• 혜화의학회지
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• 제28권2호
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• pp.41-47
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• 2019

Objectives : Atopic dermatitis is an irritable skin disease accompanying rash and itching leading to impaired skin barrier. ATO-ALL is an ethanol extract of natural products comprising 12 herbs and effective on atopic dermatitis. In this study, we aimed to propose that the effect of ATO-ALL on skin regeneration in human keratinocyte cell line, HaCaT cells. Methods : To evaluate the skin regenerating effects of ATO-ALL, scratch wound healing assay, bromodeoxyuridine (BrdU) assay, and propidum iodide (PI) assay were performed using cultured HaCaT cell line. Result : Scratch wound healing assay showed that ATO-ALL was able to enhance the gap filling activity more than 2-fold at 7 ppm concentration compared with control group. BrdU assay demonstrated that ATO-ALL treatment increased the de novo cell proliferation in a dose-dependent manner. Finally, PI assay indicated that the cell cycle of HaCaT cells was modulated by ATO-ALL treatment. Conclusions : These results suggested that ATO-ALL may have skin regenerating effects by increasing cell proliferation via cell cycle regulation. Taken together, ATO-ALL is supposed to have a potential on regeneration of damaged skin or functional disease including atopic dermatitis.

• 한국환경성돌연변이발암원학회지
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• 제24권2호
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• pp.91-98
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• 2004

Although 2,3,7,8-tetrachlorodibenzo-p-dioxin(TCDD) is a powerful carcinogen in several species, limited model system exist to study carcinogenicity of this compound at cellular level. To enhance our under-standing of carcinogenicity of TCDD at cellular level, we investigated micronucleus (MN) frequency as a index of genetic toxicity and whether TCDD can transform the human cells in culture. Normal human cell lines, skin keratinocyte HaCaT, Chang liver and breast MCF10A cells were used. TCDD did not affect the cell viability of the Chang liver, HaCaT and MCF10A cells. The frequency of micronucleus was increased after treatment of TCDD for 24hr in Chang liver and HaCaT cells, but not changed in MCF10A cells. And we observed putative transformed cells in Chang liver cells exposed to 1 $\mu$M TCDD for 2 weeks. The putative transformed cells were also observed in HaCaT cells with subsequent exposure to TCDD (0.1, 1, 10, 100 nM) for 2 weeks after initial exposure to MNNG, but not observed in MCF10A cells. Collectively, these results indicate that the ability of TCDD to induce micronuclei may be involved in cellular transformation of Chang liver and HaCaT cells. Our putative TCDD-transformed cells of Chang liver and HaCaT are expected to provide a clue to the elucidation of TCDD-induced transformation pathway.

• 한국수산과학회지
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• 제41권1호
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• pp.68-72
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• 2008

The cellular viability of the human keratinocyte cell line HaCaT was compared after adding seaweed extracts to the culture medium. The viability was measured using a quick, quantitative, spectrophotometric crystal violet inclusion method. Of 36 common seaweed species tested, methanol extracts from Sargassum sagamianum and Gigartina tenella enhanced the viability of HaCaT cells by 1.6-fold, as compared to control cells, while methanol extracts from Dictyota dichotoma, Pachymeniopsis elliptica, and Enteromorpha linza decreased the viability to less than half that of controls.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 추계학술대회
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• pp.128-128
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• 2018

Nypa fruticans wurmb (NF) has been used as traditional medicinal food in Asian countries. Especially, NF has been used for conventional medicine to treat inflammatory periodontal diseases. Previous studies have been shown that NF has large amount of useful constituents such as phenolic acids, polyphenols and flavonoids. Also, NF is known as having medicinal effects such as anti-oxidant, anti-inflammatory and cholesterol-lowering effects. NF has recently been attracted to use complementary medicinal food on inflammatory diseases in Korea. However, there are no obvious effects in inflammatory and metabolic diseases also mechanisms has been studied yet. The purpose of this study was to investigate the anti-inflammatory effects of NF and steamed-NF (SNF), which recently has been used as health food, using Human keratinocyte cell line (HaCaT) and Human mast cell line (HMC-1). The cytotoxicities of NF and SNF were measured by using MTT assays in HaCaT cells and HMC-1 cell. To evaluate anti-inflammatory effects of NF and SNF, HaCaT cells were stimulated with tumor necrosis factor $(TNF)-{\alpha}$ and Interferon $(IFN)-{\gamma}$. Also, HMC-1 cells were stimulated with phorbol-12-myristate-13-acetate (PMA) and A23187 calcium ionophore (A23187) to induce allergic inflammation. Inflammatory cytokine were measured by enzyme-linked immunosorbent assay (ELISA). In this result, the extract of NF and SNF (0.01 - 1mg/ml) did not show cytotoxicity in HaCaT cells and HMC-1 cells. In addition, the NF and SNF suppressed the production of interleukin (IL)-6 and IL-8 in HaCaT cells at highest concentration. Furthermore, the treatment of SNF significantly inhibited the secretion level of IL-8 in PMA plus A23187-stimulated HMC-1 cells compared with NF treatment group. These results suggest that the extract of NF and SNF may serve as a potential therapy for skin inflammatory diseases.

• BMB Reports
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• 제48권9호
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• pp.495-500
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• 2015

Up-regulation of cell adhesion molecules and proinflammatory cytokines contributes to enhanced monocyte adhesiveness and infiltration into the skin, during the pathogenesis of various inflammatory skin diseases, including atopic dermatitis. In this study, we examined the anti-inflammatory effects of butein, a tetrahydroxychalcone, and its action mechanisms using TNF-α-stimulated keratinocytes. Butein significantly inhibited TNF-α-induced ICAM-I expression and monocyte adhesion in human keratinocyte cell line HaCaT. Butein also decreased TNF-α-induced pro-inflammatory mediators, such as IL-6, IP-10 and MCP-1, in HaCaT cells. Butein decreased TNF-α-induced ROS generation in a dose-dependent manner in HaCaT cells. In addition, treatment of HaCaT cells with butein suppressed TNF-α-induced MAPK activation. Furthermore, butein suppressed TNF-α-induced NF-kappaB activation. Overall, our results indicate that butein has immunomodulatory activities by inhibiting expression of proinflammatory mediators in keratinocytes. Therefore, butein may be used as a therapeutic agent for the treatment of inflammatory skin diseases. [BMB Reports 2015; 48(9): 495-500]

• Molecular & Cellular Toxicology
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• 제5권1호
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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.

• IMMUNE NETWORK
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• 제10권2호
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• pp.76-81
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• 2010

Ultraviolet B (UVB) induces multiple inflammatory and carcinogenic reactions. In skin, UVB induces to secrete several kinds of inflammatory cytokines from keratinocytes and also increases angiogenic process via the modulation of vascular endothelial growth factor (VEGF) production. Interleukin-21 (IL-21) is an inflammatory cytokine and produced by activated T cells. The biologic functions of IL-21 have not yet extensively studied. In the present study, we investigate the production of IL-21 from human keratinocyte cell line, HaCaT and its biological effect after exposure to UVB. First, we confirmed the IL-21 production and its receptor expression in HaCaT. And then, the change of IL-21 and VEGF production in HaCaT by UVB irradiation was examined. Not only IL-21 but also VEGF production was enhanced by UVB irradiation. Next, to determine relationship of enhanced production of IL-21 and VEGF, we detected VEGF production after neutralization of IL-21. VEGF production was reduced by IL-21 neutralization, which indicates that the IL-21 is involved in the VEGF production. Taken together, our results suggest that IL-21 and VEGF production is enhanced by UVB irradiation in HaCaT. In addition, it seems that IL-21 plays a role in the angiogenic process in skin via the modulation of VEGF production.

• 대한화장품학회지
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• 제44권3호
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• pp.343-347
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• 2018

화장품에 방부제(살균/보존제)로 사용되는 1, 2-hexanediol (HD)로 인한 부작용을 극복하기 위하여, Escherichia coli (E. coli)의 ${\beta}-galactosidase$ (${\beta}-gal$)를 이용하여 transgalactosylation 반응으로 1, 2-hexanediol galactoside (HD-Gal)를 합성하였다. 본 연구에서는 합성된 HD-Gal의 인간 피부세포에 대한 독성이 어느 정도인지를 HD와 비교하여 관찰하였다. HD-Gal과 HD의 세포독성은 인간 피부각질형성세포(HaCaT cell line)에 HD와 HD-Gal을 처리한 후, cell proliferation assay를 이용하여 비교 분석하였다. 또한 이때, 위상차 현미경으로 HD-Gal과 HD로 처리한 세포의 상태를 비교 관찰하였다. 그 결과, HD-Gal은 42.2 mM에서 211 mM의 농도 범위에서 세포독성이 관찰되지 않았으며, 현미경 관찰에서도 큰 변화를 관찰할 수 없었다. 그러나, HD의 경우에는 저농도에서(42.2 mM and 84.4 mM)는 세포독성이 관찰되지 않았으나, 고농도(168.8 and 211 mM)에서 매우 높은 세포독성을 나타내었고, 현미경 관찰에서는 고농도에서는 물론이고, 세포독성이 관찰되지 않은 HD의 저농도에서도 세포모양과 세포 수에서의 변화가 관찰되었다. 앞으로 세포독성이 감소된 HD-Gal이 HD의 대체제로서 안전, 건강 및 웰빙 개념의 새로운 용도로 개발될 수 있을 것으로 생각된다.

• 대한화장품학회지
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• 제43권4호
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• pp.329-335
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• 2017

Triterpenoid, saponin, 전분 등이 함유되어 있는 것으로 알려진 Adenophorae radix (A. radix)의 뿌리 추출물을 이용하여 각질형성세포의 분화와 피부장벽기능에 대한 연구를 수행하였다. A. radix의 뿌리 추출물은 CV-1 세포를 이용하여 $PPAR{\alpha}$ 발현을 살펴본 결과, Wy-14,643 $0.5-1.0{\mu}M$ 수준의 발현양을 나타내었다. 인체 각질형성 세포주(HaCaT)와 각질형성세포(nomal human keratinocyte)에 대한 각질형성능(cornified envelop formation, CE)은 대조군에 비해 통계적으로 유의한 증가를 보였다. HaCaT 세포에 A. radix의 뿌리 추출물 처리하였을 때, transglutaminase (TGase-1)의 유의적 증가를 보였다. A. radix의 뿌리 추출물을 함유한 간단한 화장품 제형을 약 2주간에 걸쳐 임상시험을 실시한 결과, TEWL의 유의적 감소와 수분량의 증가를 살펴볼 수 있었으며, 하박 내측에서 지질을 추출하여 세라마이드를 분석한 결과 통계적으로 유의한 증가를 관찰할 수 있었다. 이를 통하여 A. radix의 뿌리 추출물을 건조피부나 아토피 등의 피부질환과 관련된 질환의 예방 및 치료제로 사용될 수 있을 것이다.

• Biomolecules & Therapeutics
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• 제24권6호
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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.