• Title/Summary/Keyword: HaCaT cell

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Improving effect of psoriasis dermatitis by yakuchinone A in the TNF-α stimulated HaCaT cells (TNF-α 자극에 활성화된 HaCaT 세포주에서 Yakuchinone-A에 의한 건선 피부염 개선 효과)

  • Kim, Min Young;Hwang, Hyung Seo
    • Journal of Applied Biological Chemistry
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    • v.63 no.1
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    • pp.95-101
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    • 2020
  • Psoriasis is an autoimmune skin disease that is accompanied by hyper proliferation of the epidermis, erythema of various sizes, and ulceration. However, the mechanism of the development of psoriasis dermatitis is unclear. Recently, it is known that the inflammatory cytokines and Th17 cells as well as chemokine (CC motif) ligand 20 (CCL20) are involved in the process of keratinocytes hyper-differentiation, which is common in psoriasis dermatitis. Therefore, we studied the effects of yakuchinone-A, an active ingredient of Alpinia oxyphylla Miquel known for its anti-inflammatory activity, to improve psoriasis dermatitis. First, cytotoxicity of yakuchinone-A was observed in cell counting kit-8 assay and not observed in 10 ㎍/mL concentration on the human keratinocyte HaCaT cells. Yakuchinone-A in the presence of tumor necrosis factor-alpha (TNF-α) on HaCaT cells inhibited mRNA expression of IL-6, IL-8, and TNF-α by up to 61.4±7.5, 23.6±1.5, 46.0±4.8%. CCL20, a chemokine that attracts immune cells such Th17 cells to the inflammation location, was also significantly suppressed by yakuchinone-A. In addition, IκB and STAT3 phosphorylation involved in the CCL20 expression was inhibited by yakuchinone-A in a concentration-dependent manner up to the level of 79.1±5.0, 80.8±2.3%. Furthermore, yakuchinone-A downregulated CCL20 mRNA expression level on IL-17A-activated HaCaT cells as a concentration-dependent manner. Based on these results, yakuchinone-A is expected to be developed as a new material for improving psoriasis dermatitis in the future.

Profiling of Gene Expression in Human Keratinocyte Cell Line Exposed to Quantum Dot Nanoparticles

  • Kim, In-Kyoung;Lee, Seung-Ho;Kim, Yu-Ri;Seo, Sang-Hui;Jeong, Sang-Hoon;Son, Sang-Wook;Kim, Meyoung-Kon
    • Molecular & Cellular Toxicology
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    • v.5 no.1
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    • pp.51-57
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    • 2009
  • Quantum Dot (QD) nanoparticles are used in various industrial applications, such as diagnostic, drug delivery, and imaging agents of biomedicine. Although QDs are extensively used in many medical science, several studies have been demonstrated the potential toxicity of nanoparticles. The first objective of this study was to investigate the nanotoxicity of QDs in the HaCaT human keratinocyte cell line by focusing on gene expression pattern. In order to evaluate the effect of QDs on gene expression profile in HaCaT cells, we analyzed the differential genes which related to oxidative stress and antioxidant defense mechanisms by using human cDNA microarray and PCR array. A human cDNA microarray was clone set, which was sorted for a list of genes correlated with cell mechanisms. We tried to confirm results of cDNA microarray by using PCR array, which is pathway-focused gene expression profiling technology using Real-Time PCR. Although we could not find the exactly same genes in both methods, we have screened the effects of QDs on global gene expression profiles in human skin cells. In addition, our results show that QD treatment somehow regulates cellular pathways of oxidative stress and antioxidant defense mechanisms. Therefore, we suggest that this study can enlarge our knowledge of the transcriptional profile and identify new candidate biomarker genes to evaluate the toxicity of nanotoxicology.

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.

Hyaluronidase Inhibitory and Antioxidant Activities of Enzymatic Hydrolysate from Jeju Island Red Sea Cucumber (Stichopus japonicus) for Novel Anti-aging Cosmeceuticals

  • Ding, Yuling;Jiratchayamaethasakul, Chanipa;Kim, Eun-A;Kim, Junseong;Heo, Soo-Jin;Lee, Seung-Hong
    • Journal of Marine Bioscience and Biotechnology
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    • v.10 no.2
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    • pp.62-72
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    • 2018
  • An active ingredient with hyaluronidase (HAse) inhibitory effect is one of the anti-aging approaches in cosmeceuticals. Here, red sea cucumbers (RSCs), Stichopus japonicus, from Jeju Island were evaluated to examine their HAse inhibitory and antioxidant activity effect. In this study, RSCs were extracted by six enzymatic hydrolysis (Alcalase; Al, Trypsin; Try, Neutrase; Neu, Pepsin; Pep, Alpha-chymotrypsin; Chy and Protamex; Pro). Alcalase hydrolysate (AlH) showed the highest antioxidant capacities for both of oxygen radical absorbance capacity (ORAC) and trolox equivalent antioxidant capacity (TEAC) methods, compared to those of other hydrolysates, at $66.59{\pm}0.78{\mu}M\;TE/mg$ and $135.78{\pm}3.24{\mu}M\;TE/mg$, respectively. Furthermore, AlH performed the highest capacity of HAse inhibitory with $IC_{50}$ value of 3.21 mg/ml. Thus, RSCs hydrolyzed by Al were chosen to determine the cellular antioxidant activity and hyaluronic acid (HA) production effect on Human immortalized keratinocyte cell line (HaCaT). The results showed that AlH improved the cell viabilities and intracellular reactive oxygen species (ROS) induced by 2,2'-Azobis(2-amidinopropane) dihydrochloride (AAPH) were significantly decreased. In addition, AlH increased HA amount by regulating HYAL2 and HAS2 expressions in the HaCaT cells. Taken together, AlH of RSCs collected from Jeju Island showed HAse inhibitory and antioxidant activities against skin-aging which shows its potentials can be an optional natural bioactive ingredient for novel cosmeceuticals.

The Effects of Nelumbo nucifera on Utraviolet-B Irradiated human Keratinocytes (연잎, 연꽃, 연꽃 수술 추출물이 UVB 자외선 조사에 의한 각질형성세포의 보호 및 피부 노화 방지에 미치는 영향)

  • Chang, Mun-Seog;Ko, Eun-Bit;Lee, Ho-Jin;Kim, Ju-Sung;Kim, Jin-Soo;Jee, Sung-Won;Kim, Hyu-Young;Yeom, Myeong-Hoon;Kim, Duck-Hee;Kim, Han-Kon;Park, Seong-Kyu
    • The Korea Journal of Herbology
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    • v.26 no.2
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    • pp.45-49
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    • 2011
  • Objectives : The purpose of this study was to investigate the anti-aging effects on cultured human keratinocytes with Nelumbo nucifera extracts. Methods : Each parts of leaves, flowers and stamen were extracted with water or 70% ethanol. These extracts were tested for cell viability on HaCaT cells (human keratinocyte line) by MTT assay. We investigated the effects of Ultraviolet-B (UVB) irradiation on cytotoxicity and lipid peroxidation in cultured skin keratinocytes. Results : The ethanol extract of Nelumbo nucifera flowers showed maximun cell viability as 111.39% in 30 ug/ml concentration. The water extracts of stamen, flowers, leaves showed cell viability as 107.12, 101.65, 101.46%, respectively. HaCaT keratinocytes were survived 63.06% at $20mJ/cm^2$ UVB irradiation. The cell membrane lipid peroxidation was measured by accumulation malondialdehyde (MDA). The levels of MDA were decreased by the ethanol extract of Nelumbo nucifera flowers and the water extracts of stamen. Conclusions : These finding suggest that the ethanol extract of Nelumbo nucifera flowers prevent anti-aging effects on cultured human keratinocytes during UVB irradiation.

Isolation and Characterization of MMP-1 Inhibitor Peptide from Crataegus pinnatifida Bunge in Fibroblast Cell Line HS68 Cells (아가위(Crataegus pinnatifida Bunge)로 부터 HS 68세포의 MMP-1에 대한 저해활성 물질의 분리)

  • Lee, Se-Young;Chun, Hyug;Cho, Hong-Yun;An, Jeung-Hee
    • Applied Biological Chemistry
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    • v.46 no.1
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    • pp.60-65
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    • 2003
  • MMP-1 inhibitory compounds were isolated from 120 Korean traditional edible plants. UP- 1 activity significantly increased linearly with increasing UVB dose in normal human foreskin fibroblast HS68 cell, showing maximum activity at approximately 35 $mJ/cm^2$, whereas in HaCaT cell, normal human keratinocyte, no increase was observed. Maximum secretion of MMP-1 after UVB treatment occurred around 36-48 k after treatment. MMP-1 inhibitory compound isolated from cold-water fraction of Cataegus pinnatifida Bunge showed the mort potent activity. The MMP-1 inhibitory compound was deduced as a peptide based on the fact that pronase digestion decreased the activity whereas periodate oxidation did not. The most potent UP- 1-inhibitory protein, CP-2Va-2, showing an activity of 88.5% against MMP-1, was isolated through sequential column chromatography on DEAE-Toyopearl 650C, Butyl-Toyopearl 650M, and Bio-Gel P-30. Molecular weight of CP-2Va-2 determined through high performance liquid chromatography and SDS PACE was 19 and 20 kDa. respectively, signifying a monomeric structure.

Antioxidant and Tyrosinase Inhibitory Effects of the Extract Mixtures of Perilla frutescens, Houttuynia cordata and Camellia sinensis (어성초, 자소엽, 녹차 식물 추출 혼합물의 항산화 및 Tyrosinase 저해 효과에 관한 연구)

  • Lee, Kyung Eun;Lee, Eun Sun;Kang, Sang Gu
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.41 no.2
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    • pp.173-180
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    • 2015
  • In the present study, antioxidant activities and tyrosinase inhibition of Perilla frutescens, Houttuynia cordata and Camellia sinensis extracts and the extract mixtures (PHC) were investigated. PHC showed 80.2% and 98.0% of free radical scavenging activity in DPPH and ABTS analysis, respectively, and 50% tyrosinase inhibition in $1000{\mu}g/mL$ concentration. HaCaT cells did not show cell toxicity in $100{\mu}g/mL$ of the PHC. Furthermore, HaCaT cell viability by co-culture with extract H. cordata was increased more than 10% compared with untreated cells. However, the cell viability was decreased in $500{\mu}g/mL$ of the extract C. sinensis and the PHC. These results suggested that about $100{\mu}g/mL$ concentration of the PHC showed proper tyrosinase inhibitory effect and antioxidant activities. The PHC could be used as multifunctional cosmeceutical agents.

Proliferation Effect of Conditioned Medium Produced by Lymph Node Stromal Cells

  • Lee, Sang-Han;Moon, Sung-Hoon;Ha, Sang-Chul;Ma, Jin-Yeul;Kim, Min-Sook;Lee, Jin-Man;Kwon, O-Yu
    • Journal of Life Science
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    • v.12 no.1
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    • pp.22-26
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    • 2002
  • CS21 lymphoma cells that preferentially metastasize to lymph nodes after s.c. inoculation into BALB/c mice were grown in vitro in the presence of CA12 stromal cells isolated from lymph nodes. To obtain fundamental data on identification and characterization of the soluble factor(s) produced by CA12 stromal cells, we investigated the biological profile of the conditioned medium produced by CAl2 stromal cells. CAl2 conditioned medium has no affinity with Con A. CAl2 conditioned medium is associated with the proliferation of splenic T- and thymic T-cells with-out adding mitogen, although the conditioned medium cannot induce the differentiation of thymocytes. Additionally, we showed that H-7, not HA-1004 inhibits CS21 cell proliferation. These results suggest that CAl2 conditioned medium has a specific soluble factor(s) produced by lymph node stromal cells.

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Inhibition of inflammatory responses elicited by urban fine dust particles in keratinocytes and macrophages by diphlorethohydroxycarmalol isolated from a brown alga Ishige okamurae

  • Fernando, I.P. Shanura;Kim, Hyun-Soo;Sanjeewa, K.K. Asanka;Oh, Jae-Young;Jeon, You-Jin;Lee, Won Woo
    • ALGAE
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    • v.32 no.3
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    • pp.261-273
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    • 2017
  • Fine dust (FD) particles have become a major contributor to air pollution causing detrimental effects on the respiratory system and skin. Although some studies have investigated the effects of FD on the respiratory system, their possible effects on the skin remain under-explored. We investigated the FD mediated inflammatory responses in keratinocytes, present in the outer layers of skin tissues and the transfer of inflammatory potential to macrophages. We further evaluated the anti-inflammatory effects of the polyphenolic derivative, diphlorethohydroxycarmalol (DPHC) isolated from Ishige okamurae against FD-induced inflammation. Size distribution of FD particles was analyzed by scanning electron microscopy. FD particles induced the production of cyclooxygenase-2, prostaglandin E2 ($PGE_2$), interleukin (IL)-$1{\beta}$, and IL-6 in HaCaT keratinocytes and the expression of nitric oxide (NO), inducible nitric oxide synthases (iNOS), $PGE_2$, tumor necrosis factor-${\alpha}$ expression in RAW 264.7 macrophages. Further, we evaluated the inflammatory potential of the culture medium of inflammation-induced HaCaT cells in RAW 264.7 macrophages and observed a marked increase in the expression of NO, iNOS, $PGE_2$, and proinflammatory cytokines. DPHC treatment markedly attenuated the inflammatory responses, indicating its effectiveness in suppressing a broad range of inflammatory responses. It also showed anti-inflammatory potential in in-vivo experiments using FD-stimulated zebrafish embryos by decreasing NO and reactive oxygen species production, while eventing cell death caused by inflammation.

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.