국산 미생물 발효차의 폴리페놀 색소성분들인 데아플라빈(TF)과 데아루비긴(TR) 및 EGCG를 macrophage cell line (RAW264.7에 적용하여 nitric oxide 합성 및 사이토카인 생성을 평가하였다. 사이토카인 생성은 TF, TR 및 EGCG를 RAW264.5 cell에 적용하였을 때, $80\;{\mu}g/ml$ 농도에서 대조군과 LPS 촉진 처리에 비하여 nitric oxide 생성은 약 1.5배 증가하였다. IL-6, $TNF-{\alpha}$ 및 GM-CSF는 TF, TR 및 EGCG 농도에 의존적으로 증가하였다. $TNF-{\alpha}$ 생성은 크게 증가하였으며, 이는 TF, TR 및 EGCG가 사이토카인 생성을 통하여 면역증강 효과를 가질 것으로 나타났다 TF, TR 및 EGCG는 총 페놀 함량에 비례하여 항산화능을 나타내었으며, 암세포 증식을 유의적으로 억제하였다. 이들 폴리페놀물질의 억제효과는 그 성분들의 항암촉진작용 및 항산화활성에 의한 것으로 판단된다.
국산 미생물 발효차의 폴리페놀 색소성분들인 데아플라빈(TF)과 데아루비긴(TR) 및 EGCG를 macrophage cell line (RAW264.7에 적용하여 nitric oxide 합성 및 사이토카인 생성을 평가하였다. 사이토카인 생성은 TF, TR 및 EGCG를 RAW264.5 cell에 적용하였을 때, $80\;{\mu}g/ml$ 농도에서 대조군과 LPS 촉진 처리에 비하여 nitric oxide 생성은 약 1.5배 증가하였다. IL-6, $TNF-{\alpha}$ 및 GM-CSF는 TF, TR 및 EGCG 농도에 의존적으로 증가하였다. $TNF-{\alpha}$ 생성은 크게 증가하였으며, 이는 TF, TR 및 EGCG가 사이토카인 생성을 통하여 면역증강 효과를 가질 것으로 나타났다 TF, TR 및 EGCG는 총 페놀 함량에 비례하여 항산화능을 나타내었으며, 암세포 증식을 유의적으로 억제하였다. 이들 폴리페놀물질의 억제효과는 그 성분들의 항암촉진작용 및 항산화활성에 의한 것으로 판단된다.
Theaflavins (TF) and thearubigins (TR) are constituents of tea pigments which are polyphenols derived from Korean fermentation tea. After TF, TR and [(-) epigallocatechin-3-gallate](EGCG) have been applied to macrophage cell line (RAW264.7) nitric oxide (NO) synthesis and cytokines production were e...
Theaflavins (TF) and thearubigins (TR) are constituents of tea pigments which are polyphenols derived from Korean fermentation tea. After TF, TR and [(-) epigallocatechin-3-gallate](EGCG) have been applied to macrophage cell line (RAW264.7) nitric oxide (NO) synthesis and cytokines production were estimated. Cytokines production by enzyme linked immune-sorbent assay (ELISA) determined. NO production was increased by about 1.5-folds at the dose of $80\;{\mu}g/ml$ compared to control and lipopolysaccharide (LPS) stimulation when TF, TR and EGCG were applied to a RAW264.7 cell. Interleukin-6 (IL-6), Tumor necrosis factor ($TNF-{\alpha}$) and granulocyte-macrophage colony stimulating factor (GM-CSF) increased depended on concentrations of TF, TR and EGCG. The production of tumor necrosis $factor-{\alpha}$ increased highly in TR, TF and EGCG group with LPS. These results suggest that TF, TR and EGCG have immune-enhancement effect through the cytokine production. TF, TR and EGCG inhibited cancer cell viability, the anticancer effect of these polyphenols may explain the anti-tumor promotion action and antioxidant activity of these tea constituents.
Theaflavins (TF) and thearubigins (TR) are constituents of tea pigments which are polyphenols derived from Korean fermentation tea. After TF, TR and [(-) epigallocatechin-3-gallate](EGCG) have been applied to macrophage cell line (RAW264.7) nitric oxide (NO) synthesis and cytokines production were estimated. Cytokines production by enzyme linked immune-sorbent assay (ELISA) determined. NO production was increased by about 1.5-folds at the dose of $80\;{\mu}g/ml$ compared to control and lipopolysaccharide (LPS) stimulation when TF, TR and EGCG were applied to a RAW264.7 cell. Interleukin-6 (IL-6), Tumor necrosis factor ($TNF-{\alpha}$) and granulocyte-macrophage colony stimulating factor (GM-CSF) increased depended on concentrations of TF, TR and EGCG. The production of tumor necrosis $factor-{\alpha}$ increased highly in TR, TF and EGCG group with LPS. These results suggest that TF, TR and EGCG have immune-enhancement effect through the cytokine production. TF, TR and EGCG inhibited cancer cell viability, the anticancer effect of these polyphenols may explain the anti-tumor promotion action and antioxidant activity of these tea constituents.
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대상 데이터
TNF- a, IL-6 and GM-CSF (Pharmingen, San Diego, USA) levels in macrophage culture medium were determined by enzyme-linked immunosorbent assay. DMEM (Dulbeco's modified Eagle's medium), FBS (fetal bovine serum), 0.05% trypsin-0.02% EDTA and 100 units/ml perdcillin-strepto- mycin were purchased from GIBCO Co. (Grand Island, NY, USA). RAW264.
(Grand Island, NY, USA). RAW264.7 cells and cancer cells (HT-29, A-549, Hela and HepG2) were purchased from the Korean Cell Line Bank (Seoul, Korea). TF and TR were extracted from Korean microbial fermented tea, which was donated from the Korea Fermented Food Research Institute (KOFRI) and local tea-producing industries.
2 μm filter prior to analysis in the cell test. The test cells used in this assay were A-549, HT-29, HeLa and HepG2 cells. Cells were maintained as adherent cell cultures in DMEM medium supplemented with 10% FBS and 10 units of penicillin and 10 μg/ml streptomycin at 37 ℃ in a humidified incubator containing 5% CO2.
데이터처리
The experiments shown are a summary of the data from at least three experiments and are presented as the mean± SD. Statistical significance of the data was evaluated by the ANOVA. The value of p<0.
이론/모형
7 (5xl04 cells/well) were cultured either in medium alone or in medium that contained LPS (10 ㎍/ml) or TR, TF and EGCG for the production of nitric oxide. After 48 hr of culture, the amounts of nitrite production were measured by the Griess method as described under Materials and Methods. *p<0.
Anticancer activity was measured using the SRB assay [23]. All samples were sterilized by filtration through a 0.
Total TF were determined by the Flavognost method [11]. The percentage of dry matter in the sample was determined by oven-drying.
성능/효과
In this study, TF, TR and EGCG (10—80 μg/ml) increased NO, IL-6 and GM-CSF production, TNF-a production tend to increased highly in EGCG treatment. NO has been identified as a major effecter molecule produced by macrophages and is involved in the regulation of apoptosis and in host defenses against microorganisms and tumor cells [5].
EGCG was more effective in inhibiting the growth of these cancer cell lines than TF and TR at 80 pg/ml. The observed inhibition rate of TR, TF and EGCG were 74.2%, 76.6% and 81.1% for HeLa at 80 μg/ml, respectively, whereas those for TR, TF and EGCG were 68.1%, 69.7%, 89.7%, 74.7%, 70.7% and 82.4% for HT-29 and HepG2 at 80 pig/ml, respectively (Table 4, 5).
후속연구
Also, more detailed work is required on the protective effects against cancer cell and immuno-modulating effects to know their exact mechanism of action.
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