• Journal of Mushroom
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• v.16 no.4
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• pp.324-330
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• 2018

The objective of this study was to evaluate antioxidant effect and tyrosinase inhibitory activity of white beech mushroom (Hypsizygus marmoreus) extracts. The white beech mushroom was extracted into hot water and methanol. Total polyphenol content was highest in the hot water extract ($8.4{\pm}3.27mg\;GAE/g$) compared to the methanol extract ($7.3{\pm}2.85mg\;GAE/g$). The flavonoids contents in hot water and methanol extracts were $3.8{\pm}3.81ug/mg$ and $2.5{\pm}1.95ug/mg$, respectively. The tyrosinase inhibitory activity of extract was increased in a dose dependent manner and tyrosinase inhibitory activity of extract (hot water extract, 69.72%; methanol extract, 52.67% at 40 mg/ml) was lower than those of positive control 2% arbutin (96%). The DPPH radical scavenging activity of the hot water and methanol extract was 80% and 74%, respectively. Hot water extract ($63.34{\pm}1.00uM\;TE/g$) were more effective in ORAC (oxygen radical absorbance capacity) value than methanol extract ($46.33{\pm}0.48uM\;TE/g$). The toxicity of hot water and methanol extracts was investigated using WST-1 (4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulphonate) assay on the B16BL6 melanoma cells.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.16
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• pp.7053-7060
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• 2015

In the present work, novel orange peel was extracted with 100%EtOH (ethanol) and fractionated into four fractions namely F1, F2, F3, F4 which were eluted from paper chromatographs using 100%EtOH, 80%EtOH, 50%EtOH and pure water respectively. The crude extract and its four fractions were evaluated for their total polyphenol content (TPC), total flavonoid content (TFC) and radical scavenging activity using DPPH (1,1-diphenyl-2-picrylhydrazyl) assay. Their cytotoxic activity using WST assay and DNA damage by agarose gel electrophoresis were also evaluated in a human leukemia HL-60 cell line. The findings revealed that F4 had the highest TPC followed by crude extract, F2, F3 and F1. However, the crude extract had the highest TFC followed by F4, F3, F2, and F1. Depending on the values of $EC_{50}$ and trolox equivalent antioxidant capacity, F4 possessed the strongest antioxidant activity while F1 and F2 displayed weak antioxidant activity. Further, incubation HL-60 cells with extract/fractions for 24h caused an inhibition of cell viability in a concentration-dependent manner. F3 and F4 exhibited a high antiproliferative activity with a narrow range of $IC_{50}$ values ($45.9-48.9{\mu}g/ml$). Crude extract exhibited the weakest antiproliferative activity with an $IC_{50}$ value of $314.89{\mu}g/ml$. Analysis of DNA fragmentation displayed DNA degradation in the form of a smear-type pattern upon agarose gel after incubation of HL-60 cells with F3 and F4 for 6 h. Overall, F3 and F4 appear to be good sources of phytochemicals with antioxidant and potential anticancer activities.

• Journal of Korean society of Dental Hygiene
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• v.14 no.2
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• pp.287-292
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• 2014

Objectives : The purpose of the study is to examine the cell growth effect and autophagy effect of Scutellariae Radix by ethanol extract in SCC 25 cells. Methods : Cell growth inhibitory effect and autophagy induced by Scutellariae Radix were confirmed by WST-1 assay, monodansylcadaverine(MDC) stain, and flow cytometry by acridine orange(AO) stain. Results : The Scutellariae Radix treatment decreased the cell proliferation in a dose and time dependent manner. Scutellariae Radix has anticancer effects that autophagic vacuoles were apparent by MDC and AO staining in SCC 25 cells. Conclusions : Scutellariae Radix showed anticancer activity against SCC 25 cells via autophagy. The data provided the possibility that Scutellariae Radix may potentially contribute to oral cancer treatment.

• Journal of Marine Life Science
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• v.6 no.2
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• pp.58-65
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• 2021

As aging progresses, reactive oxygen species (ROS) reduces skin moisturization and collapses skin barrier function. In this study, we evaluated the efficacy of skin moisturizing and skin barrier function enhancement by extracts from halophytes using HaCaT cells. Spartina anglica (S. anglica; SAE) and Calystegia soldanella (C. soldanella; CSE), a kind of halophytes, were collected from Dongmak beach in Incheon, and extracted with 70% ethanol. At the first, we evaluated the cytotoxicity of extracts in HaCaT cell using WST-8 Kit. As a result, the other experiment was conducted by setting the concentration at which the cell viability was 90% or more. SAE and CSE showed high radical scavenging activity through ABTS assay. Expression levels of genes related to skin moisturizing and skin barrier functions, were analyzed by real-time qPCR. As a result, it showed that the expression of aquaporin 3, hyaluronan synthase 2, and transglutaminase 1 was increased by SAE treatment but not changed by CSE. Activation of extracellular signal regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 mitogen activated protein kinase was induced by SAE. These results suggest that SAE can be used as functional materials for cosmetics for skin moisturizing and barrier function enhancement.

• Journal of Mushroom
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• v.19 no.4
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• pp.322-328
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• 2021

This study was carried out to evaluate potential of Hypsizygus marmoreus (brown cultivar) as a functional food and drug materials. H. marmoreus were divided into pileus and stipe and extracted in hot water and 80% ethanol. The total polyphenol content was highest in the hot water extracts (pileus 17.15±0.19 mg of GAE g, stipe 7.37±0.16 mg of GAE/g) and pileus compared to the ethanol extracts (pileus 10.23±0.14 mg of GAE/g, stipe 3.76.±0.19 mg of GAE/g) and stipe. Also, hot water extracts of pileus from H. marmoreus (brown cultivar) was more effective DPPH, ABTS, ORAC value, reducing power than ethanol extracts and stipe extracts. The pileus and stipe extracts were confirmed to be non-cytotoxic in the mouse macrophage cell line RAW 264.7 determined by WST-1 (4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulphonate) assay. Overall, extracts of H. marmoreus (brown cultivar) was higher antioxidant activity than other mushrooms, and no cytotoxicity. Therefore, H. marmoreus (brown cultivar) showed potential as a functional food and drug materials. The brown cultivar of H. marmoreus have higher antioxidant activity than white cultivar, H. marmoreus seem to have different antioxidant activity depending on the cultivar.

• Journal of Acupuncture Research
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• v.28 no.3
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• pp.101-110
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• 2011

목적 : 이 연구는 봉독이 NF-${\kappa}$B의 활성억제를 통하여 전립선 암세포주인 DU-145 세포의 성장을 억제하 는지를 확인하고 그 기전을 살펴보고자 하였다. 방법 : 봉독을 처리한 후 DU-145의 성장억제를 관찰하기 위해 WST-1 assay를 시행하였고, 세포자멸사의 관찰에는 DAPI staining assay를 통한 세포형태관찰을 시행하였으며, 염증관련유전자 발현 관찰에는 western blot analysis를 시행하였고, 세포자멸사와 연관된 NF-${\kappa}$B의 활성 변화를 관찰하기 위해 EMSA와 luciferase assay를 시행하였으며, DU-145에서 봉독과 NF-${\kappa}$B의 상호작용을 관찰하기 위해 transient transfection assay를 시행하여 세포생존율과 NF-${\kappa}$B의 활성 변동을 측정하였다. 결과 : DU-145 세포에서 봉독을 처리한 후 세포성장이 억제되었으며, 염증관련유전자 발현 및 NF-${\kappa}$B의 활성의 유의한 감소를 나타내었다. DU-145 세포에서 NF-${\kappa}$B의 p50와 IKK들을 치환하여 작용기를 없애고 봉독을 처리하였을 경우에도 세포활성 및 NF-${\kappa}$B의 활성의 유의한 감소를 나타내었다. 결론 : 이상의 결과는 봉독이 NF-${\kappa}$B의 활성 억제를 통하여 인간 전립선암세포주인 DU-145의 세포자멸사를 유발함으로써 증식억제 효과가 있음을 입증한 것으로 전립선암의 예방과 치료에 대한 효과적인 치료제 개발에 도움이 될 것으로 기대된다. 다만 그 기전에서 봉독은 기존연구와 같은 NF-${\kappa}$B p50 및 IKK들의 작용기와 상호작용 이외에 다른 기전이 관여되는 것으로 심화 연구를 요한다.

• Journal of Acupuncture Research
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• v.27 no.3
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• pp.1-13
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• 2010

목적 : 이 연구는 봉약침의 봉독과 그 주요성분인 멜리틴이 NF-${\kappa}B$의 활성억제와 세포자멸사 관련 단백질의 발현 조절을 통하여 세포자멸사를 유도함으로써 전립선 암세포주인 PC-3 세포의 성장을 억제하는지를 확인하고 해당 기전을 살펴보고자 하였다. 방법 : 봉독이나 멜리틴을 처리한 후 PC-3의 성장억제를 관찰하기 위해 WST-1 assay, CCK-8 assay를 시행하였고, 세포자멸사 조절단백질의 변동 관찰에는 western blot analysis를 시행하였고, 세포자멸사와 연관된 NF-${\kappa}B$의 활성 변화를 관찰하기 위해 EMSA를 시행하였으며, PC-3에서 봉독이나 멜리틴과 NF-${\kappa}B$의 상호작용을 관찰하기 위해 transient transfection assay를 시행하여 세포생존율과 NF-${\kappa}B$의 활성 변동을 측정하였다. 결과 : PC-3 세포에 봉독이나 멜리틴을 처리한 후, 전립선암세포의 성장, 세포자멸사의 유발, 세포자멸사 관련 단백질의 발현, NF-${\kappa}B$의 활성, NF-${\kappa}B$의 p50, $IKK{\alpha}$, $IKK{\beta}$ 치환 후 NF-${\kappa}B$의 활성과 PC-3 세포 증식에 미치는 영향을 관찰하여 다음과 같은 결과를 얻었다. 1. PC-3 세포에서 봉독이나 멜리틴을 처리한 후 세포자멸사가 유도되어 세포성장이 억제되었고, 세포자멸사 관련 단백질 중 분리된 PARP, caspase-3, -9는 유의한 증가를, Bcl-2, XIAP, cXIAP2는 유의한 감소를 나타내었다. 2. PC-3 세포에서 봉독이나 멜리틴을 처리한 후 NF-${\kappa}B$의 활성은 유의한 감소를 나타내었다. 3. PC-3 세포에서 NF-${\kappa}B$의 p50, $IKK{\alpha}$, $IKK{\beta}$를 치환하여 작용기를 없애고 봉독이나 멜리틴을 처리하였을 경우에도 NF-${\kappa}B$의 활성이 유의한 감소를 나타내었다. 결론 : 이상의 결과는 봉독이나 멜리틴이 NF-${\kappa}B$의 활성 억제를 통하여 인간 전립선암세포주인 PC-3의 세포자멸사를 유발함으로써 증식억제 효과가 있음을 입증한 것으로, 전립선암의 예방과 치료에 대한 효과적인 치료제 개발에 도움이 될 것으로 기대된다.

• Journal of Acupuncture Research
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• v.24 no.2
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• pp.1-14
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• 2007

목적 : 이 연구는 Vipera lebetina turanica의 사독약침파(蛇毒藥鍼波)(Snake venom toxin, SVT)이 in vitro에서 $NF-{\kappa}B$의 활성억제와 apoptosis 관련 단백질의 발현 조절을 통하여 세포자멸사(Apoptosis)를 유도하는지 in vivo에서 또한 전립선 암세포주인 PC-3 세포의 성장을 억제하는지 살펴보고자 하였다. 방법 : SVT를 처리한 후 PC-3의 성장억제를 관찰하기 위해 WST-1 assay, CCK-8 assay를 시행하였고，Apoptosis evaluation에는 DAPI, TUNEL staining assay를 시행하였으며，Apoptosis regulatory proteins의 변화 관찰에는 western blot analysis를 시행하였고，apoptosis와 연관된 $NF-{\kappa}B$의 활성 변화를 관찰하기 위해 EMSA시행하였으며，SVT의 핵내이동을 관찰하기 위해 Immunofluorescence Staining, Confocal immunocytochemistry를 시행하였으며，전립암세포의 종양형성에는 흉선을 제거한 쥐에 Tumorigenecity study를 시행하였다. 결과 : PC-3 세포에 SVT를 처리한 후，전립선 암세포의 성장，Apoptosis의 유발，Apoptosis관련 단백질의 발현，$NF-{\kappa}B$의 활성，SVT의 PC-3세포 핵내 이동여부 및 흉선제거 후 PC-3 세포를 이식한 쥐의 종양형성과정에 미치는 영향을 관찰하여 다음과 같은 결과를 얻었다. 1. PC-3 세포에서 SVT를 처리한 후 세포성장이 억제되고，세포자멸사가 유도되며，조절인자인 p53, caspase-3, -9는 증가되었고，Bcl-2는 감소되었다. 2. PC-3 세포에서 SVT를 처리한 후 $NF-{\kappa}B$의 활성이 유의하게 감소되었다. 3. DAPI로 염색된 상태에서 SVT가 PC-3 세포의 핵내로 이통되는 것이 관찰되었다. 4. 흉선제거 후 전립선 암세포주를 이식한 쥐에서 SVT를 피내로 주입한 결과 전립선암의 크기와 무게가 유의하게 감소하였다. 결론 : 이상의 결과는 SVT가 $NF-{\kappa}B$의 활성 억제를 통하여 인간 전립선암세포주인 PC-3의 세포자멸사를 유발함으로써 증식억제 효과가 있음을 입증한 것이며，이를 재확인한 생체 연구에서의 긍정적인 결과는 향후 SVT의 전립선암의 예방과 치료에 대한 효과적인 치료제 개발에 초석이 될 것으로 기대된다.

• Journal of Life Science
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• v.26 no.8
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• pp.928-935
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• 2016

Hypsizygus marmoreus (white cultivar), also called white beech mushrooms, are edible mushrooms commercially cultivated in Korea and Japan. This study was carried out to evaluate the antioxidant properties of H. marmoreus. H. marmoreus fruit bodies were divided into pileus and stipe. The pileus and stipe were extracted into water and 80% ethanol and their antioxidant activities were analyzed. The total polyphenol content was highest in the water extract (pileus 1137.39±0.38 mg of GAE (gallic acid equivalents)/100 g, stipe 700.86±0.06 mg of GAE/100 g) compared to the ethanol extract (pileus 923.47±0.18 mg of GAE/ 100 g, stipe 324.05±0.03 mg of GAE/100 g). Ethanol extracts from pileus showed better scavenging ability on DPPH (47.32±0.23% at 10 mg/ml) and ABTS (57.33±0.10% at 10 mg/ml) than the stipe and water extract groups. Water extract from pileus were more effective in reducing power and ORAC (oxygen radical absorbance capacity) value than stipe and ethanol extract. The toxicity of water and ethanol extracts was investigated using WST-1 (Water Soluble Tetrazolium salt) assay on the mouse macrophage cell line RAW 264.7. Overall, total polyphenol content and antioxidant activities of extracts from H. marmoreus increased in a dose dependent manner while pileus was showed better total polyphenol content and antioxidant activities than stipe.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.22 no.2
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• pp.326-335
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• 2021

This study was undertaken to determine the effect of fermented soybean extract and its fractions on skin cell proliferation and growth. The extract was procured by the pepsin and Lactobacillus rhamnosus fermentation of soybean. LC-MS analysis was performed subsequent to soybean fermentation, and cell viability was measured by the WST-1 assay. Cell proliferation was observed to increase after exposing cells to the fermented soybean extract and its fractions at all concentrations tested (0~2,000 ㎍/mL). In particular, compared to the normal control group and 120 % proliferation of the EGF (epidermal growth factor) positive control group, 160~180 % cell proliferation was achieved at 800 ㎍/ml, indicating the excellent potential as an application material for skin aging inhibition and skin cell regeneration. In addition, we also examined the effects of fermented soybean extract and its fractions on wound healing ability, in HaCaT cells and fibroblasts. Our results indicate excellent cell migration abilities after treatment with fermented soybean extract and its fractions, as compared to the control treatment. Similar cell migration abilities were observed in the positive control group (EGF). Taken together, our results indicate that fermented soybean extract and its fractions (F4 and F5) exert amelioratory effects as a natural material for skin.