Earticle

The aim of this study is to evaluate the antioxidant, antimicrobial, and antitumor activities of various concentrations of partially purified substance(s) from green tea seed (Camellia sinensis L.). The total polyphenol contents of each fraction (non-adsorption fraction: F-1, fraction eluted with 40% methanol: F-2, and fraction eluted with 100% methanol: F-3) purified by Diaion HP-20 column chromatography were, in the increasing order: F-1 (3.7 mg tannic acid equivalents, TAB/g) < F-3 (23.2 mg TAB/g) < seed extracts (26.2 mg TAB/g) < F-2 (42.7 mg TAB/g). The scavenging activities toward the 1,1-diphenyl-2-picyrylhydrazyl (DPPH) radical were, in decreasing order: F-2 (93.3%) > butylated hydroxytoluene (BHT; 89.8%) > ascorbic acid (89.3%) > leaf extracts (70.3%) > F-3 (15.9%) > seed extracts (15.8%) > F-1 (14.8%) at a 0.1% concentration. In studies on antimicrobial activities, the results indicate that the growth of yeast (Candida albicans KCCM 11282 and Cryptococcus neoformans KCCM 50544) was inhibited more so than that of other fungi (Alternaria alternate KCTC 6005 and Rhizoctonia solani). In addition, it appears that the antitumor activities of the F-1, F-2, and F-3 fractions at a concentration of 50 ug/mL showed 6, 7, and 23% growth inhibition of the HEC-1B cell line, 14, 11, 82% inhibition of the HEP-2 cell line, and 8, 16, and 81% inhibition of the SK-OV-3 cell line, respectively. Overall these results indicate that the antioxidant activity is greatest in the F-2 fraction, and the antimicrobial and antitumor activities are greatest in the F-3 fraction.

We investigated the catechin content in green tea leaf (GTL) and green tea seed (GTS), the antiproliferative and detoxifying phase II enzyme-inducing activities of the methanolic (80%, v/v) extracts from GTL and GTS. GTL and GTS contained 8,685±1,061 and 108±32 ug/g epigallocatechin gallate (EGCG), 11,486±506 and 116±72 ug/g epigallocatechin (EGC),3,535±308 and 821±95 ug/g epicatechin gallate (ECG), and 1,429±177 and 37±44 ug/g epicatechin (EC), respectively. The methanolic extract of GTS showed a greater increase in quinone reductase activity and antiproliferation potential against mouse hepatoma cells than GTL extract did. GTS treatment resulted in the accumulation at sub-G1 phase of mouse hepatoma hepa1c1c7 cells as assessed by flow cytometry. Enhancement of phase II enzyme activity by GTS extract was shown to be mediated, directly or indirectly, via interaction with the antioxidant response element (ARE) sequence in the genes encoding the phase enzymes. As the catechin content in GTS was significantly lower than that in GTL, components other than catechins appear to be responsible for the anticarcinogenic activity of the seed. In summary, these results suggest that the 80% methanolic extract of GTS deserves further study to evaluate its potential as an anticarcinogenic agent and to investigate its mechanism of action.

For investigating anti-cancer activity of GSE, anti-cytotoxicity, anti-oxidative capacity and anti-angiogenic effect were experimented in vitro with HUVECs cell. Anti-angiogenic effect of 0.005% of GSE was showed 7 times(92.0%) higher than that of 25uM EGCG(12.0%), and Cytotoxicity on HUVECs was very low at 10.0x10-3% of GSE. 4 kinds of cancer cells such as, SNU-423(liver cancer cell), HCC-1428(brast cancer cell) and SNU-1005(uterine cancer) were controlled by 5.0%, 36.7% and 94.8% on stomach cancer cell, by 36.8%, 57.9% and 79.0% on liver cancer cell, by 20.0%, 40.0% and 56.7% on brast cancer, and 29.4%, 52.9% and 80.0% on uterine cancer with GSE concentration of 0.5x10-4%, 5.0x10-3%, and 10.0x10-3%, respectively.

GSE inhibited cell adhesion when both cell types were pretreated with GSE, the adhesion of U937cells to IL-1β-stimulated HUVECs was completely suppressed by 57.1%, 94.2%, 100.0% and 105.7% at different concentration of 5.0, 12.5, 25.0, 50.0 x 10-5%, respectively. For inquiring anti-angiogenesis mechanism, when western blot was performed with different dose of GSE extract, signal molecules of VEGFR-2, β-catenin and PI3-K were suppressed. As the signal transduction from VEGFR-2, β-catenin and PI3-K to NF-kB was interupted, angiogenesis could not occured causing not activated NF-kB. SNU-719(stomach cancer cell), SNU-423(liver cancer cell), HCC-1428(brast cancer cell) and SNU-1005(uterine cancer) were tranplanted into nude mouse, and then observed their change of grafts. While, the grafites treated GSE with abdominal injection for 4 week into nude mouse tranplanted 4 kinds of cancer cells were perfectly necrosed. GSE is a useful substance for developing therapeutics of angiogenesis related diseases such as cancer, metastasis, rheumathioid arthritis and obesity.

Stained and discolored hair will be damaged by the shampooing, daily UV disposal, and the use of hair dryer. Thus many studies about the effect of various natural substances on the re-secure the skin and scalp are recently reported. This study was carried out to investigate the effect of green tea (Camelloia sinensis) seed oil on colored (dyed) and decolored (bleached) hair. The beneficial effects of green tea seed oil are already well known, but little research has been done about the hair treatment and fade-resistant effect. Dyed and bleached hair was pretreated with green tea seed oil to determine the tensile strength and elongation of the hair, to analyze the hair surface using SEM, and to compare the color fade using spectrocolormeter. The results showed that the tensile strength increased with green tea seed oil pretreatment samples for virgin, dyed, and bleached hairs. Elongation showed the reverse results showing the presence of hair treatment effect. The results of the surface pre-treatment in all groups analyzed by SEM, the hair cuticle became sharper, so coating effect were identified with all samples. The value of the L*, a*, b* decreased with washed hairs damaged by UV irradiation and the values were decreased also in dyed and bleached hair. In summary, green tea seed oil prevent reinjury to the heat and UV rays for colored and decolored hairs. Cosmetic practice effects of the oil were identified in the field to be appropriate to the customer's skin and scalp that natural cosmetic oils would like to offer.

본 연구에서는 건조된 녹차 뿌리와 씨에 함유된 성분중에서 무기성분, 비타민, 총 아미노산과 유리 아미노산, 총 폴리페놀 및 그 구성 성분들의 함량을 분석하였다. 뿌리와 씨의 무기성분 중에서는 K의 함량이 각각 1,052 ㎎/100 g과 1,480 ㎎/100 g으로 가장 많았고, 뿌리에서 P, Ca, Mg, Na, Fe, Mn, Zn, Cu의 순으로, 씨에서는 P, Na, Ca, Mg, Mn, Fe, Cu, Zn의 순으로 함량이 많았다. 뿌리에서 vitamin C의 함량은 5.72 ㎎/100 g으로 가장 많았고 씨에서는 6.05 ㎎/100 g으로 나타났다. 비타민의 함량을 살펴 본 결과 녹차 뿌리보다 씨에 더욱 많은 종류의 vitamin이 함유되어 있었고 그 함량도 뿌리에 비해 다소 높은 경향이었다. 총 아미노산은 뿌리에 1,651 ㎎/100 g, 씨에 4,335 ㎎/100 g 함유되어 있어 각국 시판 녹차에 비해 그 함량이 높았고, 녹차에서 존재하지 않는 아미노산인 phenylalanine도 16.00 ㎎/100 g와 139.00 ㎎/100 g 존재하는 것으로 나타났다. 유리 아미노산의 경우, 뿌리에서 녹차의 품질을 평가하는 theanine이 전체 유리아미노산 함량의 48%를 차지하였고 쓴맛을 내는 arginine, valine 및 tryptophan 등의 함량이 높았으며, 열매에서는 단맛과 감칠맛을 내는 glutamic acid, alanine, theanine, aspartic acid 및 serine 등의 함량이 많았다. 총 polyphenol은 뿌리에 237 ㎎/100 g으로 씨에서의 함량 81 ㎎/100 g보다 3배가량 높았다. Polyphenol 중 catechin의 함량은 뿌리에서 66 ㎎/100 g, 씨에서 1 ㎎/100 g으로 나타났고 caffeic acid는 뿌리에서 0.78 ㎎/100 g으로 나타났지만 씨에서는 검출되지 않았다. 또한 chlorogenic acid는 뿌리와 씨에서 각각 3.7 ㎎/100 g과 6.6 ㎎/100 g 검출되었다.

Green tea is popular plants in Asian countries and has become more widespread in western countries due to its taste characteristics and health benefits. Apart from green tea leafs, however, the use of root and seed of green tea tree has not intensively been investigated yet. In this study, the contents of mineral, vitamin, total amino acid, free amino acid, and total polyphenol (catechin, caffeic acid, and chlorogenic acid) of the root and seed of green tea tree were analyzed for the development of healthy foods. For minerals, potassium contents were 1,052 and 1,480 mg/100g-dry weight of root and seed, respectively. The order of mineral contents were as follows: K > P > Ca > Mg > Na > Fe > Mn > Zn > Cu in root and K >P > Na > Ca > Mg > Mn > Fe > Cu > Zn in seed. For vitamins, vitamin C contents were 5.72 and 6.05 mg/100g-dry weight of root and seed, respectively. The presence of more various kinds of vitamins were observed in seed than in root. For total amino acids, the contents were 1,651 and 4,335 mg/100g-dry weight of root and seed, respectively. The total amino acid contents of seed and root were higher than those in commercial green tea products. Especially the phenylalanine contents were 16 and 139 mg/100g-dry weight of root and seed, respectively whereas phenylalanine was not found in commercial green tea products. Concerning free amino acids, the bitter tasting amino acids such as arginine, valine and tryptophan were more abundant in root and the sweet tasting ones such as glutamic acid, alanine, aspartic acid, and serine were more abundant in seed. The total polyphenol contents were 237 and 81 mg/100g-dry weight of root and seed, respectively. The polyphenol contents in root were three times higher than that in seed so root may be a better source for antioxidant ingredients than seed. Among many polyphenols, catechin, caffeic acid, and chlorogenic acid were the top three major components.

The aim of this study was to investigate the tyrosinase inhibitory activity, L-DOPA auto-oxidation inhibitory activity, melanogenesis inhibition, and cell viability after UV irradiation for whitening functional cosmetic materials using green tea seed extract. Tyrosinase and L-DOPA auto-oxidation inhibitory activities, melanogenesis inhibition, and cell viability showed a dose-dependent increase. The inhibitory activities of tyrosinase in B16 melanoma cells using 300 μg/mL of ethanol extract was similar to Arbutin as positive control. In the case of the L-DOPA auto-oxidation inhibitory activity, when using hot water extract, it showed 40.3% at 300 μg/mL. On the other hand, with the ethanol extract, it was 49.2% at 300 μg/mL. The melanogenesis inhibition was 40.3% at 300 μg/mL of hot water extract. On the other hand, with the ethanol extract, it was 49.2% at 300 μg/mL. The melanogenesis was about 43% decrease compared to positive control group with α-MSH. The human skin fibroblasts viability was 83.2% at 300 μg/mL of hot water extract. On the other hand, with the ethanol extract, it was 86.7%. These results made it possible to expect the application value as a functional cosmetic material with whitening effect that can be industrialized by extract of green tea seed using Iksan Ungpo Region.

BACKGROUND/OBJECTIVES: Ultraviolet B (UVB) irradiation on skin can induce production of reactive oxygen species (ROS), which cause expression of matrix metalloproteinases (MMPs) and collagen degradation. Thus, chronic exposure of skin to UVB irradiation leads to histological changes consistent with aging, such as wrinkling, abnormal pigmentation, and loss of elasticity. We investigated the protective effect of the standardized green tea seed extract (GSE) on UVB-induced skin photoaging in hairless mice. MATERIALS/METHODS: Skin photoaging was induced by UVB irradiation on the back of Skh-1 hairless mice three times per week and UVB irradiation was performed for 10 weeks. Mice were divided into six groups; normal control, UVB irradiated control group, positive control (UVB + dietary supplement of vitamin C 100 mg/kg), GSE 10 mg/kg (UVB + dietary supplement of GSE 10 mg/kg), GSE 100 mg/kg (UVB + dietary supplement of GSE 100 mg/kg), and GSE 200 mg/kg (UVB + dietary supplement of GSE 200 mg/kg). RESULTS: The dietary supplement GSE attenuated UVB irradiation-induced wrinkle formation and the decrease in density of dermal collagen fiber. In addition, results of the antioxidant analysis showed that GSE induced a significant increase in antioxidant enzyme activity compared with the UVB irradiation control group. Dietary supplementation with GSE 200 mg/kg resulted in a significant decrease in expression of MMP-1, MMP-3, and MMP-9 and an increase in expression of TIMP and type-1 collagen. CONCLUSIONS: Findings of this study suggest that dietary supplement GSE could be useful in attenuation of UVB irradiation-induced skin photoaging and wrinkle formation due to regulation of antioxidant defense systems and MMPs expression.

Green tea seed was extracted with absolute ethanol,and the green tea seed extract(GTSE)was subjected to assays for toxicity, antioxidant ability, angiogenesis inhibitory effects and cell adhesion, as well as western blotting, and an in vivo experiment against 4 high-ranking adult cancers in Korea. Our series of experimental data demonstrated that GTSE has an antioxidant ability superior to that of EGCG in the green tea leaf, and also exhibits a profound high tumor growth inhibitory activity on a variety of cancer cell lines, as well as nude mice infected with cancer cells. GTSE was identified as a natural anticancer compound showing excellent angiogenesis inhibition and cancer cell suppression abilities. Our preliminary observations also indicate that GTSE may be another potential source of natural dietary antioxidants and also may be applicable as a novel natural anticancer agent.

To evaluate the skin cell protective and anti-wrinkle efficacy of hot water extract (HWE) from green tea seed shells, this study investigated the extraction yield, total flavonoid and polyphenol contents, cell protection, reactive oxygen species (ROS) scavenging effect, the inhibition of 3,4-dihydroxyphenylalanine (DOPA) oxidation, induction of keratin formation, collagen synthesis, and improvement in wrinkle. The yield of the green tea seed shell HWE was 2.1 g/g, with total polyphenol and flavonoid contents of 18.26 mg/g and 2.87 mg/g, respectively. The cell viability exhibited low cytotoxicity at concentrations below 90 μg/ml and provided protective effects against H2O2-induced oxidative stress. The ROS scavenging activity increased in a dose-dependent manner, reaching 49.3% at 70 μg/ml. In the case of DOPA oxidation, it inhibited by 46.2%, suggesting a whitening effect. Moreover, the expression of keratin and collagen synthesis in keratinocytes was enhanced in a concentration-dependent manner. Specifically, the collagen synthesis level in the 70 μg/ml treatment group reached 57.6% of that in the TGF-β1 positive control group. Elastase inhibition activity also increased with concentration, showing a maximum inhibition rate of 65.6% at 70 μg/ml, suggesting potential anti-wrinkle effects. These results suggest that green tea seed shell HWE possesses antioxidant and wrinkle-improving properties with low cytotoxicity, and may serve as a promising functional cosmetic and health care ingredient.

This study investigated the effects of the addition of gardenia seed, green tea, or cactus pear (Opuntia ficus-indica) to rice batter at 10% on the lipid oxidation, pigments, antioxidants, and antioxidant activity of lotus root bugak and frying oil. Lipid oxidation was evaluated based on the conjugated dienoic acid and p-anisidine values. Lipid oxidation and tocopherol degradation were significantly reduced in the gardenia seed-added bugak and frying oil, whereas the cactus pear-added bugak and frying oil showed an increase. The addition of green tea had no significant effects on the lipid oxidation of bugak and frying oil. The in vitro antioxidant activity of lotus root bugak significantly increased with the addition of gardenia seed, green tea, or cactus pear. The results suggested that green tea and gardenia seed could improve the health and food functionality of antioxidation for lotus root bugak, respectively.

The present study evaluates antioxidant, tyrosinase inhibitory, and acetylcholinesterase (AChE) inhibitory activities of seed and its pericarp of tea plant (Camellia sinensis L.). Water and methanol extracts of tea seed and pericarp were prepared in a shaking incubator overnight at room temperature. The highest total phenolic contents, DPPH and ABTS radical scavenging activities, reducing power, ${\beta}$-carotene bleaching inhibition activity, highest tyrosinase inhibitory activity, and AChE inhibitiory activity were found in the methanol extracts of pericarp. Caffeine, gallic acid, several phenolic compounds, hydroxymethyl furfural, and fatty acid derived compounds were detected in the extract of tea seed and pericarp. The results indicate that seed and pericarp could be utilized as the potential resources for antioxidant ingredients in food industry. In addition, these compounds may protect Alzheimer's disease as they had inhibitory activity of acetylcholine esterase.

본 연구는 산업 부산물의 이용성 향상을 위하여 폐자원인 녹차씨 껍질 추출물의 미백 활성을 평가하기 위하여 수행되었다. 녹차씨 껍질로부터 유용성 성분을 얻기 위하여 에탄올 추출을 한 결과 건조중량 대비 1.24%의 추출물의 수율을 얻을 수 있었고, DPPH 라디칼 소거능 및 tyrosinase 억제능을 관찰한 결과 녹차씨 껍질 추출물의 농도가 증가할수록 활성이 유의적으로 증가하는 것으로 나타났다. 또한 이러한 미백 효과가 세포 내에서도 일어나는지에 관하여 알아보기 위하여 melanoma 세포인 B16BL6에 ${\alpha}$-MSH를 처리하여 melanin의 형성을 촉진시키는 melanogenesis 과정을 유도한 후 녹차씨 껍질 추출물을 처리한 결과 농도 의존적으로 melanin의 형성이 감소되는 것을 알 수 있었다. 이상의 결과로 녹차씨 껍질 추출물의 미백 활성을 확인할 수 있었으며, 이를 활용하여 식품 및 화장품의 기능성 소재로 이용한다면 산업 부산물인 녹차씨 껍질을 활용하는 차원에서 그 경제적 가치가 매우 클 것으로 사료된다.

The objective of this study was to evaluate the whitening effect of green tea seed shell as an industrial byproduct. Green tea seed shell extract (GTSE) was obtained by ethanol extraction, and the yield was 1.24%. 1,1-Diphenyl-2-picrylhydrazyl radical scavenging and tyrosinase inhibitory activity of GTSE increased dose-dependently. To estimate inhibition of melanin synthesis, viability was tested in B16BL6 melanoma cells. GTSE treatment induced cytotoxicity at a concentration higher than $125{\mu}g/mL$ but did not induce cytoxicity lower than $62.5{\mu}g/mL$. Thus, we fixed the optimal concentration at $62.5{\mu}g/mL$. Using this optimal concentration, melanin synthesis inhibition was measured, and GTSE treatment significantly reduced melanin synthesis induced by ${\alpha}$-melanin stimulating hormone. Therefore, the results indicate that green tea seed shell extracts may have potential melanin synthesis inhibitory activity and may be useful for development of whitening material as a natural ingredient.

본 연구는 아직 기능성 입증이 미비한 녹차나무 씨 추출물을 사용하여 자외선을 조사한 인체 피부 섬유아세포에서 항산화 효능과 MMPs 및 collagen의 변화를 살펴봄으로써 광노화 억제 효능을 평가하였다. 녹차나무 씨 추출물 농도별 라디칼 소거능을 살펴보기 위하여 ABTS와 DPPH 라디칼 소거능을 측정한 결과 녹차나무 씨 추출물 농도 의존적으로 소거능을 증가시켰다. 25 $mJ/cm^2$ 자외선을 조사한 인체 피부 섬유아세포에서 녹차나무 씨 추출물의 농도별(10 ${\mu}g/mL$, 30 ${\mu}g/mL$, 50 ${\mu}g/mL$) 처리가 미치는 영향을 살펴보았다. 항산화 효소(SOD, GPx, catalase) 활성의 변화를 측정한 결과에서 자외선 조사에 의하여 감소한 활성을 녹차나무 씨 추출물의 처리가 유의적으로 증가시켰음을 확인하였다. 또한 녹차나무 씨 추출물의 처리는 MMP-1의 합성을 감소시키고 collagen의 합성을 증가시켰으며, MMP-1, MMP-3, MMP-9의 mRNA 발현을 감소시키고 type-1 collagen의 mRNA 발현을 증가시켰다. 따라서 녹차나무 씨 추출물은 항산화 활성을 지녔으며 자외선 조사에 의한 활성산소종으로부터 보호하여 MMPs 발현을 감소시키고 collagen 분해를 억제시켜 피부 노화억제에 긍정적인 영향을 미쳤으며, 이러한 기능성 입증으로 녹차나무의 활용 증가를 기대할 수 있다.

In this study, we investigated the protective effects of green tea seed extract (GSE) against UVB-induced skin damage in human skin fibroblasts. GSE was first analyzed for antioxidant activity using 1,1-diphenyl-2picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radical scavenging assays. Treatment of UV-irradiated fibroblast with GSE at 10~50 ${\mu}g/mL$ significantly increased DPPH and ABTS radical scavenging activities in a dose-dependent manner. GSE treatment inhibited matrix metalloproteinase (MMP-1, MMP-3, and MMP-9) expression and MMP-1 secretion caused by UVB irradiation. Moreover, treatment with GSE significantly increased type-1 collagen expression and production. We next examined levels of antioxidative enzymes (SOD, catalase, and GPx). Reduced antioxidative enzyme activities caused by UVB irradiation were recovered by treatment with GSE at 30 ${\mu}g/mL$ and 50 ${\mu}g/mL$. In conclusion, these results show that GSE has protective effects against UVB-induced skin damage in human skin fibroblasts by regulating antioxidative defense systems and MMP expression.

본 연구는 산업 부산물의 이용성 증진을 위하여 폐자원인 녹차씨껍질 추출물의 항산화 활성 및 신경세포 보호 효과에 관하여 평가하였다. 녹차씨껍질로부터 유용성 성분을 얻기 위하여 에탄올 추출을 한 결과 약 1.44%의 추출물의 수율을 얻을 수 있었고, 항산화 활성에 관하여 평가하기 위하여 라디칼 소거능 및 xanthine oxidase 저해능, 환원력을 평가한 결과, 녹차씨껍질 추출물의 농도가 증가할수록 항산화 활성이 유의적으로 증가하는 것으로 나타났다. 또한, 이러한 녹차씨껍질 추출물의 뇌신경세포 보호 효과에 관하여 알아보기 위하여 생쥐의 해마 유래 뇌신경세포에 녹차씨껍질 추출물을 처리한 후 $H_2O_2$로 산화적인 스트레스를 유도하여 세포독성에 관하여 알아본 결과, 녹차씨껍질 추출물의 처리는 농도 의존적으로 뇌신경 세포의 생존율을 증가시켰으며, 이에 따라 항산화 효소인 SOD 활성이 증가하고 지질과산화 생성물인 MDA level이 감소한 것을 알 수 있었다. 이상의 결과들로 녹차씨껍질 추출물의 항산화 활성 및 뇌신경세포 보호 효과에 관하여 확인할 수 있었으며, 추후 어떤 메커니즘으로 신경세포를 보호하는지 추가적인 연구가 필요할 것으로 생각한다. 또한, 이러한 결과를 활용하여 녹차씨껍질을 기능성 소재로 활용한다면 폐자원인 녹차씨껍질을 재활용하는 차원에서 그 경제적 가치가 매우 클 것으로 생각한다.

The objective of this study was to evaluate the antioxidant activity of green tea seed shell as an industrial byproduct. Green tea seed shell extract (GTSSE) was obtained by ethanol extraction, and the yield was $1.4{\pm}0.22%$. The radical scavenging activities [1,1-diphenyl-picrylhydrazyl and 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid)], xanthine oxidase inhibition activity, and reducing power of GTSSE dose-dependently increased. To estimate the neuroprotective effect of GTSSE, viability was tested in HT22 mouse hippocampal cells. GTSSE treatment induced cytotoxicity at a concentration higher than $100{\mu}g/mL$ but not at a concentration lower than $50{\mu}g/mL$. Using this optimal concentration range, GTSSE treatment significantly increased cell viability in $H_2O_2$-treated HT22 cells. Further, GTSSE treatment increased superoxide dismutase activity and decreased the malonaldehyde level, a product of lipid peroxidation, in HT22 cells. Therefore, these results indicate that green tea seed shell extract may be useful for the development of antioxidant materials and have potential activity to prevent and treat neuro-degenerative diseases such as Alzheimer's disease.

The study was carried out to examine the anti-obesity effects of 40% ethanol extract from green tea seed (GS) and quantitative determination of caffeine as its major compound. The specificity was satisfied with retention time and UV spectrum by analysis of caffeine using HPLC and comparison with standard compound. It showed a high linearity in the calibration curve with a coefficient of correlation (R2) of 0.9974. The amount of caffeine in GS was about 4.649 mg/g (0.465%) in the three times analysis, and relative standard deviation (RSD) was less than 0.452% by the validated method. The anti-obesity effects of GS were evaluated by using Oil Red O staining in 3T3-L1 adipocytes and body weight, visceral fat and lipid profiles in high fat diet (HFD)-induced C57BL/6 obese mice. Our results indicated that treatment with GS dose-dependently decreased lipid accumulation contents (p<0.001). Moreover, after oral administration for 30 days feeding with HFD-induced obses mice, GS (100 and 300 mg/kg/day) produced a significant decrease in serum total cholesterol (TC), glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT) and visceral fat. Thus, the result of this study indicate that the GS may be a useful resource for the management of obesity.

녹차종자를 기능성 식품소재로서 이용하기 위하여 녹차 종자메탄올추출물의 생리활성을 녹차 메탄올추출물의 생리활성과 비교하였다. 녹차 메탄올추출물기 수소공여능은 $100{\mu}g/mL$ 농도 이상에서 50% 이상의 활성을 나타내었으나, 녹차종자 메탄올추출물은 $1000{\mu}g/mL$ 농도에서 21.86%의 활성을 나타내었다. 또한 녹차종자 및 녹차 메탄올추출물이 $1000{\mu}g/mL$ 농도로 처리된 흰쥐의 간 균질물에서 MDA의 생성량이 대조구의 86 Mol/g에 비하여 각각 60 및 50 Mol/g로 낮게 나타내어 이들 추출물은 항산화효과가 있음을 알 수 있었다. 녹차종자 및 녹차 메탄올추출물은 $1000{\mu}g/mL$ 농도에서 A549 및 SW480 세포에 대하여 각각 20%, 50% 이상의 성장 억제율을 나타내었다. 또한 녹차종자 메탄을 추출물이 $500{\mu}g/mL$의 농도 처리된 암세포는 대조구에 비하여 상대적인 세포 수의 감소되었으며, 심한 형태학적인 변화를 보여주었다. 대식세포 RAW264.7에 녹차 및 녹차종자 메탄을 추출물이 1, 10, 100및 $1000{\mu}g/mL$의 농도로 처리 하였을 때, 이들 추출물은 $100 {\mu}g/mL$ 농도까지 농도 의존적으로 NO(nitric oxide)의 생성을 유도하였으며, $100{\mu}g/mL$의 농도에서 그 농도는 각각 0.77와 2.04 uM로서 녹차종자 메탄을 추출물이 녹차 메탄을 추출물보다 그 효과가 크게 나타났다.

This study was to investigate the biological activities of green tea seed methanol extract (GTSME) and compared those of green tea methanol extract (GTME) for using green tea seed as the functional food material. The hydrogen-donating activity of GTSME was over 50% at the $100 {\mu}g/mL$ concentration the activity of GTME was 21.86% at the $1000{\mu}g/mL$ concentration compared with that of control. The MDA (malondialdehyde) production was 60 Mol/g and 50 Mol/g in the mouse liver homogenate teated with GTME and GTSME of $1000{\mu}g/mL$ concentrations, respectively, and the values were lower than 86 Mol/g of control. GTME and GTSME of $1000{\mu}g/mL$ concentration inhibited the proliferation of over 50% and over 20% in A549 and SW480 human cancer cells, respectively. The morphology transformation was shown in the cancer cells treated with GTSME of $500{\mu}g/mL$ with the decrease of cell numbers lower than that of control cells numbers. The NO production was increased in a dose dependent manner in the RAW264.7 macrophage cells treated with GTME and GTSME of 1, 10, 100 and $1000{\mu}g/mL$ concentrations, and the NO production by GTSME was $2.04{\mu}M$ at $100{\mu}g/mL$ concentration, and the value was higher than $0.77{\mu}M$ by GTME.

본 연구는 녹차씨 압착박의 활용성을 증가해 보고자 녹차씨를 압착한 후 남은 박을 추출하여 총 페놀, 조사포닌, 카테킨, 나린게닌 함량을 확인하고 3T3-L1 세포에서의 지방세포분화 억제 관련 효과를 확인하였다. 또한 기존에 체지방 감소 효능이 있는 것으로 알려진 녹차와도 비교하였다. 녹차씨 압착박 추출물과 녹차 추출물의 페놀, 조사포닌, 카테킨, 나린게닌 함량을 비교한 결과 녹차씨 압착박 추출물에서 나린게닌 함량이 높음을 알 수 있었다. 3T3-L1 세포에서 Oil Red O 염색법을 이용한 지방 축적저해 정도 및 세포내 중성지방 함량을 측정한 결과, 녹차씨 압착박 추출물에서 지방 축적저해 및 중성지방의 함량이 감소하는 것을 확인하였다. 녹차씨 압착박이 지방합성 및 에너지 조절 효소에 미치는 영향을 알아보기 위해 단백질 발현 실험을 실시한 결과, 녹차 또는 녹차씨 압착박 추출물에 의해서 AMPK 및 ACC의 인산화 증가 및 FAS의 발현이 감소하는 것을 확인 할 수 있었으며, 이러한 효능은 녹차 추출물보다 녹차씨 압착박 추출물에서 더 강하게 나타나는 것을 알 수 있었다. 이상의 결과로부터 녹차씨 압착박 추출물이 녹차 추출물 보다 지방세포분화 억제 효과가 높게 나타나는 것이 확인되어져, 녹차씨는 추후 체지방 감소 효능을 갖는 식품 소재로도 활용이 가능 할 것으로 생각되어진다.

The effect of the hot water extract of defatted green tea seed cake (GTSE) on lipid metabolism and the underlying mechanisms of lipolysis in mature 3T3-L1 adipocytes were investigated. In this study, we found that the naringenin content of GTSE was 5.5 mg/g; however, catechins were not detected. The intracellular lipid droplets were stained with Oil Red O dye and quantified. Compared to the control, lipid accumulation was significantly decreased by 52%, and intracellular triglyceride (TG) level was reduced by 33% after treatment with GTSE at a concentration of $40{\mu}g/mL$. To determine the mechanism of reduction in TG content, we determined the level of fatty acid synthase (FAS), phosphorylation of adenosine monophosphate-activated protein kinase (AMPK), and acetyl-coenzyme A carboxylase (ACC) in the cell model. Incubation of the 3T3-L1 adipocytes with GTSE stimulated AMPK and ACC phosphorylation in a dose-dependent manner, and decreased the expression of FAS.