것을 확 인하였다. 위 실험결과에 따라 시엽이 UVB 자극에 의한 광노화를 예방하고 항산화, 항주름 효과가 있음 을 확인하여 화장품의 천연물 소재로서의 이용이 기대된다.
In order to develop new physiologically active polysaccharides from persimmon leaves, two different crude polysaccharides were prepared using hot water (PLW-0) and pectinase digestion (PLE-0) and their immuno-stimulating activities were estimated. PLW-0 and PLE-0 showed similar sugar compositions with 15 different sugars, including rarely observed sugars in general polysaccharides such as 2-O-methyl-fucose, 2-O-methyl-xylose, apiose, aceric acid, 3 deoxy-D-manno-2-octulosonic acid, and 3-deoxy-D-lyxo-2-heptulosaric acid, but the uronic acid content of PLE-0 was lower than that of PLW-0 caused by pectinase treatment. Both PLW-0 and PLE-0 showed potent anti-complementary activity in a dose dependent manner which was similar to a known immuno-stimulating polysaccharide, PSK, from Coriolus versicolor. The activity of PLE-0 at a low concentration (100 ㎍/㎖) was higher than that of PLW-0. In an in vitro cytotoxicity analysis, PLW-0 and PLE-0 (up to 1, 000 ㎍/㎖) did not affect the growth of peritoneal macrophages and Colon 26-M3.1 carcinoma cells. In contrast, they enhanced lymphocyte proliferation activity. Peritoneal macrophages stimulated with PLW-0 and PLE-0 produced various cytokines, such as IL-6 and IL-12. However, PLE-0 was more effective on the cytokine production. Intravenous administration of PLW-0 and PLE-0 significantly augmented natural killer (NK) cell cytotoxicity against Yac-1 tumor cells 3 days after the treatment of polysaccharide fractions. But NK cells obtained from the PLE-treated group showed higher tumoricidal activity even at a low dose of 40 ㎍/mouse. In experimental lung metastasis of Colon 26-M3.1 carcinoma cells, prophylactic administration of PLW-0 and PLE-0 significantly inhibited lung metastasis in a dose-dependent manner and PLE-0 was more effective on the inhibition of cancer metasasis. The results lead us to conclude that the pectinase treated process is indispensable to preparing polysaccharides with higher immune-stimulating activity from persimmon leaves.
The lyophilization of the solution extracted from 60 percent of acetone applied to persimmon leaves, the compounding process in accordance with the solution's concentration, and the gel filteration through Sephadex G-50 of biologically activated substances obstructing enzyme activity, such as tyrosinase, xanthine oxidase, and angiotesin converting enzyme (ACE) led to the assumption that polyphenol was the compound serving as biologically activated substances obstructing enzyme activity. Xanthine oxidase involved in pruine metabolism oxidizes hypoxanthine to xanthine and xanthine to uric acid. In the continuous study for natural compound, nine flavan-3-ols have been isolated from the persimmon leaves. The structures of (+)-catechin, (+)-gallocatechin, procyanidin B-1, pyrocyanidin C-1, prodelphinidin B-3, gallocatechin-(4α→8)-catechin, procyanidin B-7-3-O-gallate, procyanidin C-1-3'-3'-3'-O-trigallate and (-)-epigallocatechin-(4α→8)-epigallocatechin-(4α→8)-catechin were established by NMR and their inhibitory effect on xanthine oxidase activity was investigated. Procyanidin B-7-3-O-gallate, (-)-epigallocatechin-(4α→8)-epigallocatechin-(4α→8)-catechin and procyanidin C-1-3'-3'-3'-O-trigallate showed 94%, 90.69%, 80.90% inhibition at 100 (μ)M and inhibited on the angiotensin converting enzyme respectively. Procyanidin B-7-3-O-gallate and procyanidin C-1-3'-3'-3'-O-trigallate showed 66%, 63% inhibition at 100 (μ)M and inhibited on the xanthine oxidase competitively. Procyanidin C-1-3'-3'-3'-O-trigallate showed 70% inhibition at 100 (μ)M and inhibited on the tyrosinase competitively.
This study was carried out to separate and identify the antioxidative substances in persimmon leaves. The antioxidative substances in persimmon leaves were extracted by methanol. The extract was fractionnated by SEP-PAK cartridge colum. From these results five fractions(F-I~V) were obtained. Antioxidative activity of each fractions was examined by the DPPH methord. The F-II, III and IV showed antioxidative activity and among them F-II and F-III showed the strongest. Five frations were separated by TLC using ethylacetate : chloroform : formic acid : H2O(8 : 1 : 1 : 1 v /v) as the solvent. From these results were obtained spots of Rf 0.71, 0.35 and 0.25. This spots were scraped from the plate and extracted by methanol. The extracts thuse obtained were used for examination of identify by TLC, UV /VIS-spectrophotometer and HPLC. Among them spot of Rf 0.71 were demonstrated to catechin and the spots of Rf 0.35 and 0.25 was suggested to polyphenol substances.
Persimmon is a plant that is native to the Republic of Korea. It leaves are rich in antioxidants and minerals. This study set out to investigate the moisture, color, vitamin C, and mineral contents of five of the most common Korean persimmon leaves (‘Sangju-dungsi’, ‘Sangam-dungsi’, ‘Cheongdobansi’, ‘Gabjubaekmok,’ and ‘Suhong’), and how they varied with the applied drying technique (freeze-drying or hot-air drying at 100℃ for 30 min) and harvesting time (late May or late June). Persimmon leaves were first collected, and then blanched and dried. Our results revealed that freeze-drying was the most effective method in terms of the mineral content, while no significant difference in the vitamin C content was observed regardless of the drying method. The concentration of Mg, Ca, K, and Mn increased from May to June, while the opposite trend was observed for Na, Fe, Cu, and Zn. Persimmon leaves collected in late May had a higher vitamin C content than those harvested in late June. Ultimately, the ‘Sangam-dungsi’ persimmon leaf was found to have the highest mineral content, whereas ‘Gabjubaekmok’ leaves were richer in color (lightness, redness, and yellowness) and had the highest vitamin C content among the analyzed cultivars.
Total phenolic compounds and antioxidant activity were investigated in leaves of organic apricot, filbert, mulberry, persimmon, and pomegranate trees during growing season. Total phenolic compounds and antioxidant activity in leaves of organic apricot and filbert trees were gradually increased from May to June, attained peak in July, thereafter decreased to minimum level in October. In leaves of organic mulberry tree total phenolic compounds and antioxidant activity were simultaneously increased from May to June and sharply decrease to very low level. The leaves of organic persimmon trees showed very high level of total phenolic compounds and antioxidant activity from May to June, thereafter gradually decrease to October. The leaves of organic pomegranate trees showed extraordinarily highest level of total phenolic compounds among five fruit trees investigated in the study during growing stages, as well as maintained higher than 91 percent of antioxidant activity from May to October. Correlation coefficients between total phenolic compound and antioxidant activity of the leaves of five organic trees were the highest in mulberry leaves, and then persimmon, filbert, apricot, and pomegranate, respectively. However, correlation coefficients between total phenolic compound and antioxidant activity of the leaves of organic pomegranate were very low level, and not significant in their relationship.
환경오염원이면서 알레르기성 접촉피부염 유발제인 NiSO₄의 세포독성과 멜라닌합성에 대한 감잎 추출물의 영향을 배양 B16/F10 흑색종세포를 재료로 하여 조사하였다. 또한, 감잎 추출물의 항산화능을 lactate dehydrogenase(LDH) 활성에 의하여 조사하였다. 본 연구에서 NiSO₄는 배양 B16/F10 흑색종세포에 처리한 농도에 비례하여 세포생존율을 유의하게 감소시켰으며(P<0.001), 이 때 XTT50값은 106.7μM로 중간독성인 것으로 나타났다. 또한, 항산화제인 비타민 E는 NiSO₄에 의하여 감소된 세포생존율을 유의하게 증가시킴으로서 NiSO₄의 세포독성을 방어하였다(P<0.001). 한편, NiSO₄의 세포독성에 대한 감잎 추출물의 영향에 있어서, 감잎 추출물은 NiSO4만의 처리에 비하여 유의한 세포생존율의 증가 및 LDH 활성 감소를 보임으로서 항산화능을 나타냈다. 멜라닌화에 있어서, 감잎 추출물은 NiSO₄에 의하여 증가된 티로시나제(tyrosinase) 활성과 총멜라닌양을 모두 유의하게 감소시켰다(P<0.001). 이상의 결과로부터 NiSO4의 세포독성에 산화적 손상이 관여하고 있으며, 감잎 추출물은 항산화능에 의하여 NiSO₄의 세포독성을 효과적으로 방어하였다. 또한 감잎 추출물은 티로시나제 활성과 총멜라닌양을 유의하게 감소시킴으로서 NiSO₄에 의한 멜라닌화를 효과적으로 방어하였다. 따라서, 감잎과 같은 천연추출성분은 NiSO₄처럼 산화적 손상과 관련이 있는 중금속의 독성으로부터의 미백 및 항독을 위한 천연소재로서의 개발적 가치가 크다고 생각된다.
감잎 4품종과 녹차의 화학성분을 분석하고 항산화 활성을 비교하여 전통 건강차로써 감잎차의 우수성을 알리는 기초 자료로 활용하고자 건시료의 유리당, 유리아미노산, 비타민C, 카페인 등을 분석하였다. 감잎 4품종 모두 녹차에 비해 총 유리당 함량이 높았고 유리당은 sucrose, glucose, fructose, maltose, xylose 5종의 유리당이 동정되었으며 그 중 sucrose의 함량이 가장 높았다. 유리아미노산 함량은 감잎 품종에서 총
Comparing and analysing the productivity and the main ingredients of persimmon leaves by cultivars were conducted to improve the quality and productivity of the per-simmon leaf tea. As a result of the survey, Dungsi was decidedly good in growth co-ndition of survival rate, stem diameter and the number of shoots etc. Also , DUngsi had the highest leaf productivity of 567 leaves and the leaf weight of 1.98 kg per one plant after three years of planting . On accounts of productivity, ingredient analysis and panel test etc. we collectively through that Dungsi was the best cutltivar for the persimmon leaf tea.
Changes in antioxidative substance levels in eleven different cultivars of persimmon leaves during growth were investigated. In general, the contents of soluble phenols, L-ascorbic acid and flavonoids in astringent persimmon leaves(APL) were higher than those of nonastringent persimmon leaves(NAPL). The soluble phenol contents in APL and NAPL showed a tendency to decrease throughout leaf growth. L-ascorbic acid content in APL decreased rapidly during growth, whereas its content in NAPL reached its highest value at the late of July, and then decreased rapidly. Major flavonoids in APL and NAPL were quercetin and Kaempferol which were present in conjugate forms. Before acid hydrolysis, the contents of kaempferol and quercetin in APL and NAPL remained at a relatively constant level until the late of July, and then decreased slightly. After acid hydrolysis, kaempferol contents in APL and NAPL varied significantly by cultivar and growth stage, while quercetin contents decreased slowly until the late of July, and then increased drastically, reached a maximum at the early of August, afterward continuously decreased. These results suggest that APL harvested at the early of June may be useful as potential sources of natural antioxidants.