• Korean Journal of Plant Resources
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• v.29 no.1
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• pp.11-19
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• 2016

In this study, we investigated the anti oxidative potential and protective effects of water extract of Leonurus sibiricus L. leaf (LSLW) against ultraviolet B (UVB)-induced oxidative damage in human keratinocytes (HaCaT cells). To evaluate the anti oxidative activity of LSLW, we measured DPPH radical, hydroxyl radical, hydrogen peroxide, superoxide anion scavenging activities, lipid peroxidation inhibition, and reducing power of LSLW. For induction of oxidative stress in HaCaT cells, the cells were irradiated with UVB at 40 mJ/㎠. To investigate the protective effects of LSLW against UVB, we measured cell viability and apoptotic bodies using annexin V staining. LSLW showed anti oxidative activities by scavenging DPPH radical, hydroxyl radical, hydrogen peroxide, superoxide anion and by reducing lipid peroxidation. In addition, LSLW showed high reducing values. The UVB-induced oxidative conditions led to cell apoptosis. However, treatment with LSLW ameliorated oxidative stress conditions, including inhibition of cell death, apoptotic body. Taken together, LSLW exhibited anti oxidative and protective effects against UVB-induced damage in HaCaT cells. Thus, LSLW could be useful for the development of cosmetics for UVB-induced skin aging.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.44 no.1
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• pp.67-72
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• 2018

Green tea (Camellia sinensis L.) has been widely explored for its medicinal applications. However, most of the studies had targeted the green tea leaf, while other parts remained unexplored. In this study, protective effect of green tea root extract on Normal Human Epidermal Keratinocytes (NHEKs) against the damage induced by an external stimulant (PM2.5) was confirmed. Thirty-year-old green tea root samples were collected from Amorepacific's Dolsongi tea field and green tea root extract was prepared with 70% ethanol. Total crude saponin content in green tea root extract was 54%, which is much higher than that in ginseng extract. Our results suggest that green tea root extract can be used as a natural surfactant in cosmetics. For evaluating its protective effect against the damage induced by PM2.5, IL-36G was used as a biomarker. IL-36G mRNA expression level increased remarkable upon PM2.5 treatment in NHEKs. Moreover, IL-36G was recently reported to be expressed in psoriasis lesions. Results showed significant decrease of IL-36G expression by treatment of green tea root extract. In conclusion, thirty-year-old green tea root extract can be used as a natural surfactant with a high saponin content and may have protective effect against the damage induced by PM2.5.

• International Journal of Oral Biology
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• v.39 no.2
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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.

• Korean Journal of Microbiology
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• v.47 no.4
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• pp.348-355
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• 2011

Staphylococcus aureus is a major human pathogen that is associated with various types of local and systemic infection. Staphylococcal protin A (SPA), a highly expressed surface component of S. aureus, may have a role in virulence such as activating inflammation and interfering with immune clearance. We examined the effect of recombinant SPA on inflammatory response in human HaCaT keratinocytes. The recombinant SPA protein was prepared using the pET-28a Vector System in Escherichia coli. The expression of pro-inflammatory related adhesion molecules and cytokines in HaCaT cells incubated for 6, 12, and 24 h with SPA (2 ${\mu}g$/ml) was analyzed by comparative RT-PCR or ELISA. The expression of E-selectin, ICAM-1, MCP-1, IL-6 and IL-8 was significantly increased in HaCaT from 6 to 24 h after treatment with SPA. SPA showed the effect on the adhesion-promoting ability of U937 monocytes to HaCaT cells. Our data demonstrate that SPA stimulates inflammatory response of HaCaT cells, implicating an important factor for exacerbation of skin inflammation of immunologic disease.

• Korean Journal of Food Science and Technology
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• v.50 no.6
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• pp.653-659
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• 2018

In this study, we investigated the antioxidant activities and anti-aging efficacy in terms of suppression of matrix metalloproteinases (MMPs) in UVB-irradiated HaCaT cells by adding the ethanol extracts of Josaengheogchal (JE) and Shintoheug rice (SRE). In the electron-donating ability and ABTS radical-scavenging assays, we observed that both JE and SRE had scavenging activities and in a collagenase inhibition assay, both extracts showed inhibition effects of over 73% at $1,000{\mu}g/mL$ concentration. The expression of MMP-1 and -3, when the extracts were treated with UVB $50mJ/cm^2$, irradiated human HaCaT keratinocytes, was analyzed by western blotting and reverse-transcription polymerase chain reaction (RT-PCR). The results showed that MMP-1 and -3 proteins and mRNAs were downregulated in a concentration-dependent manner in response to both extracts. Therefore, we expect that these compounds have a potential for the use as functional ingredients with anti-aging effects in the cosmetic and food industries.

• Journal of Periodontal and Implant Science
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• v.49 no.5
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• pp.270-286
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• 2019

Purpose: Despite the well-known anti-inflammatory effects of vitamin D in periodontal health, its mechanism has not been fully elucidated. In the present study, the effect of vitamin D on strengthening E-cadherin junctions (ECJs) was explored in human gingival keratinocytes (HGKs). ECJs are the major type of intercellular junction within the junctional epithelium, where loose intercellular junctions develop and microbial invasion primarily occurs. Methods: HOK-16B cells, an immortalized normal human gingival cell line, were used for the study. To mimic the inflammatory environment, cells were treated with tumor necrosis factor-alpha ($TNF-{\alpha}$). Matrix metalloproteinases (MMPs) in the culture medium were assessed by an MMP antibody microarray and gelatin zymography. The expression of various molecules was investigated using western blotting. The extent of ECJ development was evaluated by comparing the average relative extent of the ECJs around the periphery of each cell after immunocytochemical E-cadherin staining. Vitamin D receptor (VDR) expression was examined via immunohistochemical analysis. Results: $TNF-{\alpha}$ downregulated the development of the ECJs of the HGKs. Dissociation of the ECJs by $TNF-{\alpha}$ was accompanied by the upregulation of MMP-9 production and suppressed by a specific MMP-9 inhibitor, Bay 11-7082. Exogenous MMP-9 decreased the development of ECJs. Vitamin D reduced the production of MMP-9 and attenuated the breakdown of ECJs in the HGKs treated with $TNF-{\alpha}$. In addition, vitamin D downregulated $TNF-{\alpha}$-induced nuclear factor kappa B ($NF-{\kappa}B$) signaling in the HGKs. VDR was expressed in the gingival epithelium, including the junctional epithelium. Conclusions: These results suggest that vitamin D may avert $TNF-{\alpha}$-induced downregulation of the development of ECJs in HGKs by decreasing the production of MMP-9, which was upregulated by $TNF-{\alpha}$. Vitamin D may reinforce ECJs by downregulating $NF-{\kappa}B$ signaling, which is upregulated by $TNF-{\alpha}$. Strengthening the epithelial barrier may be a way for vitamin D to protect the periodontium from bacterial invasion.

• Journal of Applied Biological Chemistry
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• v.65 no.4
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• pp.405-412
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• 2022

Echinodorus cordifolius (L.) is an aquatic plant in the family Alismataceae. The anti-skin aging activity of E. cordifolius (L.) has not been yet reported. Therefore, the objective of the present study was to prepare 70% ethanol extract (ECEE) from E. cordifolius (L.) and investigate their antioxidant and anti-hyaluronidase activities for confirm the potential of anti-skin aging. ECEE showed good activities of DPPH, hydrogen peroxide scavenging, and hyaluronidase inhibition, with EC₅₀ and IC₅₀ values of 31.4, 300, and 450 ㎍/mL, respectively. ECEE also significantly improved cell viability and inhibited intracellular reactive oxygen species dose-dependently against 1 mM hydrogen peroxide-induced oxidative stress in immortalized human keratinocytes (HaCaT cells). Furthermore, ECEE upregulated hyaluronic acid (HA)-synthesizing enzyme hyaluronan synthase 2 (HAS2) expression level, but downregulated expression level of HA-degrading enzyme hyaluronidase 2, resulting in increased HA production in HaCaT cells. Taken together, these results suggest that ECEE shows antioxidant and anti-hyaluronidase potential and could be a functional cosmetic ingredients for anti-skin aging.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.356-358
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• 2003

The use of cultured epidermal sheet grafts for large full-thickness wound has been tempered by weak points such as long culture periods, difficulty in handling the fragile sheets, high costs and the detachment of the skin cells from the culture dishes by enzymatic digestion. To overcome the drawback of epidermal sheet grafts, we have developed a transplantation method to spray the cultured human keratinocytes with the mixtures of rhEGF and fibrin gel matrix to the full-thickness wounds on the dorsum of the athymic mice to regenerate epithelial layers. Wound biopsies were retrieved at 7, 14 and 21 days after transplantation and retrieved biopsies were analyzed by histology and immunohistochemistry. Transplanted keratinocytes and EGF-fibriin gel accelerated wound regeneration compared with control groups. The technique developed in this study overcomes the drawbacks of the current cultured epidermal sheet grafts and accelerates epidermal wound healing.

• Biomolecules & Therapeutics
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• v.23 no.6
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• pp.525-530
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• 2015

Ceramide is the most abundant lipid in the epidermis and plays a critical role in maintaining epidermal barrier function. Overall ceramide content in keratinocyte increases in parallel with differentiation, which is initiated by supplementation of calcium and/or vitamin C. However, the role of metabolic enzymes responsible for ceramide generation in response to vitamin C is still unclear. Here, we investigated whether vitamin C alters epidermal ceramide content by regulating the expression and/or activity of its metabolic enzymes. When human keratinocytes were grown in 1.2 mM calcium with vitamin C ($50{\mu}g/ml$) for 11 days, bulk ceramide content significantly increased in conjunction with terminal differentiation of keratinocytes as compared to vehicle controls (1.2 mM calcium alone). Synthesis of the ceramide fractions was enhanced by increased de novo ceramide synthesis pathway via serine palmitoyltransferase and ceramide synthase activations. Moreover, sphingosine-1-phosphate (S1P) hydrolysis pathway by action of S1P phosphatase was also stimulated by vitamin C supplementation, contributing, in part, to enhanced ceramide production. However, activity of sphingomyelinase, a hydrolase enzyme that converts sphingomyelin to ceramide, remained unaltered. Taken together, we demonstrate that vitamin C stimulates ceramide production in keratinocytes by modulating ceramide metabolicrelated enzymes, and as a result, could improve overall epidermal barrier function.

• BMB Reports
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• v.46 no.8
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• pp.410-415
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• 2013

Adhesion molecules such as ICAM-1 are important in the infiltration of leukocytes into the site of inflammation. In this study, we investigated the inhibitory effects of curcumin on ICAM-1 expression and monocyte adhesiveness as well as its underlying action mechanism in the TNF-${\alpha}$-stimulated keratinocytes. Curcumin induced expression of heme oxygenase-1 (HO-1) in the human keratinocyte cell line HaCaT. In addition, curcumin induced Nrf2 activation in dose- and time-dependent manners in the HaCaT cells. Curcumin suppressed TNF-${\alpha}$-induced ICAM-1 expression and subsequent monocyte adhesion, which were reversed by the addition of tin protoporphyrin IX (SnPP), a specific inhibitor of HO-1, or HO-1 knockdown using siRNA. Furthermore, Nrf2 knockdown using siRNA reversed the inhibitory effect of curcumin on the TNF-${\alpha}$-induced ICAM-1 expression and adhesion of monocytes to keratinocytes. These results suggest that curcumin may exert its anti-inflammatory activity by suppressing the TNF-${\alpha}$-induced ICAM-1 expression and subsequent monocyte adhesion via expression of HO-1 in the keratinocytes.