• Journal of Society of Preventive Korean Medicine
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• 제22권2호
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• pp.109-123
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• 2018

Purpose : In this study, we intended to observe the anti-wrinkle and moisturizing effects of dried pomegranate juice concentration powder (PCP) using in vitro test. Materials and methods : Antioxidant effects of PCP were determined by free radical scavenging capacity (DPPH assay) and the cytotoxicity of PCP was examined in human keratinocyte (HaCaT) and human primary dermal fibroblast-neonatal (HDF) cells. To investigate the moisturizing effect of PCP, hyaluronan synthesis was examined in HaCaT cells. Activity of procollagen production were assessed in HDF cells and elastase inhibition properties of PCP were evaluated in cell free condition, to determine their anti-wrinkle effects. Metalloproteinase 1 (MMP-1) activity was also assessed following UVB irradiation, in the current in vitro experiment. Results : No PCP treatment related significant cytotoxic effects were demonstrated against to the both HDF and HaCaT cells. PCP showed favorable free radical scavenging activities in dose-dependent manner. In PCP-treated HaCaT cells, hyaluronan synthesis was non-significantly but markedly increased, and pro-collagen productions were significantly increased in HDF cells, at all three different concentrations (0.25, 0.75 and 1 mg/ml), and elastase inhibitory activities were observed by PCP treatment. A significant decrease in UVB-induced MMP-1 activity was also observed in 1 mg/ml PCP-treated HDF cells as compared to those of UVB-exposed cells. Conclusions : Taken together, these results suggest that PCP has favorable antioxidant, anti-wrinkle and moisturizing effects without meaningful cytotoxicity on HDF and HaCaT cell lines.

• Korean Journal of Acupuncture
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• 제34권2호
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• pp.82-87
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• 2017

Objectives : Recently the case of lactobacillus mixture culture fluid appliment was reported. In this study, anti-inflamation effects and anti-allergy effects were studied by stimulus of lactobacillus mixture culture fluid extracts in HaCaT cells. Methods : The atopic dermatitis were induced by TNF-${\alpha}$ and interferon-${\gamma}$ in HaCaT cells. TARC/CCL17, MDC/CCL22, RANTES/CCL5 and ROS production were investigated to explain anti-inflamation and allergy effects of lactobacillus mixture culture fluid with cell-enzyme-linked Immunosorbent assay in 450 nm, 485 nm, 535 nm with spectro-fluorometer. Results : The extracts of lactobacillus mixture culture fluid were decreased TARC/CCL17, MDC/CCL22, RANTES/CCL5 expressions and ROS production with a concentration dependent manner. Conclusions : The effects mechanism of Lactobacillus mixed culture fluid for atopic dermatitis symptoms were considered to be explain anti-inflamation and allergy effects via control of cytokine, chemokine and ROS production, and the fluid could be applied in skin cells directly. But classified AD symptom degrees reported in clinical case before as Reaction Period, Reduction Period, Effect Period, Reproduction Period and Rebound Period could not be explained. Further study will be expected.

• Korean Journal of Pharmacognosy
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• 제43권3호
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• pp.217-223
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• 2012

In the present study, we investigated anti-inflammatory activity of artemisinic acid in HaCaT cells and RAW264.7 cells. Artemisinic acid showed inhibitory activity on macrophage-derived chemokines (MDC) expression, a factor related with atopic dermatitis (AD), in interferon (IFN)-${\gamma}$ and tumor necrosis factor (TNF)-${\alpha}$-stimulated HaCaT cells. In the study on action mechanism, pretreated artemisinic acid reduced the phosphorylation of STAT1 and p38 and the degradation of $I{\kappa}B$ by IFN-${\gamma}$ and TNF-${\alpha}$ stimulations. However, artemisinic acid didn't show the inhibitory activity on LPS-induced inflammatory mediators (NO, $PGE_2$, IL-6) in RAW264.7 cell. These results indicate that artemisinic acid inhibits IFN-${\gamma}$ and TNF-${\alpha}$-induced MDC expression through inhibition of signal factors, STAT1, NF-${\kappa}B$, and p38, in HaCaT keratinocytes.

• Proceedings of the PSK Conference
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• 대한약학회 2004년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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• pp.103-103
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• 2004

• The Korea Journal of Herbology
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• 제34권2호
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• pp.25-32
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• 2019

Objectives : Gardeniae Fructus extract is used as a component of various cosmetics. However, the effect of the extract on the motility of keratinocytes has not been studied. The aim of this study is to investigate the inhibitory effect of ethanol extract of Gardeniae Fructus (GFET) or ethyl acetate extract of Gardeniae Fructus (GFEA) on oxidation and motility of human keratinocyte HaCaT cells. Methods : Antioxidant activity of Gardeniae Fructus extracts were determined by the 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) assay. To investigate the cytotoxicity of Gardeniae Fructus extracts, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was performed. The mRNA expression levels of tight junction related genes were analyzed using quantitative RT-PCR analysis. Cell migration assay was employed to determine the activity of Gardeniae Fructus extracts on motility of human keratinocyte HaCaT cells. Results : GFET and GFEA showed strong antioxidant activity. GFEA showed stronger cytotoxicity in HaCaT cells than GFET until $2.0mg/m{\ell}$ concentration. Cell migration assay demonstrated that GFET and GFEA decreased the motility of HaCaT cells. In addition, the mRNA expression level of claudin 8 among tight junction genes was significantly reduced by GFET or GFEA treatment. Conclusions : We investigated the physiological activities of the extracts of Gardeniae Fructus extracts on human keratinocytes by two different extraction methods. In addition, the mRNA expression level of claudin 8 among tight junction genes was significantly reduced by either GFET or GFEA treatment. This study provides basic information on the application of Gardeniae Fructus extract to cosmetics component.

• The Korea Journal of Herbology
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• 제38권5호
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• pp.61-67
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• 2023

Objective : Mori Fructus has been used to relieve thirst, and treat dizziness, tinnitus, and insomnia caused by poor constitution. This study was performed to investigate the protective effect of the ethanol extract of Mori Fructus (MF) in ultraviolet B (UVB)-induced apoptotic cell death in human keratinocyte cells. Methods : MF was prepared by extracting 100 g of Mori Fructus in 1 L of 100% ethanol for 48 h. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to measure cell viability. Apoptosis was determined by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, and the expression of apoptosis-related proteins was observed by Western blot. Tyrosinase activity was measured with a colorimetric commercial kit. Results : MF promoted cell vitality and inhibited apoptosis of UVB-induced HaCaT cells. MF pretreatment reduced TUNEL-positive cells and increased the expression of caspase-3 and -9. MF also displayed antioxidant effect with high radical scavenging ability. At 2 ㎎/㎖ concentration, the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical inhibition rates were 55.3 ± 4.6% and 48.5 ± 1.3%, respectively. Furthermore, MF showed a concentration-dependent inhibitory effect on tyrosinase. Conclusion : These results suggest that MF functions as a protective regulator in UVB-induced HaCaT cells by regulating apoptosis and partially exerting antioxidant effects. In addition, the tyrosinase inhibitory effect of MF shows the potential for MF to be used for skin pigmentation.

• Nutrition Research and Practice
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• 제16권1호
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• pp.1-13
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• 2022

BACKGROUND/OBJECTIVES: Bitter taste receptors are taste signaling pathway mediators, and are also expressed and function in extra-gustatory organs. Skin aging affects the quality of life and may lead to medical issues. The purpose of this study was to better understand the anti-skin aging effects of bitter taste receptors in D-galactose (D-gal)-induced aged human keratinocytes, HaCaT cells. MATERIALS/METHODS: Expressions of bitter taste receptors in HaCaT cells and mouse skin tissues were examined by polymerase chain reaction assay. Bitter taste receptor was overexpressed in HaCaT cells, and D-gal was treated to induce aging. We examined the effects of bitter taste receptors on aging by using β-galactosidase assay, wound healing assay, and Western blot assay. RESULTS: TAS2R16 and TAS2R10 were expressed in HaCaT cells and were upregulated by D-gal treatment. TAS2R16 exerted protective effects against skin aging by regulating p53 and p21, antioxidant enzymes, the SIRT1/mechanistic target of rapamycin pathway, cell migration, and epithelial-mesenchymal transition markers. TAS2R10 was further examined to confirm a role of TAS2R16 in cellular senescence and wound healing in D-gal-induced aged HaCaT cells. CONCLUSIONS: Our results suggest a novel potential preventive role of these receptors on skin aging by regulating cellular senescence and wound healing in human keratinocyte, HaCaT.

• International Journal of Oral Biology
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• 제39권2호
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• pp.97-105
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• 2014

The aim of this study was to determine the beneficial effect of propofol on human keratinocytes that have undergone hypoxia reoxygenation (H/R) injury and to investigate whether autophagy is associated with the protective mechanism. Thus, we evaluated how propofol influences the intracellular autophagy and apoptosis during the H/R process in the HaCaT cells. The cultured human keratinocyte cells were exposed to 24 h of hypoxia (5% $CO_2$, 1% $O_2$, 94% $N_2$) followed by 12 h of reoxygenation (5% $CO_2$, 21% $O_2$, 74% $N_2$). The experiment was divided into 4 groups: (1) Control=Normoxia ; (2) H/R=Hypoxia Reoxygenation ; (3) PPC+H/R=Propofol Preconditioning+Hypoxia Reoxygenation; (4) 3-MA+PPC+ H/R=3-MA-Methyladenine+Propofol Preconditioning+ Hypoxia Reoxygenation. In addition, Western blot analysis was performed to identify the expression of apoptotic pathway parameters, including Bcl-2, Bax, and caspase 3 involved in mitochondrial-dependent pathway. Autophagy was determined by fluorescence microscopy, MDC staining, AO staining, and western blot. The H/R produced dramatic injuries in keratinocyte cells. In our study, the viability of Propofol in H/R induced HaCaT cells was first studied by MTT assay. The treatment with 25, 50, and $100{\mu}M$ Propofol in H/R induced HaCaT cells enhanced cell viability in a dose-dependent manner and $100{\mu}M$ was the most effective dose. The Atg5, Becline-1, LC3-II, and p62 were elevated in PPC group cells, but H/R-induced group showed significant reduction in HaCaT cells. The Atg5 were increased when autophagy was induced by Propofol, and they were decreased when autophagy was suppressed by 3-MA. These data provided evidence that propofol preconditioning induced autophagy and reduced apoptotic cell death in an H/R model of HaCaT cells, which was in agreement with autophagy playing a very important role in cell protection.

• Journal of the Korean Applied Science and Technology
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• 제40권2호
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• pp.258-267
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• 2023

In this study, to investigate the possibility of using ethanol extract of Petasites japonicus (PJE) as a functional material, we investigated the activity of improving skin barrier and inflammation through UVB-induced human keratinocyte (HaCaT cell). As a result of confirming the antioxidant effect through DPPH radical scavenging activity, ABTS⁺ radical scavenging activity, and hydrogen peroxide scavenging activity, it was confirmed that it had an antioxidant effect similar to that of ascorbic acid, a control, at a concentration of 1 mg/ml. As a result of confirming the mRNA expression of the production ability of filaggrin and aquaporin-3 in HaCaT cells induced by UVB, it was confirmed that the reduced expression level by UVB stimulation increased in a concentration-dependent manner when the PJE was treated. It was confirmed that the mRNA expression of TNF-𝛼 and IL-1𝛽 were increased by UVB stimulation and decreased when the PJE was treated. As a result of the migration assay, it was confirmed that the proliferation of skin keratinocytes and the recovery rate of wounds were increased in a concentration-dependent manner. Based on the experimental results, it suggests that Petasites japonicus can be used as a functional cosmetic product that can improve skin moisturizing and skin barrier function.

• Food Science and Preservation
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• 제31권2호
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• pp.307-314
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• 2024

Broussonetia kazinoki twig extract (BKT) is recognized for its antioxidant and anti-cancer effects and natural whitening properties. So, it is used as a raw material for cosmetics. B. kazinoki twig is also an edible raw material. B. kazinoki has been used in Asia for paper production and oriental medicine, has anti-diabetic effects, and contains various flavonoids and alkaloids. In this study, to evaluate the efficacy of BKT on allergic skin inflammatory responses, we investigated its effects on factors related to skin inflammation in HaCaT keratinocytes and allergic responses in RBL-2H3 cells. There was no cytotoxicity of the 70% ethanol extract against HaCaT and RBL-2H3 cells. In HaCaT cells, stimulation with tumor necrosis factor-alpha (TNF-𝛼) and interferon-gamma (IFN-𝛾) increased the production of several chemokines, including thymus and activation-regulated chemokine (TARC), macrophage-derived chemokine (MDC), and regulated on activation, normal T cell expressed and secreted (RANTES). However, it was observed that this elevation was notably mitigated in a concentration-dependent manner upon treatment with BKT. Furthermore, BKT treatment demonstrated a significant reduction of 𝛽-hexosaminidase and inflammatory cytokines TNF-𝛼 and IL-4 in IgE-sensitized RBL-2H3 cells. Thus, it is expected that BKT can be used as a natural cosmetic and food ingredient that effectively suppresses allergic inflammatory reactions.