• Title/Summary/Keyword: HaCaT Keratinocytes

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Photoprotection and Anti-inflammatory Effects of Chinese Medical Plants (약용식물추출물의 광보호 효과와 항염증 효과 연구)

  • Jin-Hwa, Kim;Sung-Min, Park;Gwan-Sub, Sim;Bum-Chun , Lee;Hyeong-Bae, Pyo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.2
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    • pp.227-233
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    • 2004
  • Chronic exposure to solar radiation, particularly ultraviolet (UV) light, causes a variety of adverse reactions on human skin, such as sunburn, photoaging and photocarcinogenesis. Free radicals and reactive oxygen species (ROS) caused by UV exposure or other environmental facts play critical roles in cellular damage. And, repeated-UV irradiation activated the expression of the matrix metalloproteinase (MMP) and induced skin irritation. Therefore, the development of effective and safe photoprotectants that can reduce and improve the skin damage has been required. The purpose of this study was to investigate the photo-protective effect of several chinese medical plants (Juniperus chinensis) on the UV -induced skin cell damages. We tested free radical and superoxide scavenging effect in vitro. Fluorometric assays of the proteolytic activities of MMP-1 (collagenase) were performed using fluorescent collagen substrates. UVA induced MMP-1 synthesis and activity were analyzed by enzyme-linked immunosorbent assay (ELISA) and gelatin-based zymography in skin fibroblasts. We also examined anti-inflammatory effects by the determination test of proinflammatory cytokine, interleukin 6 in HaCaT keratinocytes. Expression of prostaglandin E$_2$ (PGE$_2$) after UVB irradiation was measured by competitive enzyme immunoassay(EIA) using PGE$_2$ monoclonal antibody. In the human skin we tested anti-irritation effect on the SLS-induced damage skin after appling the extract containing emulsion. We found that Juniperus chinensis extract had potent radical scavenging effect by 98% at 100$\mu\textrm{g}$/mL. The extract of Juniperus chinensis showed strong inhibitory effect on MMP-1 activities by 97% at 100 $\mu\textrm{g}$/mL and suppressed the UVA induced expression of MMP-1 by 79% at 25$\mu\textrm{g}$/mL. This extract also showed strong inhibition on MMP-2 activity in UVA irradiated fibroblast by zymography. In the test of proinflammatory cytokines of human keratinocytes Juniperus chinensis extract decreased expression of interleukin 6 about 30%. The amount of PGE$_2$ by HaCaT keratinocytes was significantly increased at the doses of above 10 mJ/$\textrm{cm}^2$ of UVB (p < 0.05). At the concentrations of 3.2-25$\mu\textrm{g}$/mL of this extract, the production of PGE$_2$ by HaCaT keratinocytes (24 h after 10mJ/$\textrm{cm}^2$ UVB irradiation) was significantly inhibited in culture supernatants (p < 0.05). In SLS-induced skin irritation model in vivo, we found to reduce skin erythema and improve barrier recovery after appling Juniperus chinensis extract containing emulsion when compared to irritated non-treated and placebo-treated skin. Our results suggest that Juniperus chinensis extract can be effectively used for the prevention of UV and SLS-induced adverse skin reactions and applied as anti-aging and anti-irritation cosmetics.

UVB-Shielding Effects of para-Coumaric Acid (파라-쿠마린산의 자외선B 차단 효과)

  • Song, Kyo-Sun;Boo, Yong-Chool
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.38 no.3
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    • pp.263-273
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    • 2012
  • Recent studies have uncovered attractive properties of para-coumaric acid (PCA) as a potential skin hywhitening agent. The purpose of the current study was to examine its UVB-shielding effects. Effects of PCA on the viability of HaCaT cells exposed to UVB were assessed in vitro in comparison with other aromatic amino acid metabolites that have similar UV absorption spectra. For in vivo test, PCA cream (1.5 %) and cream base were topically applied to the dorsal skin of SKH-1 hairless mice and the inflammatory responses due to UVB exposure were monitored by changes in skin color (erythema) and thickness (edema). The cream application-UVB exposure regimen was repeated every other day for a total of 12 sessions. When HaCaT cells were irradiated with UVB, there was a dose-dependent decline in cell viability. The cell viability decline due to UVB exposure (10 mJ $cm^{-2}$) was significantly prevented by 100 ${\mu}M$ PCA, cinnamic acid, urocanic acid, or indole acrylic acid by 39, 27, 39, or 31 %, respectively. Topical application of PCA cream onto the dorsal skin of hairless mice (10 ${\mu}g\;cm^{-2}$) attenuated the changes of color parameters, $L^*$, $a^*$, $b^*$ values, and thickness of the UVB (150 mJ $cm^{-2}$)-exposed skin by 59, 50, 58, and 53 %, respectively. The current study, together with the previous studies that demonstrated the antimelanogenic effects of PCA, suggested that PCA may prevent not only dyspigmentation but also inflammatory reactions in the UVB-exposed skin.

4-O-Methylhonokiol Protects HaCaT Cells from TGF-β1-Induced Cell Cycle Arrest by Regulating Canonical and Non-Canonical Pathways of TGF-β Signaling

  • Kim, Sang-Cheol;Kang, Jung-Il;Hyun, Jin-Won;Kang, Ji-Hoon;Koh, Young-Sang;Kim, Young-Heui;Kim, Ki-Ho;Ko, Ji-Hee;Yoo, Eun-Sook;Kang, Hee-Kyoung
    • Biomolecules & Therapeutics
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    • v.25 no.4
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    • pp.417-426
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    • 2017
  • 4-O-methylhonokiol, a neolignan compound from Magnolia Officinalis, has been reported to have various biological activities including hair growth promoting effect. However, although transforming growth factor-${\beta}$ (TGF-${\beta}$) signal pathway has an essential role in the regression induction of hair growth, the effect of 4-O-methylhonokiol on the TGF-${\beta}$ signal pathway has not yet been elucidated. We thus examined the effect of 4-O-methylhonokiol on TGF-${\beta}$-induced canonical and noncanonical pathways in HaCaT human keratinocytes. When HaCaT cells were pretreated with 4-O-methylhonokiol, TGF-${\beta}1$-induced G1/G0 phase arrest and TGF-${\beta}1$-induced p21 expression were decreased. Moreover, 4-O-methylhonokiol inhibited nuclear translocation of Smad2/3, Smad4 and Sp1 in TGF-${\beta}1$-induced canonical pathway. We observed that ERK phosphorylation by TGF-${\beta}1$ was significantly attenuated by treatment with 4-O-methylhonokiol. 4-O-methylhonokiol inhibited TGF-${\beta}1$-induced reactive oxygen species (ROS) production and reduced the increase of NADPH oxidase 4 (NOX4) mRNA level in TGF-${\beta}1$-induced noncanonical pathway. These results indicate that 4-O-methylhonokiol could inhibit TGF-${\beta}1$-induced cell cycle arrest through inhibition of canonical and noncanonical pathways in human keratinocyte HaCaT cell and that 4-O-methylhonokiol might have protective action on TGF-${\beta}1$-induced cell cycle arrest.

Effects of 14 Chung-bu Medicinal Materials Described in the Dongui Bogam on Inflammatory Cytokines Production in HaCaT Keratinocytes (피부각질형성세포에서 동의보감 충부약재 14 종이 염증성 사이토카인 생성에 미치는 영향)

  • Park, Gunhyuk;Moon, Byeong Cheol;Lim, Hye-Sun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.46 no.2
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    • pp.195-204
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    • 2020
  • The purpose of this study was to investigate the effects of 14 Chung-bu medicinal materials described in the Dongui Bogam on inflammatory cytokine production in HaCaT human keratinocyte cells. In order to confirm this possibility, we screened inhibition activity of 17 cytokines using Bio-Plex ProTM Human Cytokine 17-plex assay in HaCaT cell lines. Of the 14 Chung-bu medicinal materials, Holotrichia (Ho) and Scorpio (Sc) exerted inhibitory effects on interleukin (IL)-5 production; Ho, Mantidis Ootheca (MO), and Hirudo (Hi) exerted inhibitory effects on IL-6 production; Ho, MO, Lumbricus (Lu), Hi, and Meretricis Concha (MC) showed significant inhibitory effects on IL-8 production; Gecko (Ge), Bombycis Faeces (BF), Cicadidae Periostracum (CP), and MC showed significant inhibitory effects on IL-13 production; and Testudinis Chinemis Plastrum et Carapax (TCPC), BF, and Lu exerted significant inhibitory effects on MIP-1β production. Results indicated that the Chung-bu medicinal materials might be a good candicate as potential anti-inflammatory agents for inhibition of skin inflammation. However, further investigations on these materials, including mechanistic studies, should be carried out to validated the effects in human skin equivalent models of dermatitis.

The Suppression of Inflammatory Cytokines Induced by Propionibacterium acnes Using Bacteriocin Isolated from Lactobacillus plantarum K-1 BR (Lactobacillus plantarum K-1 BR 유래 박테리오신의 여드름균에 의한 염증성 사이토카인 억제 효과)

  • Jeong, Jin Woong;Jung, Yong Hyun;Lee, Jong Sung;Yoon, Seung Won;Lee, Seung Yeon;Lee, Hong Chan;Yoon, Young Geol
    • Journal of Life Science
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    • v.26 no.8
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    • pp.970-975
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    • 2016
  • Acne vulgaris is a common chronic skin disorder that affects millions of people. The pathogenesis of acne has been known to be closely associated with the bacterium Propionibacterium acnes. Here we investigated the anti-acne activity of Lactobacillus plantarum K-1 BR by observing the expressions of proinflammatory cytokines, TNF-α, IFN-γ and IL-8, of human keratinocytes. When we applied heat-killed P. acnes to HaCaT cells, the inflammatory cytokines were induced by two- to four-fold compared to the normal control. When the bacteriocin, purified from L. plantarum K-1 BR, was pretreated to the HaCaT cells, the expression levels of TNF-α and IFN-γ stimulated by P. acnes significantly decreased to 25% and 30% of the induced levels, respectively. The IL-8 levels also significantly decreased with the concentration dependent manner of the bacteriocin. These results suggest that the bacteriocin from L. plantarum K-1 BR could reduce the expression levels of inflammatory cytokines and thus may relieve inflammations caused by acne.

Effects of Gynostemma pentaphyllum Callus Extract on Anti-inflammation, Anti-allergy, and Alleviation of Keratinocyte Damage Caused by S. aureus (돌외 캘러스 추출물의 항염, 항알러지 및 S. aureus에 의한 각질세포 손상 완화 효능)

  • Lee, Hye Suk;Lee, Hyae Min;Kim, Han Young
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.47 no.2
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    • pp.99-105
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    • 2021
  • This study was conducted to identify the efficacy related to skin barrier functions such as anti-inflammatory, anti-allergic, cornified envelope formation and the alleviation effect of keratinocyte damage caused by S. aureus using the callus extract, which was induced and extracted from the leaves of Gynostemma pentaphyllum in Ulleungdo Island. In order to confirm the anti-inflammatory effect of the G. pentaphyllum callus extract on the skin, the expression of inflammatory cytokines was investigated in primary epidermal keratinocytes (HEKa) activated with PAR-2 agonist, and the G. pentaphyllum callus extract showed IL-8, IL-25, and TSLP production inhibitory effect. β-Hexosaminidase assay using RBL-2H3 cells was performed to confirm the anti-allergic efficacy, and G. pentaphyllum callus extract showed the effect of inhibiting the release of β-hexosaminidase. In addition, G. pentaphyllum callus extract showed the cornified envelope formation effect in HaCaT cells, and through the co-culture experiment with HaCaT cells and S. aureus, it showed alleviation effect of keratinocyte damage caused by S. aureus. Therefore, G. pentaphyllum callus extract is considered to be a useful cosmetic material for improving skin barrier with anti-inflammatory, anti-allergic and alleviation effect of keratinocyte damage caused by S. aureus.

Compound K Activates Hyaluronan Synthase 2 in transformed human Keratinocytes and Fibroblasts and Increases hyaluronan in hairless mouse skin

  • Kim, Su-Jong;Kang, Byung-Yang;Cho, Si-Yang;Sung, Dae-Suk;Shin, Eiu-Suk;Chang, Hui-Kyung;Yeom, Myung-Hun;Woo, Kwang-Sik;Kim, Duk-Hee;Sim, Young-Chul;Lee, Yong-Sung
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.741-762
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    • 2003
  • Ginsenosides, the major active ingredients of ginseng, show a variety of biomedical efficacies such as anti-aging, anti oxidation and anti-inflammatory activities. To understand the effects of compound K (20-O-D-glucopyranosyl-20 (S)-protopanaxadiol), one of the major metabolite of ginsenosides on the skin, we assessed the expression level of ∼ 100 transcripts in compound K-treated HaCaT cells using cDNA microarray analysis. Compound K treatment induced differential expression of 21 genes, which have been reported to be involved in the organization of ECM structure as well as defense responses in human skin cells. One of the most interesting findings is 2-fold increase in hyaluronan synthase2 (HAS2) gene expression by compound K. We found that change in expression of HAS2 gene represents a specific response of HaCaT cells to compound K because hyaluronan synthase 1, 3 was not changed by treatment with compound K. We also demonstrated that the compound K effectively induced hyaluronan synthesis in human skin cells and hairless mouse skin. The human clinical study indicates that topical application of compound K-containing oil-in-water emulsion showed improvement of xerosis, wrinkle and fine lines in the aged skin. We concluded that compound K has anti-aging effects by the induction of HAS2 gene expression and following hyaluronan synthase.

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The Mechanism of Crinum asiaticum var. japonicum on the Activation of Anagen (문주란의 모발 성장기 유도 기전)

  • Kang, Jung-Il;Choi, Ju Hwan;Lee, Jong Gun;Yoo, Eun-Sook;Kim, Young Ho;Kang, Hee-Kyoung
    • Korean Journal of Pharmacognosy
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    • v.48 no.2
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    • pp.148-154
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    • 2017
  • Crinum asiaticum var. japonicum and its active component, norgalanthamine have been reported to have hair growth-promoting effect via the proliferation of dermal papilla cells. In this study, we investigated the other mechanisms of C. asiaticum extract var. japonicum and norgalanthamine on the hair growth. The C. asiaticum var. japonicum extract inhibited $5{\alpha}$-reductase activity by 16%, which converts testosterone to dihydrotestosterone (DHT), a main cause of androgenetic alopecia, whereas the C. asiaticum var. japonicum extract didn't function as an opener of the $K_{ATP}$ channel. On the other hand, we examined whether norgalanthamine can inhibit transforming growth factor-${\beta}$ (TGF-${\beta}$) signal pathway, which is essential in the regression induction of hair growth. Norgalanthamine inhibited the phosphorylation of Smad2/3 on TGF-${\beta}1$-induced canonical pathway in human keratinocyte HaCaT cells. These results suggested that the C. asiaticum var. japonicum extract and norgalanthamine had the potential to influence hair growth through the inhibition of $5{\alpha}$-reductase activity and TGF-${\beta}1$-induced canonical pathway.

Effect of Porcine Placenta Extract from Subcritical Water Extraction on Photodamage in Human Keratinocytes

  • Park, Yooheon;Han, Bok Kyung;Choi, Hyeon-Son;Hong, Yang Hee;Jung, Eun Young;Suh, Hyung Joo
    • Food Science of Animal Resources
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    • v.35 no.2
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    • pp.164-170
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    • 2015
  • The objective of this study was to evaluated the photoprotective effects of porcine placenta extract (PPE) on ultraviolet B (UVB)-induced oxidative stress in human keratinocytes (HaCaT) to evaluate its functional activities as a skin food ingredient. PPE prepared by subcritical water extraction was termed SPE, and subsequently digested by enzymes to prepare E-SPE. Increased intracellular reactive oxygen species (ROS) levels (192.0%) induced by UVB were decreased by SPE and E-SPE. SPE had more effective ROS scavenging activity than E-SPE treatment. UVB treatment increased expression of tissue inhibitor of metalloproteinase 1 (TIMP-1), and this elevated expression was decreased by E-SPE treatment. High-dose treatment with E-SPE (50 and 100 µg/mL) reduced TIMP-1 expression levels of UVB-C (control) to 33.5 and 34.6%, respectively. In contrast, at low SPE doses (1 and 10 µg/mL), the treatment slightly decreased TIMP- 1 expression levels to 73.3% and 71.3% of UVB-C, respectively. In conclusion, the present study demonstrated the protective effect of SPE and E-SPE against UVB damage in keratinocytes via ROS scavenging, down-regulating MMP-2 expression and up-regulating TIMP- 1 expression. This highlights the potential for SPE as an ingredient in the preparation of functional food against photoaging.

CTRP9 Regulates Growth, Differentiation, and Apoptosis in Human Keratinocytes through TGFβ1-p38-Dependent Pathway

  • Jung, Tae Woo;Park, Hyung Sub;Choi, Geum Hee;Kim, Daehwan;Lee, Taeseung
    • Molecules and Cells
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    • v.40 no.12
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    • pp.906-915
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    • 2017
  • Impairment of wound healing is a common problem in individuals with diabetes. Adiponectin, an adipocyte-derived cytokine, has many beneficial effects on metabolic disorders such as diabetes, obesity, hypertension, and dyslipidemia. C1q/TNF-Related Protein 9 (CTRP9), the closest paralog of adiponectin, has been reported to have beneficial effects on wound healing. In the current study, we demonstrate that CTRP9 regulates growth, differentiation, and apoptosis of HaCaT human keratinocytes. We found that CTRP9 augmented expression of transforming growth factor beta 1 ($TGF{\beta}1$) by transcription factor activator protein 1 (AP-1) binding activity and phosphorylation of p38 in a dose-dependent manner. Furthermore, siRNA-mediated suppression of $TGF{\beta}1$ reversed the increase in p38 phosphorylation induced by CTRP9. siRNA-mediated suppression of $TGF{\beta}1$ or p38 significantly abrogated the effects of CTRP9 on cell proliferation and differentiation while inducing apoptosis, implying that CTRP9 stimulates wound recovery through a $TGF{\beta}1$-dependent pathway in keratinocytes. Furthermore, intravenous injection of CTRP9 via tail vein suppressed mRNA expression of Ki67 and involucrin whereas it augmented $TGF{\beta}1$ mRNA expression and caspase 3 activity in skin of type 1 diabetes animal models. In conclusion, our results suggest that CTRP9 has suppressive effects on hyperkeratosis, providing a potentially effective therapeutic strategy for diabetic wounds.