• BMB Reports
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• 제51권4호
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• pp.188-193
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• 2018

Caffeoylserotonin (CaS), one derivative of serotonin (5-HT), is a secondary metabolite produced in pepper fruits with strong antioxidant activities. In this study, we investigated the effect of CaS on proliferation and migration of human keratinocyte HaCaT cells compared to that of 5-HT. CaS enhanced keratinocyte proliferation even under serum deficient condition. This effect of CaS was mediated by serotonin 2B receptor (5-HT2BR) related to the cell proliferation effect of 5-HT. We also confirmed that both CaS and 5-HT induced G1 progression via 5-HT2BR/ERK pathway in HaCaT cells. However, Akt pathway was additionally involved in upregulated expression levels of cyclin D1 and cyclin E induced by CaS by activating 5-HT2BR. Moreover, CaS and 5-HT induced cell migration in HaCaT cells via 5-HT2BR. However, 5-HT regulated cell migration only through ERK/AP-1/MMP9 pathway while additional Akt/NF-${\kappa}B$/MMP9 pathway was involved in the cell migration effect of CaS. These results suggest that CaS can enhance keratinocyte proliferation and migration. It might have potential as a reagent beneficial for wound closing and cell regeneration.

• 대한화장품학회지
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• 제43권4호
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• pp.329-335
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• 2017

Triterpenoid, saponin, 전분 등이 함유되어 있는 것으로 알려진 Adenophorae radix (A. radix)의 뿌리 추출물을 이용하여 각질형성세포의 분화와 피부장벽기능에 대한 연구를 수행하였다. A. radix의 뿌리 추출물은 CV-1 세포를 이용하여 $PPAR{\alpha}$ 발현을 살펴본 결과, Wy-14,643 $0.5-1.0{\mu}M$ 수준의 발현양을 나타내었다. 인체 각질형성 세포주(HaCaT)와 각질형성세포(nomal human keratinocyte)에 대한 각질형성능(cornified envelop formation, CE)은 대조군에 비해 통계적으로 유의한 증가를 보였다. HaCaT 세포에 A. radix의 뿌리 추출물 처리하였을 때, transglutaminase (TGase-1)의 유의적 증가를 보였다. A. radix의 뿌리 추출물을 함유한 간단한 화장품 제형을 약 2주간에 걸쳐 임상시험을 실시한 결과, TEWL의 유의적 감소와 수분량의 증가를 살펴볼 수 있었으며, 하박 내측에서 지질을 추출하여 세라마이드를 분석한 결과 통계적으로 유의한 증가를 관찰할 수 있었다. 이를 통하여 A. radix의 뿌리 추출물을 건조피부나 아토피 등의 피부질환과 관련된 질환의 예방 및 치료제로 사용될 수 있을 것이다.

• 동의생리병리학회지
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• 제32권2호
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• pp.106-112
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• 2018

The aim of this research was to determine the diverse effects of Sappan Lignum extract and brazilin on human keratinocyte HaCaT cells. We confirmed the antioxidant effect of Sappan Lignum extract and brazilin was analyzed by using an ABTS assay, confirming the efficacy of water extraction method. Also, we examined effect of Sappan Lignum extract and brazilin on the cell viability, using the MTS assay in HaCaT cells. mRNA expression levels of tight junction-related genes associated with skin barrier in HaCaT cells were analyzed using quantitative real-time PCR analysis. Sappan Lignum extract increased the cellular activity of HaCaT cells and the expression of the tight junction-related genes claudin 3, claudin 6, and ZO-2. Brazilin displayed the same effects as that of the extract on HaCaT cells activity and tight junction-related genes expression. Furthermore, dispase assay demonstrated altered cell-cell adhesion strength of Sappan Lignum extract or brazilin treated HaCaT cells. Sappan Lignum extract or brazilin might be an useful ingredient in skin-mosturizinng and anti-wrinkle cosmetics, given its effects of altering mRNA expression of tight junction-related genes and enhancing cell-cell adhesion strength of HaCaT cells.

• 한방안이비인후피부과학회지
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• 제35권2호
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• pp.13-27
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• 2022

목적 : 본 연구는 인간피부각질형성세포(HaCaT keratinocytes) 모델을 TNF-α와 IFN-γ로 자극하여 내복자(萊菔子)의 피부염증 감소 및 만성 염증성 질환에 미치는 영향을 알아보고자 하였다. 방법 : 내복자(萊菔子) 에탄올 추출물(RSE)이 세포생존율에 미치는 영향을 확인하기 위하여 MTT assay를 시행하였다. 또한 RSE가 TNF-α와 IFN-γ로 자극한 HaCaT 세포에서 p-IκBα, p-ERK, p-JAK2, p-STAT1, p-STAT6의 발현과 periostin, TSLP 단백질 발현에 미치는 영향을 확인하였다. 결과 : RSE는 200㎍/㎖ 이하에서 세포 독성을 보이지 않았고, HaCaT keratinocytes에서 TNF-α와 IFN-γ자극에 의하여 증가된 IκBα, ERK의 인산화를 억제하였다. 또한 JAK2와 STAT1, STAT6의 인산화를 억제하였으며, periostin과 TSLP의 발현을 감소시켰다. 결론 : RSE는 HaCaT keratinocytes에서 pro-inflammatory cytokines 및 transcription factors의 발현을 감소시켜 피부염증 감소 효능을 보였고, 만성 염증성 질환에서 내복자(萊菔子)의 사용 가능성을 확인하였다.

• 대한본초학회지
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• 제32권3호
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• pp.55-61
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• 2017

Objectives : The aim of this research was to determine the diverse effects of Lithospermi Radix Water Extract (LR) on human keratinocyte HaCaT cells, and to examine whether those effects could be applied to the human skin. Methods : We examined effect of LR on the cell viability of using the MTS assay in human keratinocyte HaCaT cells. The antioxidation effect of LR was analyzed relative to the well-known antioxidant resveratrol, using an ABTS assay. Quantitative RT-PCR analysis revealed that, in HaCaT cells, LR influenced the mRNA expression of tight-junction genes associated with skin moisturization. Furthermore, a wound-healing assay demonstrated altered cell migration in LR-treated HaCaT cells. Result : The cytotoxicity was confirmed to be higher in LR at a concentration of $800{\mu}g/m{\ell}$ using the MTS assay in HaCaT cells. In comparison to $100{\mu}M$ resveratrol, $1,600{\mu}g/m{\ell}$ LR showed either a similar or superior antioxidation effect. LR treatment in HaCaT cells reduced the mRNA expression levels of claudin 3, claudin 4, claudin 6, claudin 8, and ZO-2 to less than 0.80-fold, whereas JAM-A and Tricellulin mRNA expression level increased more than 1.33-fold. In addition, HaCaT cells migration was decreased to 83.9% by LR treatment. Conclusions : LR of antioxidation activity will have an anti-aging effect on the skin by reducing oxidative stress. Further studies are required to address the implications for human skin, given LR's effects of altering mRNA expression of tight junction-related gene and decreasing cell migration of HaCaT cells.

• 동의생리병리학회지
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• 제25권6호
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• pp.1014-1019
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• 2011

Curcumin is a natural phytochemical present in turmeric, the ground powder of the rhizomes of Curcuma longa. Curcumin has been described as having antioxidant, anti-inflammatory, and anti-carcinogenic properties. However, it is still largely unknown whether curcumin inhibits the UVB-induced inflammatory reaction in HaCaT keratinocyte cell lines. In this study, to confirm the photoprotection properties of curcumin, HaCaT keratinocyte cells were irradiated by UVB, and then treated with curcumin. UVB irradiation induced the increased expressions of IL-$1{\beta}$, TNF-${\alpha}$, IL-6, IL-8, and COX-2 in HaCaT cells. These increased expressions of cytokines (IL-$1{\beta}$, TNF-${\alpha}$, IL-6, IL-8, and COX-2) were down-regulated by curcumin treatment in UVB-irradiated HaCaT cells. In addition skin of hairless mice were damaged by UVB irradiation, which were evidenced by atrophy of epidermis and decrease of collagen in dermis. However, these damages were protected partially by co-treatment of curcumin. Taken together, this data indicate that curcumin may be a promising photoprotection agent, when used in massage pack or sunscreen product, to reduce cell death in UV-damaged skin.

• 한국치위생학회지
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• 제15권4호
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• pp.719-725
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• 2015

Objectives: The purpose of this study was to determine the toxic effects of sodium lauryl sulfate(SLS) in human keratinocyte HaCaT cells and mouse fibroblast NIH-3T3 cells. Methods: The effect of sodium lauryl sulfate(SLS) cell viability and proliferation were determined by WST-1 assay and changes shape of nucleus were evaluated by Hoechst staining under fluorescence microscopy. Additionally, observation of cell morphological changes under light microscopy. Results: SLS induced cytotoxicity and a marked apoptosis in both HaCaT and NIH-3T3 cell lines. With the result of the WST-1 assay, SLS induced the cytotoxicity of 0.005% and 0.0075%, 0.01% SLS for 24 h after HaCaT and NIH-3T3 cells in time and dose-dependent manner(p<0.005). SLS inhibited cell growth and caused apoptosis as evidenced by nuclear fragmentation and condensation. Thus, determination of the morphological changes to define apoptosis was visualized using inverted phase contrast microscopy. Conclusions: SLS had toxicity of the human keratinocyte cells and mouse fibroblast cells and this study will provide the basic data for the development of proper SLS concentration in dentifrice.

• BMB Reports
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• 제48권9호
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• pp.495-500
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• 2015

Up-regulation of cell adhesion molecules and proinflammatory cytokines contributes to enhanced monocyte adhesiveness and infiltration into the skin, during the pathogenesis of various inflammatory skin diseases, including atopic dermatitis. In this study, we examined the anti-inflammatory effects of butein, a tetrahydroxychalcone, and its action mechanisms using TNF-α-stimulated keratinocytes. Butein significantly inhibited TNF-α-induced ICAM-I expression and monocyte adhesion in human keratinocyte cell line HaCaT. Butein also decreased TNF-α-induced pro-inflammatory mediators, such as IL-6, IP-10 and MCP-1, in HaCaT cells. Butein decreased TNF-α-induced ROS generation in a dose-dependent manner in HaCaT cells. In addition, treatment of HaCaT cells with butein suppressed TNF-α-induced MAPK activation. Furthermore, butein suppressed TNF-α-induced NF-kappaB activation. Overall, our results indicate that butein has immunomodulatory activities by inhibiting expression of proinflammatory mediators in keratinocytes. Therefore, butein may be used as a therapeutic agent for the treatment of inflammatory skin diseases. [BMB Reports 2015; 48(9): 495-500]

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2003년도 Annual Meeting of KSAP : International Symposium on Pharmaceutical and Biomedical Sciences on Obesity
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• pp.65-65
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• 2003

The aim of this study was to investigate the effect of resveratrol on the oxidative stress-induced inhibition of gap junctional intercellular communication (GJIC) in HaCaT keratinocyte. Anti-oxidative activity of resveratrol was measured by a,a-diphenyl-b-picrylhydrazyl (DPPH) assay and dichlorodihydrofluorescein diacetate oxidation assay. GJIC of HaCaT keratinocyte was assessed using the scrape loading/dye transfer technique. Western blots and reverse transcription-polymerase chain reaction were also analyzed for Connexin 43 protein and mRNA expression, respectively. Resveratrol scavenged directly the stable DPPH radical over a concentration range of 4 mg/$\mell$ (78.2 $\pm$ 2.7% of control) to 500 mg/$\mell$ (29.9 $\pm$ 4.2% of control) and prevent to increase the intracellular fluorescence induced by oxidative stress significantly. Ultraviolet A irradiation (UVA) and 12-O-tetradecanoylphorbol-13-acetate markedly reduced GJIC, which was restored by resveratrol. There were no significant differences in the level of Connexin 43 protein and mRNA expression among any of the experimental groups. Our data suggests that resveratrol has the protective effect on the oxidative stress-induced inhibition of gap junctional intercellular communication in HaCaT keratinocyte and this protection is likely due to the scavenging of reactive oxygen species.

• 대한화장품학회지
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• 제42권4호
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• pp.359-366
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• 2016

엑소좀은 세포에서 분비되는 작은 소낭체로서, 기원세포와 조직에 따라 다양한 기능을 수행하며, 세포생존 및 세포 간 커뮤니케이션에 중요한 역할을 한다. 최근 엑소좀을 활용하여 종양연구, 면역질환 개선, 질병진단 bio-marker 개발 등 다양한 분야에서 연구가 진행되고 있으나, 피부세포에서 분비기전 및 피부 생리적 기능에 대한 연구는 미흡한 실정이다. 따라서 본 연구에서 인체피부 유래 각질형성세포(HaCaT)로부터 분리된 엑소좀이 피부각질형성세포의 증식과 이주에 미치는 영향을 확인하고자 하였다. HaCaT으로부터 ExoQuick-TC를 활용하여 엑소좀을 분리하고, 열처리 엑소좀(boiled exosome)과 무처리 엑소좀(unboiled exosome)으로 구분하였다. HaCaT 유래 엑소좀은 농도 의존적으로($0.1{\sim}20{\mu}g/mL$) HaCaT의 증식을 유도하였으며, $20{\mu}g/mL$에서 대조군(control)에 비해 각각 $186.96{\pm}3.87%$(열처리) 또는 $193.48{\pm}10.48%$(무처리)의 증식 유도활성을 나타내었다. 또한 HaCaT 유래 무처리 엑소좀은 농도 의존적인($0.1{\sim}20{\mu}g/mL$) HaCaT의 이주활성을 나타내었고, $20{\mu}g/mL$에서 대조군에 비해 $179.39{\pm}4.89%$의 이주를 유도하였다. 그러나 열처리 엑소좀은 이주 유도활성을 나타내지 않았다. 뿐만 아니라 무처리 HaCaT 유래 엑소좀은 collagen sprout outgrowth를 농도 의존적으로 유도함을 확인하였다. 이러한 결과를 통해서 HaCaT 유래 엑소좀은 지질 및 열에 안정한 물질이 세포의 증식을 유도하고, 단백질 또는 열에 불안정한 물질이 세포의 이동 및 sprout out growth 활성에 관여하는 것으로 확인되었다. 따라서 피부각질형성세포 유래 엑소좀은 피부의 재상피화 및 상처치유 등의 활성을 나타낼 수 있으며, 향후 화장품소재로서 응용 가능성이 확인되었다.