• 한국응용과학기술학회지
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• 제38권3호
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• pp.870-882
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• 2021

황련은 한의학에서 진정, 소염, 항균 및 해열 등의 여러 질병의 치료에 이용되어져 왔다. 본 연구에서는 UVB와 TNF-α/IFN-γ로 유도된 HaCaT 세포에서 피부장벽과 염증관련 인자에 대한 황련 열수 추출물(CCW)의 효과를 조사하고, 보습 및 항염증 소재로서의 가능성을 알아보고자 하였다. 각 질형성세포인 HaCaT 세포에서 CCW의 보습 및 항염 효과를 확인하기 위해 MTT assay를 통한 시료의 독성 평가를 실시하였다. 이를 바탕으로, HA(Hyaluronic acid) 생성량과 filaggrin의 protein 및 mRNA 발현을 측정하였다. TNF-α/IFN-γ로 자극된 HaCaT 세포에서 CCW는 농도 의존적으로 HA의 생성량을 증가 시켰고, filaggrin의 protein 및 mRNA 발현 측정 결과 CCW의 농도 증가에 따라 발현율도 증가하였다. UVB로 유도된 HaCaT 세포에서 CCW는 ROS의 생성을 감소 시켰고, 양성대조군(positive control)인 EGCG( (-)-epigallocatechin-3-gallate)와 유의한 결과를 보여 주었다. 또한, CCW는 염증성 사이토카인 TNF-α, IL-1β, IL-6, IL-8의 발현을 억제 시켰고, TNF-α/IFN-γ로 자극된 HaCaT 세포에서 피부 염증에 따른 주요 인자인 COX-2의 protein 및 mRNA 발현이 농도 의존적으로 감소되는 것을 확인할 수 있었다. 이러한 결과는, 황련 열수 추출물의 보습 및 항염증 효과를 가지는 화장품 소재로서의 가치를 제안할 수 있다.

• 한국응용과학기술학회지
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• 제38권6호
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• pp.1383-1392
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• 2021

호장근, 적하수오, 대황, 알로에 등과 같은 생약재의 주요 약리 활성 성분인 emodin은 항산화, 항균, 항염, 항암, 간보호 등에 효능이 있는 것으로 보고되었다. 본 연구에서는 emodin의 피부 질환 및 기능성 소재로서의 활용 가능성을 알아보기 위해 염증 개선과 피부장벽기능 개선 관련 활성을 확인하였다. human keratinocyte인 HaCaT 세포에 대하여 항염효과를 관찰하기 위해 cytokine억제능은 ELISA kit로, 단백질 발현은 western blot으로 확인하였다. TNF-α (10 ng/mL)/IFN-γ (10 ng/mL)로 활성화된 HaCaT 세포에서 emodin을 농도별(5, 10, 20, 40) µM로 처리한 결과 TNF-α, IL-1β 및 IL-6의 생성량은 emodin의 농도가 증가함에 따라 감소됨을 확인하였다. 염증관련 단백질인 iNOS, COX-2 발현량에 대한 실험결과에서도 emodin 20 µM 농도에서 control 대비 iNOS는 48%, COX-2는 29%가 저해됨을 확인하였다. 피부장벽기능 개선의 지표로써 filaggrin, involucrin, loricirn의 mRNA 발현 정도와 filaggrin, involucrin, loricirn의 생성량을 확인한 결과 에모딘 농도에 의존적으로 증가하는 우수한 결과가 얻어졌다. 특히 20 µM 농도에서 대조군 대비 2배의 생성량 증가를 보인 filaggrin은 천연보습인자인 NMF의 형성에 관계되는 단백질로 각질층의 보습에 중요한 역할을 하는 것으로 알려져 있다. 결론적으로 emodin의 피부 질환 및 기능성 소재로서의 활용 가능성 중 염증 개선 및 피부장벽기능 개선 소재로 활용될 수 있음을 확인하였으며 향후 추가적인 연구가 수행되면 그 활용 범위가 더 넓어질 수 있을 것으로 사료된다.

• 대한한방소아과학회지
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• 제31권3호
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• pp.14-23
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• 2017

Objectives This study is conducted to evaluate skin fat barrier formation of Hataedock using the Coptis japonica & Glycyrrhiza uralensis extract. Methods The 3-week-old NC/Nga mice were divided into 3 groups: control group (Ctrl), Hataedock-treated group that uses the Coptis japonica & Glycyrrhiza uralensis (CGT) extract, and Hataedock-treated group that uses Bifidobacterium (BBT). After 2 weeks, changes in immunohistochemicals, and skin-lipid-barrier regulators were observed for the effects of Hataedock. Results In CGT group, loricrin-positive reaction has been increased by 231%, along with involucrin-positive reaction by 90%, filaggrin-positive reaction by 143%, and ASM-positive reaction by 341% in the stratum corneum. Conclusions Hataedock, using the extract of Coptis japonica & Glycyrrhiza uralensis, increased the expression of proteins promoting keratinocyte differentiation. This leads into conclusion that Hataedock may increase the keratinocyte formation and function which promotes skin barrier formation.

• 한방안이비인후피부과학회지
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• 제25권1호
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• pp.33-54
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• 2012

Objective : This study was carried out to compare Saengmaeksan-gamibang(SG) to Saengmaeksan(SM) on the efficacy of moisturization, skin whitening, anti-inflammation, and anti-allergy in atopic dematitis using NC/Nga mice model. Methods : We assessed the effects of SG and SM on tyrosinase, filaggrin, serine-palmitoyl transferase(SPT), COX-2, AP-1 in vitro, on IgE, IL-4, IL-5, IL-6, IgM, IgG1, IFN-${\gamma}$ in vivo with NC/Nga mice. Results : 1. SG increased the tyrosinase inhibition activity and the levels of filaggrin and SPT, decreasing the level of COX-2. 2. On the other hand, SM decreased the tyrosinase inhibition activity and the levels of filaggrin and SPT, increasing the level of COX-2. 3. Serum IgE, IL-4, 5, 6, IgM, and IgG1 were decreased in both SG group and SM group compared to the control group. The decrease degree in these factors was higher in SG than in SM. 4. Serum IFN-${\gamma}$ was increased in both SG group and SM group compared to the control group. The increase degree in IFN-${\gamma}$ was higher in SG than in SM. Conclusion : As the result of the above experiments, it was proved that SG has the moisturization, skin whitening, anti-inflammation, and anti-allergy effects, which are greater than those of SM, and provides the significant efficacy on atopic dermatitis treatment.

• 한방안이비인후피부과학회지
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• 제31권2호
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• pp.11-23
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• 2018

목적 : 본 연구에서는 $TNF-{\alpha}$와 $IFN-{\gamma}$로 자극한 인간피부각질형성세포 (HaCaT keratinocytes) 모델을 사용하여 지모가 피부장벽 기능에 미치는 영향을 알아보고자 하였다. 방법 : MTT assay를 통하여 지모 주정(70% 에탄올) 추출물 (EAA)이 HaCaT keratinocytes의 세포생존율에 미치는 영향을 확인하였으며 wound healing assay를 통해 EAA가 HaCaT 세포의 이주 능력에 영향을 주는지 관찰하였다. 또한 western blot analysis와 qRT-PCR을 통하여 EAA가 $TNF-{\alpha}/IFN-{\gamma}$로 자극한 HaCaT 세포에서 iNOS의 단백질 발현 및 IL-4, IL-13, IL-6의 mRNA 발현, filaggrin의 단백질과 mRNA 발현에 미치는 영향을 조사하였다. 결과 : EAA는 처리 농도 $500{\mu}g/ml$까지 HaCaT keratinocytes의 세포생존율에 영향을 미치지 않았다. EAA는 wound healing assay에서 HaCaT 세포의 이주 능력을 증가시켰으며, $TNF-{\alpha}/IFN-{\gamma}$로 자극한 HaCaT 세포에서 iNOS의 단백질 수준을 감소시켰다. 또한 EAA가 IL-4, IL-13, IL-6의 mRNA 발현을 억제하는 것 역시 확인할 수 있었다. 뿐만 아니라 EAA는 $TNF-{\alpha}/IFN-{\gamma}$ 자극에 의해 감소했던 filaggrin을 단백질과 mRNA 수준에서 회복시켰다. 결론 : EAA가 HaCaT 세포에서 Th2 type cytokines, pro-inflammatory cytokine의 억제와 filaggrin 회복을 통해 피부장벽 기능 손상에 대한 억제활성을 갖는 것을 확인하였으며, 이를 통해 EAA가 염증으로 인해 손상된 피부장벽 기능 개선에 효과적일 것으로 사료된다.

• Toxicological Research
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• 제34권1호
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• pp.7-12
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• 2018

Laboratory animal models have been developed to investigate preventive or therapeutic effect of medicinal products, or occurrence or progression mechanism of atopic dermatitis (AD), a pruritic and persistent inflammatory skin disease. The murine model with immunologic phenomena resembling human AD was introduced, which demonstrated skewedness toward predominance of type-2 helper T cell reactivity and pathophysiological changes similar as human AD following 2,4-dinitrochlorobenzene (DNCB) sensitization and challenge. Molecular mechanism on the DNCB-mediated AD was further evaluated. Skin tissues were collected from mice treated with DNCB, and each tissue was equally divided into two sections; one for protein and the other for mRNA analysis. Expression of filaggrin, an important protein for keratinocyte integrity, was evaluated through SDS-PAGE. Level of mRNA expression for cytokines was determined through semi-quantitative reverse transcriptase polymerase chain reaction. Expression of filaggrin protein was significantly enhanced in the mice treated with DNCB compared with the vehicle (acetone : olive oil = 4 : 1 mixture) treatment group or the normal group without any treatment. Level of tumor necrosis factor-alpha and interleukin-18 mRNA expression, cytokines involved in activity of type-1 helper T ($T_H1$) cell, was significantly downregulated in the AD group compared with other control groups. These results suggest that suppression of $T_H1$ cell-mediated immune response could be reflected into the skin tissue of mice treated with DNCB for AD induction, and disturbance of keratinocyte integrity might evoke a compensatory mechanism.

• 대한한의학회지
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• 제38권2호
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• pp.41-52
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• 2017

Objectives: Hataedock is indigenous treatment of Korean medicine that administers herbal extracts orally to newborn infants for remove the fetal heat. The purpose of this study was to evaluate skin lipid barrier formation effect of Hataedock treatment with Douchi. Methods: We measured the Western blot to observe the expression of protein such as involucrin and loricrin. Moreover, we observed immunohistochemical changes in NC/Nga mice. The 3-week-old NC/Nga mice were divided into 3 groups: the 3-week-old control group (3w-Ctrl), 5-week-old control group (5w-Ctrl), and the Hataedock-treated group (5w-FGT). Only the 5w-FGT group was treated with Douchi at the 3rd week. We identified the changes of the lipid skin barrier and protein differentiation through immunohistochemical changes of involucrin, loricrin, filaggrin and acid sphingomyelinase (ASM) in the stratum corneum. Results: The expression of involucrin and loricrin was increased in the Western blot that was treated with concentration of Douchi extracts. In 5w-FGT group, loricrin-positive reaction was increased by 54.0%, involucrin-positive reaction was increased by 84.0%, filaggrin-positive reaction was increased by 108.0% and ASM-positive reaction was increased by 91.0% in the stratum corneum. Conclusions: These results suggest that Hataedock treatment with Douchi promoted skin lipid barrier formation by promoting differentiation of keratinocytes.

• Journal of Ginseng Research
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• 제44권1호
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• pp.115-122
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• 2020

Background: In aged skin, degradation of collagen fibers, which occupy the majority of the extracellular matrix in the dermis, and changes of aquaporin 3 (AQP3) and skin constituents, such as hyaluronic acid and ceramide, cause wrinkles and decrease skin moisturization to contribute to dryness and lower elasticity skin. Red ginseng (RG) is used as a cosmetic and food material and is known to protect from UVB-induced cell death, increase skin hydration, prevent wrinkles, and have an antioxidative effect. But, in general, RG used as a material is the soluble liquid portion in the solvent, and the part that is not soluble in the solvent is discarded. Thus, we made the whole RG into microgranulation and dispersed in water to produce gel form for using entire RG, and it was named red ginseng NaturalGEL (RG NGEL). Methods: RG NGEL was investigated for matrix metalloproteinases inhibitory activity, induction of Type I collagen, AQP3, hyaluronan synthetase 2, serine palmitoyl transferase, ceramide synthase 3, and filaggrin expression and compared with RG water extract. Results: RG NGEL reduced the levels of UV-induced matrix metalloproteinases and increased Type I collagen in human fibroblast cells and upregulated AQP3, hyaluronan synthetase 2, serine palmitoyl transferase, ceramide synthase 3, and filaggrin expressions in human keratinocytes compared with RG water extract. Conclusion: RG NGEL has the potential as an effective reagent for antiaging cosmetics to improve wrinkle formation and skin hydration.

• 대한한방소아과학회지
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• 제34권4호
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• pp.22-30
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• 2020

Objectives This study was conducted to confirm the inhibitory effect of β-glucan on epithelial inflammation induced by atopic dermatitis through Endocannabinoid system (ECS) activity. Methods Six-week-old NC/Nga mice were divided into a control group (Ctrl), atopic dermatitis elicitation group (ADE), and a β-glucan-treated group (β-glucan treatment after atopy dermatitis elicitation, β-GT). After 3 weeks, CB1, CB2, and GPR55 were observed to confirm the regulation of ECS activity, and filaggrin in the stratum corneum and Kallikrein-related peptidase (KLK) 7 in the stratum corneum and protease activated receptor (PAR)-2 were observed to confirm the inhibition of the inflammation, Phosphorylated extracellular signal-related kinase (p-ERK), Phosphorylated mammalian target of rapamycin (p-mTOR), and E-Cadherin were observed to confirm microenvironmental regulation. Results β-GT was significantly increased in CB1, CB2, and GPR55 positive reactions compared to that of the ADE. In positive reaction of the filaggrin in the stratum corneum, β-GT was significantly increased than that of the ADE. For KLK7 positive and PAR2 positive, β-GT was significantly reduced compared to the ADE. The p-ERK-positive and p-mTOR-positive reactions were significantly reduced in β-GT than in ADE. E-cadherin positive reaction was significantly increased in β-GT than in ADE (All p < 0.01). Conclusions It was confirmed that β-glucan has the effect of inhibiting the epithelium induced by atopic dermatitis through the ECS activity.

• Toxicological Research
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• 제27권4호
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• pp.225-229
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• 2011

Extract of Taraxacum platycarpum (AF-343) has been reported to have several biological properties such as skin hydration and anti-inflammatory effects. Although clinical evidences of skin hydration and antiinflammatory effect were proven in clinical trial, precise mechanism of skin hydration was not fully understood yet. In this study, we have focused skin hydration mechanism related filaggrin, collagen, and matrix metalloproteinase (MMP) in vitro and animal study. Herein, skin hydration mechanism of AF-343 is due to recovery of filaggrin in mice model and increased production of collagen with suppression of matrix MMP in vitro fibroblast cell line.