• BMB Reports
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• v.51 no.4
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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.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.43 no.4
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• pp.329-335
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• 2017

We have studied on the keratinocytes differentiation and skin barrier function using Adenophorae radix (A. radix) root extract, which was known to contain triterpenoid, saponin and starch. A. radix root extracts showed the $PPAR{\alpha}$ expression level of Wy-14,643 $0.5-1.0{\mu}M$ in CV-1 cells. The cornified envelop formation (CE) of human keratinocyte cell line (HaCaT) and normal human keratinocyte (NHK) showed a statistically significant increased compared to the control. When HaCaT cells were treated with A. radix root extract, transglutaminase (TGase-1) was significantly increased. As a result of clinical study of the simple cosmetic formulation containing A. radix root extract for about 2 weeks, TEWL values were significantly decreased and water contents were increased. The ceramides, which were obtained from the inner forearm, were also significantly increased statistically. We suggest that the A. radix root extract can be used as a preventive and therapeutic agent for skin diseases such as dry skin and atopy.

• Journal of Physiology & Pathology in Korean Medicine
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• v.32 no.2
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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.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.35 no.2
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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.

• The Korea Journal of Herbology
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• v.32 no.3
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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.

• Journal of Physiology & Pathology in Korean Medicine
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• v.25 no.6
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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.

• Journal of Korean society of Dental Hygiene
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• v.15 no.4
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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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• v.48 no.9
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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]

• Proceedings of the Korean Society of Applied Pharmacology
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• 2003.11a
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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.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.42 no.4
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• pp.359-366
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• 2016

Exosome, a small vesicle secreted from cells, has diverse functions depending on cell origins and tissue types and plays a important role in cell viability and intercellular communication. Recently, many researchers have demonstrated the use of exosomes for the treatment of cancers and immune diseases, and the development of diagnostic biomarker. However, the secretion mechanism of exosome from skin cell and its physiological functions in skin remain unclear. Thus, this study aimed to explore whether keratinocyte-derived exosome affects proliferation and migration in HaCaTs. Exosomes were isolated from HaCaTs by ExoQuick-TC and then boiled or unbolied. Boiled and unboiled exosome induced proliferation in HaCaTs in a dose-dependant manner ($0.1{\sim}20{\mu}g/mL$), respectively. Boiled and unboiled exosome at concentration of $20{\mu}g/mL$ increased proliferation level in HaCaTs by $186.96{\pm}3.87%$ and $193.48{\pm}10.48%$ compared with control group. Unboiled exosome stimulated migration in HaCaTs in a dose-dependent manner ($0.1{\sim}20{\mu}g/mL$), which reached a maxium at concentration of $20{\mu}g/mL$ ($179.39{\pm}4.89%$ of control), but boiled exosome did not affect HaCaT migration. In addition, unboiled exosome ($0.1{\sim}20{\mu}g/mL$) dose-dependently stimulated sprout outgrowth in HaCats. These results demonstrate that in exosome from HaCaTs, heat-stable components such as lipid may induce HaCaT proliferation and heat-unstable components such as protein may stimulate migration and sprout outgrowth in HaCaTs, thereby leading to reepithelialization and skin-wound healing activities. It is concluded that exosomes from HaCaTs may be used as cosmetic materials.