• 대한화장품학회지
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• 제47권2호
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• pp.99-105
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• 2021

본 연구는 울릉도 돌외(Gynostemma pentaphyllum) 잎으로부터 캘러스 유도 및 추출한 후, 피부장벽 기능과 관련된 항염증, 항알러지, 각질세포외막 형성, S. aureus에 의한 각질형성세포 손상 완화 효능을 확인하고자 수행되었다. 돌외 캘러스 추출물의 피부에 대한 항염증 효능을 확인하기 위해, PAR-2 agonist로 활성화된 primary epidermal keratinocyte (HEKa)에서 염증성 사이토카인들의 발현을 확인한 결과, 돌외 캘러스 추출물은 IL-8, IL-25, TSLP 발현을 저해하는 효능이 있는 것을 확인하였다. RBL-2H3 세포를 이용한 β-hexosaminidase assay 시험을 통해 항알러지 효능을 확인한 결과, β-hexosaminidase 방출을 억제하는 효과를 보였다. 또한, 돌외 캘러스 추출물은 HaCaT 세포에서 각질세포외막(cornified envelope) 형성 효과가 있음을 확인하였고, HaCaT 세포와 S. aureus 공배양 실험을 통해 S. aureus에 의한 각질형성세포 생존율 감소를 완화하는 효능이 있음을 확인하였다. 본 연구 결과를 종합해보면, 울릉도 돌외 캘러스 추출물은 항염, 항알러지, S. aureus에 의한 각질세포 손상 완화 효능에 유효한 소재의 가능성을 확인하였고, 피부장벽 개선을 위한 화장품 소재로서 활용 가치가 있을 것으로 판단된다.

• 동의생리병리학회지
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• 제25권6호
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• pp.1014-1019
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• 2011

Curcumin is a natural phytochemical present in turmeric, the ground powder of the rhizomes of Curcuma longa. Curcumin has been described as having antioxidant, anti-inflammatory, and anti-carcinogenic properties. However, it is still largely unknown whether curcumin inhibits the UVB-induced inflammatory reaction in HaCaT keratinocyte cell lines. In this study, to confirm the photoprotection properties of curcumin, HaCaT keratinocyte cells were irradiated by UVB, and then treated with curcumin. UVB irradiation induced the increased expressions of IL-$1{\beta}$, TNF-${\alpha}$, IL-6, IL-8, and COX-2 in HaCaT cells. These increased expressions of cytokines (IL-$1{\beta}$, TNF-${\alpha}$, IL-6, IL-8, and COX-2) were down-regulated by curcumin treatment in UVB-irradiated HaCaT cells. In addition skin of hairless mice were damaged by UVB irradiation, which were evidenced by atrophy of epidermis and decrease of collagen in dermis. However, these damages were protected partially by co-treatment of curcumin. Taken together, this data indicate that curcumin may be a promising photoprotection agent, when used in massage pack or sunscreen product, to reduce cell death in UV-damaged skin.

• BMB Reports
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• 제56권5호
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• pp.296-301
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• 2023

Retinoic acid receptor-related orphan receptor α (RORα) plays a vital role in various physiological processes, including metabolism, cancer, circadian rhythm, cerebellar development, and inflammation. Although RORα is expressed in the skin, its role in skin physiology remains poorly elucidated. Herein, Rorα was expressed in the basal and suprabasal layers of the epidermis; however, keratinocyte-specific Rorα deletion did not impact normal epidermal formation. Under pathophysiological conditions, Rorα-deficient mice exhibited alleviated psoriasis-like symptoms, including relatively intact epidermal stratification, reduced keratinocyte hyperproliferation, and low-level expression of inflammatory cytokines in keratinocytes. Unexpectedly, the splenic population of Th17 cells was significantly lower in keratinocyte-specific RORα deficient mice than in the control. Additionally, Rorα-deficiency reduced imiquimod-induced activation of nuclear factor-κB and STAT3 in keratinocytes. Therefore, we expect that RORα inhibitors act on immune cells and keratinocytes to suppress the onset and progression of psoriasis.

• 대한본초학회지
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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.

• Biomolecules & Therapeutics
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• 제26권4호
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• pp.417-423
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• 2018

Extracellular interleukin 1 alpha (IL-$1{\alpha}$) released from keratinocytes is one of the endpoints for in vitro assessments of skin irritancy. Although cells dying via primary skin irritation undergo apoptosis as well as necrosis, IL-$1{\alpha}$ is not released in apoptotic cells. On the other hand, active secretion has been identified in interleukin-1 receptor antagonist (IL-1ra), which was discovered to be a common, upregulated, differentially-expressed gene in a microarray analysis performed with keratinocytes treated using cytotoxic doses of chemicals. This study examined whether and how IL-1ra, particularly extracellularly released IL-1ra, was involved in chemically-induced keratinocyte cytotoxicity and skin irritation. Primary cultured normal adult skin keratinocytes were treated with cytotoxic doses of chemicals (hydroquinone, retinoic acid, sodium lauryl sulfate, or urshiol) with or without recombinant IL-1ra treatment. Mouse skin was administered irritant concentrations of hydroquinone or retinoic acid. IL-1ra (mRNA and/or intracellular/extracellularly released protein) levels increased in the chemically treated cultured keratinocytes with IL-$1{\alpha}$ and IL-$1{\beta}$ mRNAs and in the chemically exposed epidermis of the mouse skin. Recombinant IL-1ra treatment significantly reduced the chemically-induced apoptotic death and intracellular/extracellularly released IL-$1{\alpha}$ and IL-$1{\beta}$ in keratinocytes. Collectively, extracellular IL-1ra released from keratinocytes could be a compensatory mechanism to reduce the chemically-induced keratinocyte apoptosis by antagonism to IL-$1{\alpha}$ and IL-$1{\beta}$, suggesting potential applications to predict skin irritation.

• International Journal of Oral Biology
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• 제46권1호
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• pp.39-44
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• 2021

This study aimed to investigate whether neurotransmitter receptors in the nervous system were also expressed in oral keratinocytes. Expressions of various neurotransmitter receptor genes in immortalized mouse oral keratinocyte (IMOK) cells were examined by reverse transcriptase polymerase chain reaction. IMOK cells expressed calcitonin gene-related peptide (CGRP) receptor subunit genes Ramp1 and Ramp3 and glutamate receptor subunit genes Grina, Gria3, Grin1, Grin2a, and Grin2d. Moreover, IMOK cells expressed Adrb2 and Chrna5 that encode beta 2 adrenergic receptor and cholinergic receptor nicotinic alpha 5 for sympathetic and parasympathetic neurotransmitters, respectively. The expression of Bdkrb1 and Ptger4, which encode receptors for bradykinin and prostaglandin E2 involved in inflammatory responses, was also observed at low levels. Expressions of Ramp1 and Grina in the mouse gingival epithelium were also confirmed by immunohistochemistry. When the function of neurotransmitter receptors expressed on IMOK cells was tested by intracellular calcium response, CGRP, glutamate, and cholinergic receptors did not respond to their agonists, but the bradykinin receptor responded to bradykinin. Collectively, oral keratinocytes express several neurotransmitter receptors, suggesting the potential regulation of oral epithelial homeostasis by the nervous system.

• Bulletin of the Korean Chemical Society
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• 제35권9호
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• pp.2787-2790
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• 2014

Quercetin is a major dietary flavonoid found in onions, apples, tea, and red wine, and potentially has beneficial effects on disease prevention. We carried out this study to investigate the effect of quercetin on UVB-induced matrix metalloproteinase-1 (MMP-1) expression in human keratinocyte HaCaT cells and to further understand the mechanisms of its action. The anti-inflammatory activity of quercetin was investigated and quercetin significantly suppressed the NO production in LPS-stimulated RAW264.7 mouse macrophages. Post treatment of quercetin decreased UV irradiation-induced phosphorylation of JNK, p38 MAPK, and ERK by 91%, 21%, and 17%, respectively. MMP-1 is mainly responsible for the degradation of dermal collagen during the aging process of human skin and quercetin suppressed the UVB-induced MMP-1 by 94%. Binding studies revealed that quercetin binds to p38 with high binding affinity ($1.85{\times}10^6M^{-1}$). The binding model showed that the 4'-hydroxy groups of the B-ring of quercetin participated in hydrogen bonding interactions with the side chains of Lys53, Glu71, and Asp168 and the 5-hydroxy group of the A-ring formed a hydrogen bond with the backbone amide of Met109. The major finding of this study shows that quercetin inhibits phosphorylation of JNK, p38 MAPK, and ERK pathway leading to the prevention of MMP-1 expression in human keratinocyte HaCaT cells. Therefore, our findings suggested the potentials of quercetin as a skin anti-photoaging agent.

• 대한의생명과학회지
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• 제8권4호
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• pp.229-234
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• 2002

Chlorella is rich in chlorella growth factor (CGF). A review of the literature has described that CGF improves the capability of a Th1-based immunity, anticancer, antioxidant antibacterial activity, growth promotion, wound healing and so on, but has not studied the effect for the metabolism and the proliferation of human skin keratinocyte. The aim of this study was to examine the effect of metabolism and the proliferation of human skin keratinocyte in vitro. CGF was extracted with an autoclaving method which is a modified hot-water extraction method from dried chlorella and conformed by means of absorbance 0.22 at 260 nm. We have measured the extracellular acidification rate (ECAR) of the CGF by Cytosensor$^{\circledR}$ Microphysiometer and evaluated responsiveness depending upon the dosage on the HaCaT cell. The ECAR for the concentrations of 0.15, 1.5, 15, 150 $\mu\textrm{g}$/ml of CGF increased as a 103.6, 128.2, 149.0 and 423.9%, respectively compared to control (0.0 $\mu\textrm{g}$/ml, 100% ECAR). The ECAR for ErbBl tyrosine kinase inhibited by 4-anilinoquinazolines, $C_{16}$H$_{14}$BrN$_3$O$_2$.HCl on tile HaCaT cells with the amounts of 10 $\mu\textrm{g}$/ml of the CCF compared with 100 $\mu\textrm{g}$/ml of rhEGF. The conclusion of the study is that CGF might increase human epidermal keratinocyte proliferation through the interaction between the epidermal growth factor receptor and itself.

• 생명과학회지
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• 제21권6호
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• pp.907-911
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• 2011

인간형 재조합단백질 각질세포 성장인자를 안정적으로 생산하는 누에 배양 세포(Bm5-hKGF cell)을 만들었다. 이 세포에서 분비되어 배지에 포함된 양은 15-20 ng/ml 정도였다. Bm5-hKGF cell에 누에의 PDI를 함께 발현시키면 세포외 분비량이 2배 증가하였다. Wound healing migration assay 결과 누에세포에서 생산된 인간형 재조합단백질 각질세포 성장인자는 세포생장을 촉진하는 활성을 가지고 있었다. 본 실험의 결과는 누에배양세포를 사용하여 저비용으로 양질의 인간형 재조합단백질을 대량생산 할 수 있는 것을 기대한다.

• 대한본초학회지
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• 제32권3호
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• pp.55-61
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• 2017

Objectives : The aim of this research was to determine the diverse effects of Lithospermi Radix Water Extract (LR) on human keratinocyte HaCaT cells, and to examine whether those effects could be applied to the human skin. Methods : We examined effect of LR on the cell viability of using the MTS assay in human keratinocyte HaCaT cells. The antioxidation effect of LR was analyzed relative to the well-known antioxidant resveratrol, using an ABTS assay. Quantitative RT-PCR analysis revealed that, in HaCaT cells, LR influenced the mRNA expression of tight-junction genes associated with skin moisturization. Furthermore, a wound-healing assay demonstrated altered cell migration in LR-treated HaCaT cells. Result : The cytotoxicity was confirmed to be higher in LR at a concentration of $800{\mu}g/m{\ell}$ using the MTS assay in HaCaT cells. In comparison to $100{\mu}M$ resveratrol, $1,600{\mu}g/m{\ell}$ LR showed either a similar or superior antioxidation effect. LR treatment in HaCaT cells reduced the mRNA expression levels of claudin 3, claudin 4, claudin 6, claudin 8, and ZO-2 to less than 0.80-fold, whereas JAM-A and Tricellulin mRNA expression level increased more than 1.33-fold. In addition, HaCaT cells migration was decreased to 83.9% by LR treatment. Conclusions : LR of antioxidation activity will have an anti-aging effect on the skin by reducing oxidative stress. Further studies are required to address the implications for human skin, given LR's effects of altering mRNA expression of tight junction-related gene and decreasing cell migration of HaCaT cells.