• 동의생리병리학회지
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• 제32권4호
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• pp.226-231
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• 2018

Chijabaegpi-tang (CHG) is an oriental herbal medicine that has been used for its various pharmacological effects, which include anti-inflammatory, anti-oxidant and immunoregulation activities. In the present study, we investigated which skin inflammations are involved in the $TNF-{\alpha}/IFN{\gamma}$-induced HaCaT cells. We investigated the suppressive effect of CHG on $TNF-{\alpha}/IFN{\gamma}$-induced HaCaT cell production of the following chemokines: macrophage-derived chemokine (MDC)/CCL22; regulated on activation, normal T-cell expressed and secreted (RANTES)/CCL5; and interleukin-8 (IL-8); thymus and activation-regulated chemokine (TARC)/CCL17. The pre-treatment of HaCaT cells with CHG suppressed $TNF-{\alpha}/IFN{\gamma}$-induced nuclear transcription factor kappa-B ($NF-{\kappa}B$). In addition, CHG inhibited $TNF-{\alpha}/IFN{\gamma}$-induced phosphorylation of ERK and p38. $TNF-{\alpha}/IFN{\gamma}$ suppressed the expression of skin barrier proteins, including filaggrin (FLG), Involucrin (IVL) and loricrin (LOR). By contrast, CHG restored the expression of FLG, IVL and LOR. Taken together, our findings suggest that CHG could be a therapeutic agent for prevention of skin disease, including atopic dermatitis.

• 한국자원식물학회지
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• 제32권5호
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• pp.433-441
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• 2019

본 연구는 하라케케(Harakeke)로 불리는 신서란(Phormium tenax)를 화장품 및 의약품산업의 기능성 소재로서의 이용 가능성을 확인하기 위하여 신서란 잎을 대상으로 70% 에탄올 추출물과 용매 분획물을 제조하여, 이것들의 항염증 및 항아토피의 효과를 조사하였다. LPS로 유도된 RAW 264.7 세포에서 신서란 에탄올 추출물과 용매 분획물의 항염증 효과를 조사한 결과, methylene chloride와 ethyl acetate 분획물에서 NO와 $PGE_2$ 생성 억제 활성이 가장 높게 나타났으며, 농도 의존적으로 NO와 $PGE_2$ 생성 억제 활성을 보였다. 또한, 이들 분획물에서는 iNOS 및 COX-2 발현 억제 활성을 보였다. 신서란 잎 조추출물과 용매 분획물에 의한 NO, $PGE_2$ 생성 억제 활성이 NOS 및 COX-2 발현 억제에 의한 것임을 제시한다. 더불어, $hIFN-{\gamma}$로 자극된 HaCaT 세포에 용매 분획물을 처리하여 MDC 및 TRAC 생성억제 효과를 조사한 바, methylene chloride 분획물은 MDC 및 TATC의 생성을 각각 65%, 52% 생성억제 시켰으며, ethyl acetate 분획물은 MDC 및 TATC의 생성을 각각93%, 84% 억제 효과를 보였다. 이상의 결과는 신서란 잎 조추출물과 용매 분획물을 이용한 항염증 및 항아토피 효능을 갖는 유효성분 분리 및 활용화 연구에 중요한 기초자료가 될 것이며, 기능성 화장품, 의약외품 및 의약품 소재 개발에 적용 가능성이 높다고 사료된다.

• 대한본초학회지
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• 제34권1호
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• pp.23-31
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• 2019

Objectives : The aim of this study was to investigate the effect for allergic-inflammation of Fritillariae Thunbergii Bulbus (FTB) on HaCaT cells and RBL2H3 cells. Methods : To investigate the effects of FTB for anti-inflammation in HaCaT cells, the cells were pretreated with FTB for 1h and then stimulated with $TNF-{\alpha}/IFN-{\beta}$ for 24h. Then thymus and activation-regulated chemokine (TARC) and Macrophage-derived chemokine (MDC) levels were analyzed with ELISA kit. Also to investigate the effect of skin barrier protein, the cells were treated with FTB of various concentrations, and then cells were harvested, expressions of skin barrier protein were measured with RT-PCR. To investigate the effects of FTB for anti-allergy in RBL2H3 cells, the cells were pre-treated with FTB for 1h, and then stimulated with A23187 for 30 min. ${\beta}$-hexosaminidase, IL-4 and $TNF-{\alpha}$ were measured using cultured media. The cells were harvested to analyze the mechanism of the effect for FTB via Western blot. Results : FTB did not show cytotoxicity in HaCaT and RBL2H3. In HaCaT cells, FTB significantly suppressed the expression of TARC, MDC at a dose-dependent manner and markedly increased formation of the skin barrier proteins. In RBL2H3 cells, FTB decreased release of the ${\beta}$-hexosaminidase, IL-4 and $TNF-{\alpha}$ in RBL2H3 through inhibition of the phosphorylation of JNK and p38, which are include in the signaling mechanism of MAPK Conclusion : These results indicate that FTB has an anti-inflammatory effect on the allergic response through blocking MAPK pathway. This suggest that FTB could be a therapeutic agent for allergic response.

• 대한본초학회지
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• 제37권3호
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• pp.9-19
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• 2022

Objectives : Angelicae Gigantis Radix (AG) is a plant of the Ranunculus family. AG have been reported to have various pharmacological effects on human health which include uterine growth promotion, anti-inflammatory, analgesic, and immune enhancement. However, research on dermatitis disease is insufficient. Therefore, we investigated the effects of AG on tumor necrosis factor-α (TNF-α)/interferon-γ (IFN-γ) stimulated HaCaT cell. Methods : To investigate the effect of AG on HaCaT cell, HaCaT cells were pre-treated with AG for 1 hour and then stimulated with TNF-α/IFN-γ. After 24 hours, media and cells were harvested to analyze the inflammatory mediators. Concentration of human interleukin-1beta (IL-1β), monocyte chemoattractant protein-1 (MCP-1), granulocyte-macrophage colony-stimulating factor (GM-CSF), and TNF-α in the media were assessed by ELISA. mRNA expression of human thymus and activation-regulated chemokine (TARC), IL-6, and IL-8 were analyzed by RT-PCR. Additionally, the mechanisms of mitogen-activated protein kinases (MAPKs) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathway were investigated by Western blot. Results : The treatment of AG inhibited gene expression levels of IL-6, IL-8, and TARC and protein expression levels of IL-1β, MCP-1, and GM-CSF. Also, AG significantly reduced extracellular signal-regulated kinase (ERK) phosphorylation and NF-κB translocation in TNF-α/IFN-γ stimulated HaCaT cell. Conclusions : Taken together, these results demonstrate that AG can alleviate inflammatory diseases such as atopic dermatitis by regulating the expression of inflammatory cytokines. Also, it suggest that AG may a promising candidate drug for the treatment of inflammatory disease such as atopic dermatitis.

• 대한본초학회지
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• 제35권4호
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• pp.51-60
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• 2020

Objective : The objective of this study was to demonstrate the effect of Persicae Semen (PS) in DNCB-induced atopic dermatitis mouse and HaCaT cell. Methods : The BALB/c mice were divided into four groups. To develop atopic dermatitis, 200 ㎕ of 1 and 0.5% DNCB solution was put on the back of mice in the Control group, the PS-Low group and the PS-High group once a day. After application of DNCB, 200 ㎕ of the PS extract was also treated. The Normal group was given PBS. The mice dorsal skin was stained with Masson's trichrome, H&E, and toluidine blue to evaluate the thickness of the epidermis and dermis, infiltration of eosinophils and mast cells respectively. ELISA was applied to measure the serum level of IgE and IL-6. Toxicity of PS was measured by MTS assay in HaCaT cell. To investigate the effects of PS on HaCaT cells, cells were pre-treated with PS for 1h, and then stimulated with TNF-α and IFN-γ. After 24 hours, the expression of TARC was analyzed using RT-PCR. Results : PS not only significantly diminished the thickness of the epidermis and dermis, but also reduced the infiltration of eosinophil and mast cell in skin lesion. PS also reduced the serum IgE and IL-6 level which plated important roles in the atopic dermatitis. The expression of TARC was decreased significantly in TNF-α/IFN-γ stimulated HaCaT cell. Conclusion : These results suggest that PS may be effective in alleviating the atopic dermatitis induced by DNCB and inflammation by TNF-α/IFN-γ.

• Biomolecules & Therapeutics
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• 제24권5호
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• pp.529-535
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• 2016

Atopic dermatitis (AD) results from gene and environment interactions that lead to a range of immunological abnormalities and breakdown of the skin barrier. Protease-activated receptor 2 (PAR2) belongs to a family of G-protein coupled receptors and is expressed in suprabasal layers of the epidermis. PAR2 is activated by both trypsin and a specific agonist peptide, SLIGKV-$NH_2$ and is involved in both epidermal permeability barrier homeostasis and epithelial inflammation. In this study, we investigated the effect of lobaric acid on inflammation, keratinocyte differentiation, and recovery of the skin barrier in hairless mice. Lobaric acid blocked trypsin-induced and SLIGKV-$NH_2$-induced PAR2 activation resulting in decreased mobilization of intracellular $Ca^{2+}$ in HaCaT keratinocytes. Lobaric acid reduced expression of interleukin-8 induced by SLIGKV-$NH_2$ and thymus and activation regulated chemokine (TARC) induced by tumor necrosis factor-a (TNF-${\alpha}$) and IFN-${\gamma}$ in HaCaT keratinocytes. Lobaric acid also blocked SLIGKV-$NH_2$-induced activation of ERK, which is a downstream signal of PAR2 in normal human keratinocytes (NHEKs). Treatment with SLIGKV-$NH_2$ downregulated expression of involucrin, a differentiation marker protein in HaCaT keratinocytes, and upregulated expression of involucrin, transglutamase1 and filaggrin in NHEKs. However, lobaric acid antagonized the effect of SLIGKV-$NH_2$ in HaCaT keratinocytes and NHEKs. Topical application of lobaric acid accelerated barrier recovery kinetics in a SKH-1 hairless mouse model. These results suggested that lobaric acid is a PAR2 antagonist and could be a possible therapeutic agent for atopic dermatitis.

• Journal of Applied Biological Chemistry
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• 제58권1호
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• pp.13-19
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• 2015

본 연구에서는 우슬 열수 추출물을 n-BuOH과 $H_2O$로 용매 분획 한 후 n-BuOH 분획물을 컬럼 크로마토그래피를 수행하여 6개의 소분획물(Fr. 1-6)을 얻었다. 소분획물 6개에 대해 tumor necrosis factor (TNF)-${\alpha}$ 저해활성 검정을 통하여 가장 높은 활성을 나타낸 분획물 5에서 preparative-high performance liquid chromatography를 이용해 3종의 phytoecdysone계열 화합물들을 분리하였다. 분리된 화합물들은 NMR 및 MS 분석을 통해 ecdysterone (1), 25S-inokosteron (2), 25R-inokosteron (3)임을 동정 하였고, 이들의 활성은 TNF-${\alpha}$ 생산 저해활성은 $200{\mu}g/mL$에서 80-95%의 TNF-${\alpha}$ 저해활성을 나타내었다. 또한 각질형성 세포에 분리한 3종의 화합물을 처리한 결과 각각 51와 23% 이상의 TARC의 생성 억제 효과를 확인 할 수 있었다. 이들의 결과로써, 우슬 추출물과 분리된 화합물들에 대한 항 아토피 활성 결과로 천연물 기능성 식품 또는 화장품으로의 응용가능성이 높다고 사료된다.

• 한국식품영양과학회지
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• 제46권3호
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• pp.298-305
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• 2017

본 연구는 DNCB로 유도한 아토피 피부염 유사 병변 실험동물 모델을 활용한 in vivo와 피부 각질세포주를 이용한 in vitro 연구를 통하여 아로니아와 모링가 종자 추출물의 아토피 피부염 개선 효능을 분석하였다. 조직학적 검사 결과 아로니아 추출물(AM) 또는 모링가 종자 추출물(MO) 단독 투여 시보다 1:1로 혼합한 복합물(AM-MO)을 투여했을 때 표피 부분의 과증식과 염증세포의 침윤이 감소하였으며, 염증 및 부종을 나타내는 피부 두께를 측정한 결과에서도 추출물 단독으로 투여하였을 때보다 AM-MO 투여 시 DNCB 도포군과 비교하여 유의하게 감소함을 보여주었다. 피부장벽 손상으로 인한 수분 손실에 대해서는 유의한 효과를 나타내지 않았으며, 혈청 IgE 수준은 AM과 MO 각각 단독으로 투여했을 때보다 혼합 투여하였을 때 현저하게 억제되었다. In vitro에서는 피부각질세포주인 HaCaT 세포를 $TNF-{\alpha}$ 및 $IFN-{\gamma}$로 자극하면 발현이 증가하는 Th2 케모카인 TARC 및 MDC에 대하여 mRNA 수준을 분석하였을 때, 이 역시 단독 추출물 처리 시보다 복합물 처리 시 억제 효과가 더 뛰어남을 확인하였다. 종합적으로 DNCB로 유도된 아토피 피부염 in vivo 모델에서 아로니아 및 모링가 종자 추출물은 단독으로 투여하는 것보다 복합물로 투여하였을 때 현저하게 피부 염증 및 혈청 내 IgE 생성을 억제하였으며, 피부각질 세포주를 이용한 in vitro 연구에서도 추출물 단독 처리보다 혼합하여 처리하였을 때 더 유의한 Th2 케모카인 억제 활성을 나타내었다. 이로 인해 아로니아 및 모링가 종자 추출물은 복합적으로 사용하였을 때 상호작용에 의해서 더 효과적인 아토피 피부염 개선 식품 소재로써 활용될 가능성이 높을 것으로 생각된다.