• 한방안이비인후피부과학회지
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• 제23권1호
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• pp.23-43
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• 2010

Objective : This study was performed to assess the effects of Aloe and Violae herba extracts on skin disease and skin beauty. Methods : Anti-oxidant effects were measured by the scavenging for DPPH radical, xanthine oxidase activity. Anti-inflammatory effects were examined by relations in NO synthesis, IL-$1{\beta}$, IL-6, TNF-$\alpha$, NF-kB, COX-2, MAP kinase. The skin wrinkle formation effects were measured by collagenase and elastase activities. The whitening effects were examined by tyrosinase activities, melanin synthesis in MNT-1 cell. Results : 1. In an anti-oxidant test, Aloe and Violae herba extracts showed high radical scavenging activity. 2. In an anti-inflammatory test, Aloe and Violae herba extracts strongly inhibited the lipopolysaccharide(LPS)-induced nitric oxide(NO) release from the RAW 246.7 macrophage cells. Aloe and Violae herba extracts also inhibited the LPS-induced IL-$1{\beta}$ and COX-2 expressions. The inhibitory effects of Aloe and Violae herba extracts on macrophage activation were via the inhibition of NF-kB, evidenced by transient transfection assay. Furthermore, Aloe and Violae herba extracts weakly inhibited the activation of Jun-N-terminal kinase(JNK) but they did not have any effects on p38 MAP kinase in RAW 264.7 cells. 3. In the skin wrinkle formation assay, Aloe extract strongly inhibited collagenase and elastase, whose activity are tightly related with the wrinkle formation. 4. In the skin whitening assay, Aloe and Viloae herba extracts weakly inhibited tyrosinase activity, however, it was not statistically significant. Besides they did not have any effects on melanin synthesis, indicating that they could not be applicable for skin whitening. Conclusion : This study show that Aloe and Violae herba extracts may play a significant role in skin disease and skin beauty.

• 대한화장품학회지
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• 제25권2호
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• pp.59-75
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• 1999

Skin is continuously exposed to external stimuli including ultraviolet radiation, which is a major cause of skin photoaging. According to recent discoveries, UVA with a lower energy but deep-penetrating properties, compared to UVB, is likely to play a major part in causing skin photoaging. The clinical and histochemical changes of photoaging are well characterized, but the biochemical mechanisms are poorly understood partly due to the lack of suitable experimental systems. In this work, three-dimensional, reconstituted skin culture models were prepared. After certain period of maturation, the equivalent models were shown to be similar in structure and biochemical characteristics to normal skin. Mature dermal and skin equivalent models were exposed to sub-lethal doses of UVA, and the effects of UVA relevant to dermal photoaging were monitored, including the production of elastin, collagen, collagenase(MMP-1), and tissue inhibitor of metalloproteinases-1 (TIMP-1). Interestingly, dermal and skin equivalents reacted differently to acute and chronic exposure to UVA. Elastin production was increased as soon as one week after commencing UVA irradiation by chronic exposure, although a single exposure failed to do so. This early response could be an important advantage of equivalent models in studying elastosis in photoaged skin. Collagenase activity was increased by acute UVA irradiation, but returned to control levels after repeated exposure. On the other hand, collagen biosynthesis, which was increased by a single exposure, decreased slightly during 5 weeks of prolonged UVA exposure. Collagenase has been thought to be responsible for collagen degeneration in dermal photoaging. However, according to the results obtained in this study, elevated collagenase activity is not likely to be responsible for the degeneration of collagen in dermal photoagig, while reduced production of collagen may be the main reason. It can be concluded that reconstituted skin culture models can serve as useful experimental tools for the study of skin photoaging. These culture models are relatively simple to construct, easy to handle, and are reproducible Moreover the changes of dermal photoaging can be observed within 1-4 weeks of exposure to ultraviolet light compared to 4 months to 2 years for human or animal studies. These models will be useful for biochemical and mechanistic studies in a large number of fields including dermatology, toxicology, and pharmacology.

• 생명과학회지
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• 제16권6호
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• pp.964-971
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• 2006

기질의 침윤과 전이를 특징으로 하는 악성종양 세포는 세포외 기질이나 기저막에 의존적으로 작용한다. 세포외 기질을 분해하는 효소인 matrix metalloproteinase (MMP) 계들의 발현 및 활성증가는 대부분의 악성종양세포에서 전이와 침윤을 촉진시킨다. MMP family 가운데 특히 type IV collagenase 활성을 지닌 MMP-2와 MMP-9은 세포외기질의 중요한 구성분인 collagen, fibronectin을 분해하는 특성을 가지며 암 전이를 용이하게 하는 주요한 효소로 잘 알려져 있다. 본 연구에서는 항암후보물질인 disulfiram이 골 육종 (U20S), 신장암 (Caki-1) 및 자궁암 (Caski) 세포에서 MMP-2와 MMP-9의 효소활성 및 발현억제에 대해 조사하였다. MTT assay를 이용하여 disulfiram에 대한 암세포 viability 실험에서는 disulfiram이 암세포의 viability를 저해하였다. 또한 zymography, western blot 및 RT-PCR 등을 이용한 type IV collagenase의 활성 및 발현 실험에서 disulfiram은 type IV collagenase의 활성을 비롯하여 단백질 및 mRNA 발현을 억제시키는 것을 확인하였다. 따라서 disulfiram이 MMP-2와 MMP-9의 활성 및 발현 억제 기전을 통하여 골 육종, 신장암 및 자궁경부암 세포의 작용을 억제한다는 연구 결과는 disulfiram이 각종 악성종양의 침윤과 전이를 억제 또는 방지하기 위한 치료물질로서 임상에서 활용할 수 있는 가능성을 보여준다.

• 생명과학회지
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• 제29권11호
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• pp.1192-1199
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• 2019

본 연구는 화장품 소재로서 우절 에탄올추출물의 가능성을 확인하기 위한 것이다. 이를 위해 우리는 연근의 마디부분을 70%에탄올로 추출하여 사용하였으며 미백효능 및 주름개선 효과에 대한 생물학적 활성 평가를 수행하였다. 시료의 미백활성을 알아보기 위해 멜라닌세포(B16F10 cells)의 MTT assay를 이용한 샘플의 독성평가와 멜라닌 생성에 영향을 주는 관련 단백질의 발현량을 확인하였다. 시료의 주름활성억제 평가를 위해 collagenase inhibition assay를 수행하였다. 또한 UVB로 유도된 섬유아세포(CCD-986sk cells)의 MTT assay를 이용한 샘플의 독성평가와 matrix metalloprotease (MMP-1) inhibition 및 procollagen synthesis를 평가하였다. 미백활성 평가를 알아보기 위한 western bolt 결과, 멜라닌 생성과 관련된 두 단백질 Tyrosinase related protein-1 (TRP-1, Tyrosinase related protein-2 TRP-2)의 합성이 농도의존적으로 감소됨을 나타내었다. 게다가 우절 에탄올추출물의 collagenase inhibiion 실험의 결과, $500{\mu}g/ml$의 농도에서 대조군인 EGCG와 효능이 비슷한 80% 이상의 효과를 나타내었다. Procollagen synthesis 결과 UVB 자극군에 의해 콜라겐 합성은 68.8%로 감소되었으며 시료의 $25{\mu}g/ml$의 농도에서 80.2%의 콜라겐 합성에 회복능을 보였다. MMP-1 inhibition 실험결과는 $25{\mu}g/ml$ 농도에서 20.2%의 저해능을 나타내었다. 실험 결과는 NRN의 미백 및 주름 억제 효과를 검증하고, 향후 안전한 천연 화장품 재료로 사용될 수 있음을 확인했다.

• 대한본초학회지
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• 제27권2호
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• pp.43-46
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• 2012

Objectives : Sanguisorbae Radix(SO) is a plant in the family Rosaceae, which grows widely in open fields Korea. It has been used as traditional medicine for thousands of years, as a treatment for anti-inflammatory and it is widely used for throat infection, tonsilitis, conjuctivitis and lymphadentis. In this study, investigated skin antiaging and anti-bacterial by using SO fractions water, acetone and butanol, chloroform. Methods : The effects of anti-microbial on SO fractions and elastase inhibition activity, collagenase inhibition activity were experimented. Results : 1. The ethyl acetate fraction showed the strongest antimicrobial activity against Staphylococcus epidermidis. 2. The elastase inhibition rate and collagenase inhibition rate of the water fraction of SO was the highest other factions. Conclusions : From the above results, it was confirmed the SO has sufficient potentiality applying itself to industry and also SO can be utilized as antimicrobial natural materials and antiaging cosmetics.

• 대한본초학회지
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• 제31권4호
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• pp.53-60
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• 2016

Objective : The purpose of this study was to investigate anti-aging and antioxidant effects of extracts of Nelumbo nucifera leaves (NN-L) using ethanol on skin .Methods : Each part of leaves(NN-L), flowers(NN-F) and stem(NN-S) was extracted with 70% ethanol. We performed radical scavenging assay(DPPH, ABTS+, Superoxide anion radical), elastase inhibition assay, collagenase inhibition assay. NN-L extracts were tested for cell viability(MTT assay), MMP-1 inhibition and MMP-1 protein expression on CCD-986sk cells (human fibroblast line).Results : Recently, many studies have reported that elastin is also involved in inhibiting or repairing wrinkle formation, although collagen is a major factor in the skin wrinkle formation. We measured its free radical scavenging activity, elastase inhibitory activity and expression of MMP-1 (matrix metalloprotease-1) in human fibroblast cells. Among the parts of Nelumbo nucifera, NN-L showed the highest antioxidant activities and in radical scavenging. DPPH, ABTS+ and Superoxide anion radical scavenging activity of NN-L at concentration of 1,000 μg/mL were 91.43%, 99.31% and 73.7% respectively. In vitro elastase and collagenase inhibition effects of NN-L at concentration of 1,000 μg/mL was 42.8% and 55.3% respectively. The ethanol extract of NN-L showed cell viability of 95.4% in 50 μg/mL concentration. In addition, The results from Western blot assay showed that NN-L decreased the expression of MMP-1 protein in a dose-dependent manner (by up to 35.0% at 50 μM).Conclusion : The findings suggest that the NN-L great potential as a cosmeceutical ingredient with antioxidant and anti-wrinkling effects.

• 동의생리병리학회지
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• 제25권3호
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• pp.425-430
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• 2011

This study evaluated the effect of extraction conditions of yam (Dioscorea aimadoimo) on antioxidant, moisturizing, collagenase activity, proliferation, and migration. Yam has been recognized as a healthy food due to its various biological activities, such as anti-obesity, anti-constipation, anti-mutagenic activities, as well as its ability to decrease blood glucose and cholesterol levels. Electron donating ability of high temperature ethanol extract of Dioscorea aimadoimo (HDA) had shown 70.6% at 400 mg/ml, and low temperature ethanol extract of Dioscorea aimadoimo (LDA) had shown 40% at 400 mg/ml. SOD-like activities of LDA and HDA were 23% and 34% at 400 mg/ml respectively. LDA significantly reduced the activity of collagenase in a dose-dependent manner, which was higher than HDA. The water contents in LDA-treated skin and HDA-treated skin were increased by 45.63% and 38.65% than the placebo cream respectively. The cellular proliferation of human dermal fibroblast neonatal (HDFn) was evaluated by MTT and cell migration assay. Compared to control, the cell proliferation was elevated to 109.7% and 114% by the treatment of LDA and HDA respectively at the concentration of 200 mg/ml. In addition, LDA and HDA were induced cell migration in HDFn. Our study suggests that LDA and HDA should be a very useful cosmetic ingredient, as anti-aging and skin moisturizer.

• 한방안이비인후피부과학회지
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• 제12권1호
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• pp.47-78
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• 1999

Hoichunyonggyuksan(HCYGS) and Yongseksan(YSS) are important prescriptions that have been used in oriental medicine for stomatitis and wound healing. The study was done to evaluate the inhibitory effects of cytotoxicity, formation of superoxide on the macrophage and neutrophil, prostaglandins($PGE_2$), interleukins($IL-1{\Beta}$), collagenase activity and synthesis of collagen and DNA. The results were obtained as fo1lows: 1. HCYGS and YSS were not showed the proliferation difference of human fibroblast and monocyte in all concentrations to be experimented and in result, it was concluded that they have no cytotoxicity. 2. YSS inhibited the formation of superoxide to $48\% at the concentration of $0.001\%$ in the mouse monocyte. 3. HCYGS inhibited the formation of superoxide to $13\%$ at the concentration of $0.001\%$ as compared with control in the human monocyte. 4. HCYGS and YSS inhibited the formation of superoxide to $25\%\;and\;35\%$ respectively at the concentration of $0.0001\%\;and\;HCYGS\;showed\;38\%$ inhihitory rate on the formation of superoxide at concentration of $0.001\%$ in the human neutrophil. 5. The concentration of inhibiting the production of prostaglandins($PGE_2$) to $11\%$ in the human monocyte stimulated with E. coli were $0.01\%$ of HCYGS. 6. The concentration of inhibiting the production of interleukins($IL-l{\Beta}$) to $43\%\;and\;39\%$ in the human monocyte stimulated with E. coli were $0.01\%$ of HCYGS and YSS. 7. HCYGS and YSS didn't influence on collagen synthesis and total protein in fibroblasts. 8. HCYGS and YSS inhibited the collagenase activity to $22\%\;and\;19\%\;at\;0.1\%,\;45\%\;and\;56\%\;at\;0.2\%,\;57\%\;and\;52\%$ at $0.5\%$ respectively, but YSS exerted the inhibitory effect on collagenase activity to $27\%$ at the lower concentration of $0.01\%$. 9. HCYGS and YSS didn't show the faster recovary than control group hut exerted the consistant effect on the anti-inflammations and the formation of granulation tissue.

• 공업화학
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• 제22권1호
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• pp.56-60
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• 2011

본 연구는 함초를 이용하여 기능성 화장품 개발을 위한 기초자료를 얻기 위해 추출용매에 따른 함초 추출물의 항산화 활성, tyrosinase 저해활성, elastase 저해활성, collagenase 저해활성을 검토 하였다. 여러 아미노산 중에서 glutamic acid이 가장 높았고, 그 다음으로는 proline, alanine, $\gamma$-amino-n-butyric acid, arginine 순이였다. 폴리페놀 함량 및 항산화 효과는 ethyl acetate extract > n-butanol extract > hexane extract > hot water extract > ethanol extract 순이였다. 특히 ethyl acetate를 추출용매로 사용 할 경우 폴리페놀 함량이 31.6 mg/mL였고, 항산화 활성은 82.6%였다. Tyrosinase활성 저해도는 ethyl acetate 추출물의 농도에 의존적으로 증가하였다. 특히 추출물 농도가 20에서 100 mg/mL로 증가 할 경우 tyrosinase활성 저해도는 15.2에서 98.2%로 증가 했다. Ethylacetate 추출물 농도가 20에서 100 ug/mL 증가 할 경우 elastase 저해활성은 2.9에서 32.5%로 증가 되었고, collagenase 저해 활성은 3.8에서 29.7%로 증가 하였다. 이상의 결과는 함초 ethylacetate추출물이 화장품 기능성 소재로서 응용 가능성을 나타내었다.

• 대한핵의학회지
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• 제30권3호
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• pp.332-337
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• 1996

기저막 성분인 type IV collagen을 분해하는 type IV collagenase의 농도와 종양의 전이사이에 상관관계가 있다는 동물 실험보고가 있다. 저자들은 유방암환자의 종양조직내 type IV collagenase의 농도와 골스캔상 골전이 소견과 비교하여 그 의의를 알아보고자 하였다. 원발성 및 전이성 유방암 환자의 종양조직에서 92kDa 및 72kDa type IV collagenase에 대한 면역조직화학염색을 각각 57명, 56명 환자에서 시행하여 각 효소 농도를 평가하고 골스캔상 골전이 소견을 관찰하고 등급을 부여하였다. 면역조직화학적으로 평가한 각 효소의 농도는 원발성 유방암과 전이성 유방암 환자 사이에 큰 차이가 있었으며, 골스캔 소견과 효소농도를 상호 비교한 결과 각 효소의 농도가 170이하일 경우에는 골스캔상 활동적인 골전이 소견을 볼 수 없었으나 효소의 농도가 200이상일 경우 골스캔 소견은 정상에서 골전이 소견까지 매우 다양하게 분포하였다. 결론적으로, 면역 조직화학적으로 측정한 92kDa 및 72kDa collagenase의 농도가 170이하일 때는 골스캔상 대부분 정상소견을 보여 골전이의 확률이 낮았다. 반면에 각 효소치의 농도가 200이상일 경우에는 골전이의 확진과 병소의 위치를 확인하고 추적검사를 위해서는 골스캔이 필요하다고 사료된다.