• Title/Summary/Keyword: Keratinocyte cells

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Downregulation of $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation in human keratinocytes by melanogenic inhibitors

  • Ahn, Kwang-Seok;Lee, Jinseon;Kim, Yeong-Shik
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.780-803
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    • 2003
  • Exposure of skin cells, particularly keratinocytes to various nuclear factor-kappaB ($\textrm{NF}_{-{\kappa}}\textrm{B}$) activators [e.g. tumor necrosis factor-$\alpha$, interleukin-1, lipopolysaccharides, and ultraviolet light] leads to phosphorylation and degradation of the inhibitory protein, $\textrm{I}_{{\kappa}}\textrm{B}$. Liberated $\textrm{NF}_{-{\kappa}}\textrm{B}$ is translocated into the nucleus where it can change or alter expression of target genes, resulting in the secretion of extracellular signaling molecules including melanotrophic factors affecting melanocyte. In order to demonstrate the possible role of $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation on the synthesis of melanotrophic factors from the keratinocytes, the activities of $\textrm{NF}_{-{\kappa}}\textrm{B}$ induced by melanogenic inhibitors (MIs) were determined in human HaCaT keratinocytes transfected with $\textrm{pNF}_{-{\kappa}}\textrm{B}$-SEAP-NPT plasmid. Transfectant cells released the secretory alkaline phosphatase (SEAP) as a transcription reporter in response to the $\textrm{NF}_{-{\kappa}}\textrm{B}$ activity and contain the neomycin phosphotransferase (NPT) gene for the dominant selection marker for geneticin resistance. MIs such as niacinamide, kojic acid, hydroquinone, resorcinol, arbutin, and glycolic acid were preincubated with transfectant HaCaT cells for 3 h and then ultraviolet B (UVB) was irradiated. $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation was measured with the SEAP reporter gene assay using a fluorescence detection method. Of the Mis tested, kojic acid ($IC_{50}$/ = 60 $\mu$M) was found to be the most potent inhibitor of UVB-upregulating $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation in transfectant HaCaT cells, which is followed by niacinamide ($IC_{50}$/= 540 $\mu$M). Pretreatment of the transfectant HaCaT cells with the Mis, especially kojic acid and niacinamide, effectively lowered $\textrm{NF}_{-{\kappa}}\textrm{B}$ binding measured by electrophoretic mobility shift assay. Furthermore, these two inhibitors remarkably reduced the secretion level of IL-6, one of melanotrophic factors, triggered by UV-radiation of the HaCaT cells. These observations suggest that Mis working at the in vivo level might act partially through the modulation of the synthesis of melanotrophic factors in keratinocyte.

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Curcumin ameliorates TNF-α-induced ICAM-1 expression and subsequent THP-1 adhesiveness via the induction of heme oxygenase-1 in the HaCaT cells

  • Youn, Gi Soo;Kwon, Dong-Joo;Ju, Sung Mi;Choi, Soo Young;Park, Jinseu
    • 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.

Cytoprotective Effect of Petasites japonicus Extract on Cadmium-induced Cytotoxicity in HaCaT cell (Cadmium으로부터 손상을 유도한 HaCaT 세포에서 머위(Petasites japonicus) 추출물의 세포보호효과)

  • Kim, Bo-Ae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.43 no.2
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    • pp.87-92
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    • 2017
  • In this study, we investigated the effect of Petasites japonicus extract on the cytotoxicity and cytoprotective effects against cadmium for cosmetics use. We measured the protein expression of apoptosis regulatory factor (Bcl-2 and procaspase-3) after treatment of Petasites japonicus extract in the cadmium-induced keratinocyte. As a result, high cell viabilities above 98% were observed in the all treated concentrations except at $200{\mu}g/mL$ of Petasites japonicus extract in keratinocytes with cadmium-induced damages. In keratinocytes with cadmium-induced damages, Bcl-2 and procaspase-3 protein expression increased in the experimental group treated with Petasites japonicus extract. Also HaCaT cells resulted in cleavage of PARP protein at 12 h post-cadmium exposure. Western blot analysis and relative density of the bands suggested that pretreatment of cells with Petasites japonicus extract inhibited cadmium-mediated cleavage of PARP. These results suggest that Petasites japonicus extract can be used as the cosmetic ingredients for cytoprotective effect.

2,3-Dimethoxy-2′-hydroxychalcone ameliorates TNF-α-induced ICAM-1 expression and subsequent monocyte adhesiveness via NF-kappaB inhibition and HO-1 induction in HaCaT cells

  • Kim, Hyejin;Youn, Gi Soo;An, Soo Yeon;Kwon, Hyeok Yil;Choi, Soo Young;Park, Jinseu
    • BMB Reports
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    • v.49 no.1
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    • pp.57-62
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    • 2016
  • Up-regulation of adhesion molecules plays an important role in the infiltration of leukocytes into the skin during the development of various inflammatory skin diseases, such as atopic dermatitis. In this study, we investigated the modulatory effects of 2,3-dimethoxy-2′-hydroxychalcone (DMHC) on tumor necrosis factor (TNF)-α-induced intercellular adhesion molecule-1 (ICAM-1) expression and monocyte adhesiveness, as well as the molecular mechanisms underlying its action in the HaCaT human keratinocyte cell line. Pre-treating HaCaT cells with DMHC significantly suppressed TNF-α-induced ICAM-1 expression and subsequent monocyte adhesiveness. DMHC inhibited TNF-α-induced activation of NF-ᴋB. In addition, DMHC induced HO-1 expression as well as NRF2 activation. Furthermore, HO-1 knockdown using siRNA reversed the inhibitory effect of DMHC on TNF-α-induced ICAM-1 expression and adhesion of monocytes to keratinocytes. These results suggest that DMHC may inhibit TNF-α-induced ICAM-1 expression and adhesion of monocytes to keratinocytes by suppressing the signaling cascades leading to NF-ᴋB activation and inducing HO-1 expression in keratinocytes. [BMB Reports 2016; 49(1): 57-62]

Anti-aging effect of Codium fragile extract on keratinocytes damaged by fine dust PM10 (미세먼지 PM10으로 손상을 유도한 각질형성세포에서 청각 (Codium fragile) 추출물의 항노화 효과)

  • Bo Ae Kim
    • The Korea Journal of Herbology
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    • v.38 no.4
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    • pp.45-52
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    • 2023
  • Objectives : Fine dust caused by environmental pollution cause oxidative damage and skin aging. In this study, The possibility of using the Codium fragile extract (CFE) as an anti-aging product for skin improvement was evaluated by confirming the protective effect of skin cells from PM10 (particulate matter 10) through inhibition of ROS and MMPs. Methods : In this study, elastase and collagenase inhibitory activities were evaluated. Cell viability was evaluated by treating keratinocytes (HaCaT cell line) with CFE at various concentrations. The cytoprotective effect from PM10 in keratinocyteswas evaluated using the 3-[4,5-dimethylthiazol]-2-yl]-2,5-diphenyl-tetrazoliumbromide (MTT) assay. ROS (reactive oxygen species) was measured in keratinocytes damaged by PM10 using DCF-DA (2′,7′-dichlorofluorescin diacetate) fluorescence staining. As an anti-aging effect of CFE, MMP-1 (matrix metalloproteinase-1) and MMP-1 (matrix metalloproteinase-9) inhibitory activities were evaluated. Results : As a result, CFE decreased the activity of elastase and collagenase. As a result of evaluating the toxicity of CFE, it is non-toxic at a concentration of 10 to 80 ㎍/㎖. Although cell viability of HaCaT cells treated with PM10 decreased, cell viability increased by 38% when treated with CFE 80 ㎍/㎖. Also, ROS decreased by 8.4%, and MMP-1 and MMP-9 decreased at CFE 80 ㎍/㎖. Conclusions : CFE showed excellent cell protection effect, and it is considered that it can be used in anti-aging products for skin improvement by effectively inhibiting ROS and MMPs from keratinocyte damage caused by fine dust.

Baicalein Protects Human Skin Cells against Ultraviolet B-Induced Oxidative Stress

  • Oh, Min Chang;Piao, Mei Jing;Jayatissa Fernando, Pattage Madushan Dilhara;Han, Xia;Madduma Hewage, Susara Ruwan Kumara;Park, Jeong Eon;Ko, Mi Sung;Jung, Uhee;Kim, In Gyu;Hyun, Jin Won
    • Biomolecules & Therapeutics
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    • v.24 no.6
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    • pp.616-622
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    • 2016
  • Baicalein (5,6,7-trihydroxy-2-phenyl-chromen-4-one) is a flavone, a type of flavonoid, originally isolated from the roots of Scutellaria baicalensis. This study evaluated the protective effects of baicalein against oxidative damage-mediated apoptosis induced by ultraviolet B (UVB) radiation in a human keratinocyte cell line (HaCaT). Baicalein absorbed light within the wavelength range of UVB. In addition, baicalein decreased the level of intracellular reactive oxygen species (ROS) in response to UVB radiation. Baicalein protected cells against UVB radiation-induced DNA breaks, 8-isoprostane generation and protein modification in HaCaT cells. Furthermore, baicalein suppressed the apoptotic cell death by UVB radiation. These findings suggest that baicalein protected HaCaT cells against UVB radiation-induced cell damage and apoptosis by absorbing UVB radiation and scavenging ROS.

TGF-β1 Expression by Proliferated Keratinocytes in the Skin of E-Irradiated Mice (E-ray를 조사한 쥐의 피부에서 증식된 keratinocyte에 의한 TGF-β1 발현)

  • Yoon, A-Ran;Kim, Do-Nyun;Seo, Min-Koo;Oh, Sang-Taek;Seo, Jung-Seon;Jun, Se-Mo;Cha, Jung-Ho;Lee, Seung-Deok;Lee, Suk-Kyeong
    • Journal of Life Science
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    • v.22 no.2
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    • pp.133-141
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    • 2012
  • In this study, we established a radiodermatitis animal model and investigated the change in immune cell proportions in the secondary lymphoid organs. The cells responsible for the increased transforming growth factor-${\beta}1$ (TGF-${\beta}1$) and interleukin-10 (IL-10) production in the lesions following irradiation were also investigated. The radiodermatitis model was constructed by locally exposing the posterior dorsal region of hairless-1 (HR-1) mice to 10 Gy electron (E)-ray/day for six consecutive days. The change in immune cell proportions was analyzed by FACS. Immunohistochemistry was carried out to detect the expression of cytokines and cell-specific markers in the skin. The proportions of antigen-presenting cells, T cells, and B cells in the lymph nodes and spleen were affected by E-irradiation. After irradiation, TGF-${\beta}1$ and IL-17 were co-localized in the papillary region of the dermis with keratin-14 (K-14)-positive cells rather than with regulatory T cells (Treg). IL-10 was not co-stained with Treg, T helper 17 (Th17) cells, dendritic cells, or macrophages. Our data indicate that TGF-${\beta}1$ is over-expressed mainly by proliferated keratinocytes in the lesions of a radiodermatitis animal model.

Preparation of Oligo Hyaluronic Acid by Hydrolysis and Its Application as a Cosmetic Ingredient (저분자량 히알루론산의 제조 및 화장품에의 응용)

  • Kim, Ki-Ho;Kim, Kyoung-Tae;Kim, Young-Heui;Kim, Jin-Guk;Han, Chang-Sung;Park, Sun-Hee;Lee, Bang-Yong
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.33 no.3
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    • pp.189-196
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    • 2007
  • In order to investigate the potential of very low molecular weight hyaluronic acid(oligo HA) as a cosmetic ingredient, we first measured its cytotoxicity in fibroblast, keratinocyte, and SIRC cell lines. For efficacy test, its moisturizing effect and penetration rate were evaluated in an artificial skin system and Caco-2 cells. Oligo HA did not show any cytotoxicity at a concentration of 300 ${\mu}g/mL$ in fibroblasts and 1,000 ${\mu}g/mL$ in keratinocytes but it showed weak proliferation. In vitro ocular test, oligo HA showed negligible cytotoxicity at the maximum concentrations used(2,000 ${\mu}g/mL$) in SIRC cells. In the test of the single and repeated cutaneous applications, oligo HA under occlusive patch did not provoke any cumulative irritation and sensitization. Oligo HA at a concentration of 0.01 % exhibited a more potent moisturizing effect than hyaluronic acid at a concentration of 0.01 %. In the permeability test using artificial skin and Caco-2 cell lines, hyaluronic acid(M.W. $1.1{\times}10^6$) was hardly observed in the down medium of the inserts. On the other hand, oligo HA(M.W. 5,000) was detected in the down medium up to 16.0 % at 6 h in Caco-2 cell culture and up to 90 % at 6 h in an artificial skin system. These results suggest that oligo HA could be useful as an active ingredient for cosmetics.

Anti-inflammatory and Antioxidant Effects of Zizyphus jujuba Miller var. inermis extracts (대추 추출물의 항산화 및 아토피 피부염 관련 항염증 효과)

  • Hong, Chang-Eui;Chun, Young-Hee;Lyu, Su-Yun
    • Korean Journal of Pharmacognosy
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    • v.50 no.2
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    • pp.86-95
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    • 2019
  • In the present study, the effect of Z. jujuba Miller var. inermis Rehder extracts on the secretion of atopic dermatitis (AD)-related cytokines and hyaluronidase activity was investigated. We prepared four fractions, butylene glycol (JB), ethanol (JE), and water (JW), with Z. jujuba Miller var. inermis extracts. JW significantly reduced the secretion of interleukin-8 and JE reduced the secretion of tumor necrosis factor-alpha in human keratinocyte HaCaT cells. Also, hyaluronidase activity was measured by enzyme assay and the fractions inhibited the activity in a dose-dependent manner. In addition, the human dermal fibroblast, HDF-n cells were treated with the extracts and antioxidant activities were measured. The results showed that the extracts increased the free radical scavenging activity and the superoxide dismutase activity. Taken together, Z. jujuba Miller var. inermis extracts reduced the secretion of AD-related cytokines and inhibited the hyaluronidase. In addition, the extracts showed antioxidant activity.

In vitro Anti-inflammation Effect of Adventitious Shoots of Toona sinesis in Propionibacterium acnes-induced Skin Dermatitis

  • Hyeon-Ji Lim;In-Sun Park;Seung-Il Jeong;Kang-Yeol Yu;Chan-Hun Jung
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.12a
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    • pp.73-73
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    • 2020
  • Toona sinensis (TS) leaf is known to antinociceptive, antioxidative stress and skin moisturizing effects. Acnes vulgaris is a chronic skin disease with various symptoms including itchiness, pain and interruption of normal skin function. Propionibacterium acnes (P. acnes) is a major factor in the occurrence of inflammatory acnes. This study evaluated the antioxidant and anti-inflammation effects by TS extract from adventitious shoots. TS extract showed anti-inflammatory activities by suppression of pro-inflammation mediators (iNOS and COX-2) in LPS-stimulated RAW264.7 cells. TS extract also has anti-inflammatory activities by inhibiting the secretion of pro-inflammatory cytokines on P. acnes-stimulated HaCaT cells. These effects were regulated by MAPK signaling pathway. Therefore, we suggest that TS extract from adventitious shoots might have applications as a medicine for treating P. acnes-induced skin diseases.

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