인간 Keratinocyte HaCaT 세포에서 에토포사이드 또는 과산화수소에 의해 유도되는 아폽토시스에 미치는 플라보노이드들의 영향

조쌍구,안재연,강용진,최태원,이응룡 대한암예방학회 2006 Journal of cancer prevention Vol.11 No.1

Apoptosis is a fundamental cellular activity which allows for the maintenance of physiological balance and a protective mechanism against carcinogenesis. Recently, Flavonoids, a broadly distributed class of plant pigments, were known to regulate apoptosis and considerable scientific and therapeutic interest has focused on the structure and functions of these flavonoids in cancer chemotherapy. Despite of the many studies, the pro-oxidant/anti-oxidant properties of flavonoids remain debatable, and the detailed molecular mechanisms of their effects remain largely unknown. The objective, then, of the present work was to assess the apoptosis-modulating effects of several flavonoids in human keratinocyte HaCaT cells. In this study, we treated several flavonoids to human HaCaT keratinocytes and found that 3,4’-dihydroxy flavone and eriodictyol slightly increse cell viability, although other flavonoids including keamferol, quercetin, taxifolin, apigenin, naringenin and isohamnetin showed cell growth inhibition. 3,4’-dihydroxy flavone showed anti-apoptotic effect on etoposide or H2O2-induced cell death. Treatment of cells with 3,4’- dihydroxy flavone has decreased the nuclear fragmentation, PARP or pro-caspase 3 cleavage induced by etoposide or hydrogen peroxide in HaCaT keratinocytes. Taken together, our data strongly suggest that phytochemicals such as flavonoids and etoposide differentially regulate apoptosis in human HaCaT keratinocytes. (Cancer Prev Res 11, 58-64, 2006)

Hydrogen peroxide로 산화적 스트레스가 유도된 HaCaT keratinocyte에서 금은화의 세포 보호 효과

서승희 ( Seung Hee Seo ),최미옥 ( Mee Ok Choi ) 대한본초학회 2013 大韓本草學會誌 Vol.28 No.4

Objectives : Lonicerae Japonicae Flos (LJF) has been shown anti-oxidant, anti-inflammatory, anti-viral, anti-rheumatoid properties. However, it is still largely unknown whether LJF inhibits skin injury against oxidative stress in human keratinocyte, HaCaT cells. The purpose of this study was to evaluate the protective effects of LJF against hydrogen peroxide(H2O2)-induced oxidative stress in human keratinocytes, HaCaT cells. Methods : To evaluate out the protective effects of LJF on oxidative injury in HaCaT cells, an oxidative stress model of HaCaT cells was established under a suitable concentration (500 μM) hydrogen peroxide. HaCaT keratinocyte cells were pre-treated with LJF (0.1, 0.25 or 0.5 mg/ml), and then stimulated with H2O2. Then, the cells were harvested to measure the cell viability, DNA damage, and release of reactive oxygen species (ROS). Results : LJF (0.1, 0.25 or 0.5 mg/ml) itself did not show any significant toxicity in HaCaT cells. The treatment of H2O2 caused the oxidative stress, leading to the cell death, and DNA injury. However, pretreatment with LJF reduced cell death, and DNA injury. The stimulation of H2O2 on HaCaT cells resulted in excessive release of ROS, which is the main factor of oxidative stress. The excessive release of ROS was inhibited by LJF treatment significantly. Conclusions : These results could suggest that LJF exhibited the protective effects of HaCaT cells against H2O2-induced oxidative stress by inhibiting ROS release. It could be explained that LJF inhibit skin damages against oxidative stress. Thus, LJF would be useful for the development of drug or cosmetics treating skin troubles.

금은화 물추출물의 항산화 효과와 Ultraviolet(UV)B로 유도된 사람 각질형성세포 손상에 대한 보호효과

서승희 ( Seung Hee Seo ),배기상 ( Gi Sang Bae ),최선복 ( Sun Bok Choi ),조일주 ( Il Joo Jo ),김동구 ( Dong Goo Kim ),신준연 ( Joon Yeon Shin ),송호준 ( Ho Joon Song ),박성주 ( Sung Joo Park ),최미옥 ( Mee Ok Choi ) 대한본초학회 2014 大韓本草學會誌 Vol.29 No.6

Objectives : Lonicerae japonicae Flos(LJF) has been reported to exhibit anti-oxidant, anti-inflammatory, anti-viral, anti-rheumatoid properties. However, it is still largely unknown whether LJF inhibits the ultraviolet(UV)B-induced oxidative damage in human HaCaT keratinocytes. Therefore in this paper, we investigated the anti-oxidative capacity and protective effect of LJF against UVB-induced oxidative demage in human HaCaT keratinocytes. Methods : To evaluate the anti-oxidative activity of LJF extracts, we measured total phenolic contents, total flavonoid contents, antioxidant capacity, and superoxide scavenging activity. To give an oxidative stress to HaCaT cells, UVB was irradiated with 200 mJ/㎠ to HaCaT cells. To detect the protective effect of LJF against UVB, we measured cell viability, DNA fragmentation and reactive oxygen species (ROS) production. In addition, we performed high-performance liquid chromatography (HPLC) analysis to find a major component of LJF. Results : LJF contained phenolic and flavonoid contents, and showed the anti-oxidant and superoxide scavenging activity. The UVB-induced oxidative conditions led to the cell death, DNA fragmentation and reactive oxygen species (ROS) production. However, pretreatment with LJF reduced oxidative conditions, including inhibition of cell death, DNA fragmentation and ROS production. In addition, we found out chlorogenic acid as major component of LJF. Conclusions : These results could suggest that LJF contained anti-oxidative contents and exhibited protective effects against UVB on human HaCaT keratinocytes. And the effective compound of LJF which could show protective activities against UVB is chlorogenic acid. Thus, LJF and chlorogenic acid would be useful for the development of drug or cosmetics treating skin troubles.

사람의 피부상피세포에서 황색포도상구균의 독소인자인 Staphylococcal Protein A의 염증반응 촉진효과

권현진,김연정,장성희,배보경,윤화영,이희우,Kwon, Hyun-Jin,Kim, Yeon-Jung,Jang, Sung-Hee,Bae, Bo-Kyoung,Youn, Hwa-Young,Lee, Hee-Woo 한국미생물학회 2011 미생물학회지 Vol.47 No.4

Staphylococcus aureus is a major human pathogen that is associated with various types of local and systemic infection. Staphylococcal protin A (SPA), a highly expressed surface component of S. aureus, may have a role in virulence such as activating inflammation and interfering with immune clearance. We examined the effect of recombinant SPA on inflammatory response in human HaCaT keratinocytes. The recombinant SPA protein was prepared using the pET-28a Vector System in Escherichia coli. The expression of pro-inflammatory related adhesion molecules and cytokines in HaCaT cells incubated for 6, 12, and 24 h with SPA (2 ${\mu}g$/ml) was analyzed by comparative RT-PCR or ELISA. The expression of E-selectin, ICAM-1, MCP-1, IL-6 and IL-8 was significantly increased in HaCaT from 6 to 24 h after treatment with SPA. SPA showed the effect on the adhesion-promoting ability of U937 monocytes to HaCaT cells. Our data demonstrate that SPA stimulates inflammatory response of HaCaT cells, implicating an important factor for exacerbation of skin inflammation of immunologic disease. 황색포도상구균은 사람에게서 염증을 동반한 다양한 형태의 국소적 또는 전신적 감염을 일으키는 주요 병원균이며, 황색포도상구균에서 풍부하게 발현되는 Staphylococcal protein A (SPA)는 염증의 활성화나 면역 반응의 회피와 관련된 균력인자로서 작용할 수 있다. 본 연구에서는 사람의 HaCaT 피부상피세포에서 재조합 SPA 단백질을 이용하여 염증반응에 대한 효과를 조사하기 위해서 pET-28a 발현벡터시스템을 이용하여 성공적으로 재조합 SPA 단백질을 제작하였고, 이 단백질(2 ${\mu}g$/ml)을 6, 12 및 24시간 처리한 HaCaT 피부상피세포에서 RT-PCR 및 ELISA를 이용하여 염증관련 부착인자 및 사이토카인의 발현을 분석하였다. SPA 처리 후 6시간에서 24시간까지 E-selectin, ICAM-1, MCP-1, IL-6 및 IL-8의 발현이 현저하게 증가함을 확인하였다. 또한 SPA는 HaCaT 피부상피세포에 대한 U937 단핵구의 부착력을 증진시켰다. 따라서, 본 연구의 결과는 SPA가 HaCaT 피부상피세포의 염증반응을 촉진시킨다는 사실을 보여주었으며, 황색포도상구균에 의한 피부염증질환에 있어서 중요한 병원성인자로서의 역할을 수행한다는 사실을 시사해준다.

山白菊 열수추출물이 HaCaT 피부각질세포에서 미치는 항염증 효과

김세중,이규영,김근립,홍철희 대한한방안이비인후피부과학회 2022 한방안이비인후피부과학회지 Vol.35 No.2

Objectives : This study was conducted to verify the anti-inflammatory effects of Aster glehni water extracts in HaCaT keratinocytes. Methods : In this study, cell viability was confirmed by MTT assay. Production of TNF-α and IL-6 was determined by ELISA. mRNA expression of TARC and MDC were measusred by qRT-PCR. Also, expressions of p-JNK, JNK, p-ERK, ERK, p-p38, and p-38 were investigated by using western blot assay. Results : Aster glehni water extracts were not shown any significant cytotoxicity at 15.625-500㎍/㎖ in HaCaT keratinocytes. Aster glehni extracts inhibited the TNF-α and IL-6 production in HaCaT keratinocytes treated with TNF-α and IFN-γ. Also, expression of TARC, MDC, p-ERK, and p-STAT1 was decreased. Conclusions : These results suggest that Aster glehni water extracts have anti-inflammatory effects in HaCaT keratinocytes and can be applied to the development of anti-inflammatory treatment substances.

UVA 조사된 HaCaT각질형성세포에서 큰금계국 메탄올추출물의 광노화에 대한 보호 효과

오창종 ( Chang Jong Oh ),이재순 ( Jae Soon Lee ),김영철 ( Young Chul Kim ) 대한미용학회(구 대한미용과학회) 2020 대한미용학회지 Vol.16 No.4

In order to investigate the protective effects of Coreopsis lanceolate L. methanol extract (CLME) on UVA-induced photoaging in HaCaT human keratinocytes, type I procollagen production, hyaluronic acid (HA) production, and reactive oxygen species (ROS) were investigated in UVA-irradiated HaCaT cells. Treatment with 100 μg/mL CLME increased type I procollagen production by 16.6% (p<0.001), 14.0% (p<0.001), and 26.9% (p<0.001) in 10, 20, and 30 mJ/cm² UVA-irradiated HaCaT cells, respectively, which were higher than the formation of ascorbic acid (AA) (5.9%, 7.2%, and 17.6%, respectively). Similarly, treatment with 100 μg/mL CLME increased the HA production by 27.3% (p<0.001), which was higher than that of AA (24.9%, p<0.001), in 30 mJ/cm² UVA-irradiated HaCaT cells. Furthermore, treatment with 100 μg/mL CLME decreased the ROS production by 24.2% (p<0.001), which was higher than that of AA (17.2%, p<0.001), in 30 mJ/cm² UVA-irradiated HaCaT cells. The results obtained in this study showed that CLME has protective ability on UVA-induced photoaging in HaCaT cells.

P004 : Oridonin protects HaCaT keratinocytes against hydrogen peroxide-induced oxidative stress by altering microrna expression

( Hyung Jin Hahn ),( Kyu Joong Ahn ),( Sung Kwan An ),( In Sook An ),( Hwa Jun Cha ) 대한피부과학회 2013 대한피부과학회 학술발표대회집 Vol.65 No.2

Background: Oridonin, a diterpenoid isolated from Rabdosia rubescens, has been shown to exhibit anti-cancer, antimicrobial, and anti-inflammatory properties and mediate ROS production. Meanwhile, many reports demonstrate that miRNAs are key regulators in diverse biological processes in human keratinocytes. However, the relationship between ROS and miRNAs in these cells remains unclear. Objectives: We sought to demonstrate that oridonin can reduce ROS production and induce a protective effect against H2O2-induced HaCaT keratinocyte damage. Methods: Normal human HaCaT keratinocytes were treated with different doses of oridonin and a fixed dose of H2O2. Then, we used miRNA microarray and bioinformatics tools to elucidate the molecular mechanism that leads to protection against oxidative stress. Results: The data reveal that low doses of oridonin can protect against H2O2-mediated apoptosis in a concentrationand time-dependent manner. Moreover, oridonin markedly decreases H2O2-induced reactive oxygen species production in HaCaT cells. Oridonin mediates these effects by altering miRNA expression. In addition, the target genes are likely involved in cell growth and anti-apoptosis. Conclusion: In this study, we demonstrate that oridonin, a diterpenoid compound isolated from Rabdosia rubescens protects human HaCaT keratinocytes from oxidative stress induced by H2O2 exposure.

중남미 주식시장의 통합과 상호의존구조 분석

박해선,김영석 한국외국어대학교 중남미연구소 2022 중남미연구 Vol.41 No.1

This study aims to investigate the dynamic structure of interdependency of Latin American stock markets. Latin American countries try to remove all barriers to the free movement of goods, services, persons, and capital through economic integrations. In particular, the Pacific Alliance established the Latin American Integrated Market (MILA) for the free movement of capital. The vector error correction model (VECM) and directed acyclic graph(DAG) were used to investigate the dynamic structure of interdependency of stock markets among Argentina, Brazil, Chile, Colombia, Mexico, and Peru. The analysis shows that Mexico is influential to other MILA countries(Chile, Colombia, Peru) but not to Brazil and Argentina. Brazil and Argentina are influential to MILA members (Chile, Colombia, Peru) except Mexico. Peru, Colombia, and Chile have been affected by other countries. On the other hand, the stock markets of Brazil and Argentina are not interdependent. 본 연구는 중남미 주식시장의 통합과 상호의존구조를 최초로 분석했다. 중남미 국가들은 경제통합체 창설을 통해 상품, 서비스는 물론 노동과 자본의 자유로운 이동을 추진하고 있다. 특히 중남미의 대표적 경제통합체인 태평양동맹은 중남미통합증권거래소(MILA)를 창설·운영하고 있다. MILA 창설 이후 중남미 주식시장의 상호의존구조 분석 결과, 멕시코는 여타 MILA 회원국(칠레, 콜롬비아, 페루)에 영향을 미치고, 비회원국인 브라질과 아르헨티나와는 상호의존성이 낮은 것으로 나타났다. 브라질과 아르헨티나는 멕시코를 제외한 여타 MILA 회원국(칠레, 콜롬비아, 페루)에 영향을 주고 있으나, 멕시코와는 유의미한 상호의존성을 나타내지 않았다. 페루, 콜롬비아, 칠레 3개국은 여타국으로부터 일방적으로 영향을 받는 관계를 나타냈다. 한편, 브라질과 아르헨티나는 상호의존성이 없는 것으로 나타났다.

P137 Effects of alpha-mangostin on HaCaT keratinocytes differentiation

( Song Hee Han ),( Hye In Cheon ),( Min Seok Hur ),( Byung Gon Choi ),( Min Jung Kim ),( Hae Jeong Youn ),( Soo Young Kim ),( Nam Kyung Roh ),( Yang Won Lee ),( Yong Beom Choe ),( Kyu Joong Ahn ) 대한피부과학회 2016 대한피부과학회 학술발표대회집 Vol.68 No.2

<div style="display:none">fiogf49gjkf0d</div><div style="display:none">fiogf49gjkf0d</div><div style="display:none">fiogf49gjkf0d</div><div style="display:none">fiogf49gjkf0d</div><div style="display:none">fiogf49gjkf0d</div><div style="display:none">fiogf49gjkf0d</div><div style="display:none">fiogf49gjkf0d</div> Background: Alpha-mangostin(α-mangostin) is a yellow colored agent, which is the first xanthone isolated from Garcinia mangostana(mangosteen). According to previous study, α-mangostin has biological effects such as anti-inflammation and antioxidant properties. Objectives: The aim of this study was to investigate how α-mangostin promotes calcium induced differentiation of human epidermal keratinocytes, HaCaTs, and improves gloss of the nail. Methods: HaCaT cell line was cultured in DMEM media contained 10% fetal bovine serum and 1% penicillin/streptomycin, maintained by 37 ℃, 5% CO2. We measured cell viability after HaCaT keratinocyte was cultured in plate contained α-mangostin for 24hrs by using absorbance. We extracted RNA from keratinocyte and analyzed gene expression of keratin 1, involucrin. Results: Keratinocytes showed more than 85% of the cell activity and viability when exposed to less than 20 Μm α-mangostin. α-mangostin increased gene expression level of keratin1, involucrin, loricrin which are factor of differentiation of keratinocyte. It has been proven that α -mangostin is a factor to promote the differentiation of keratinocytes induced by calcium. Also α-mangostin improve nail gloss. Conclusion: α-mangostin is to facilitate the ability to differentiate induced by calcium in HaCaT keratinocytes.

Fisetin의 항산화 활성 및 미용기능식품 소재로서의 가능성 고찰

김의수,장해동,김교남 한국피부과학연구원 2012 대한피부미용학회지 Vol.10 No.3

The term of nutri-cosmetics refers to nutritional supplements which can support the function and the structure of the skin. The fisetin is a flavonol which belongs to the flavonoid group of polyphenols. Although the anti-carcinogenic property of fisetin have been widely studied, anti-oxidative function of fisetin in HaCaT keratinocytes is unknown. In this study, we examined the in vitro chemical and cellular anti-oxidant activities of fisetin in HaCaT keratinocytes and we further discussed its potential as an anti-oxidant nutricosmetics. We employed scavenging assay for the 1,1-diphenyl-2,5-picrylhydrazyl (DPPH) radicals and cellular anti-oxidant activity of fisetin was investigated in H2O2-treated HaCaT keratinocytes as judged by the 2’7’-dichlorofluorescein-diacetate (DCFH-DA)-based assay. The fisetin effectively scavenged DPPH radicals (IC50 54 μM) when compared to the scavenging activities of L-ascorbic acid (IC50 87 μM) acid and trolox (IC5074 μM), well-known anti-oxidants. In addition, the fisetin significantly ameliorates H2O2-induced oxidative stress in HaCaT keratinocytes in dose-dependent manner. The fisetin showed more strong cellular anti-oxidant activity than those of L-ascorbic acid and trolox in H2O2-treated HaCaT keratinocytes. In agreement with result of the cellular anti-oxidant activity, we observed markedly suppressed H2O2-induced oxidative stress by the pretreatment of 100 μM fisetin under the fluorescence microscopy analysis. These results provide the scientific evidences for the development of skin targeted nutri-cosmetics which has anti-oxidant function.