• Title/Summary/Keyword: HaCaT human keratinocyte cells

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Propolis Inhibits UVA-Induced Apoptosis of Human Keratinocyte HaCaT Cells by Scavenging ROS

  • Kim, Han Bit;Yoo, Byung Sun
    • Toxicological Research
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    • v.32 no.4
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    • pp.345-351
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    • 2016
  • Propolis is a resinous material collected by honeybees from several plant sources. This research aimed at showing its protective effect against UVA-induced apoptosis of human keratinocyte HaCaT cells. Using Hoechst staining, it was demonstrated that propolis (5 and $10{\mu}g/mL$) significantly inhibited the apoptosis of HaCaT cells induced by UVA-irradiation. Propolis also showed the protective effect against loss of mitochondrial membrane potential induced by UVA-irradiaiton in HaCaT cells. Propolis also inhibited the expression of activated caspase-3 induced by UVA-irradiation. To investigate the role of ROS in UVA-induced apoptosis and protection by propolis, the generation of ROS was determined in cells. The results showed that the generation of ROS was markedly reduced in cells pretreated with propolis. Consequently, propolis protected human keratinocyte HaCaT cells against UVA-induced apoptosis, which might be related to the reduction of ROS generation by UVA-irradiation.

Protectvie effects of Lonicerae Japonicae Flos against hydrogen peroxidase-induced oxidative stress on Human keratinocyte, HaCaT cells (Hydrogen peroxide로 산화적 스트레스가 유도된 HaCaT keratinocyte에서 금은화의 세포 보호 효과)

  • Seo, Seung-Hee;Choi, Mee-Ok
    • The Korea Journal of Herbology
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    • v.28 no.4
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    • pp.57-62
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    • 2013
  • 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($H_2O_2$)-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 ${\mu}M$) hydrogen peroxide. HaCaT keratinocyte cells were pre-treated with LJF (0.1, 0.25 or 0.5 mg/ml), and then stimulated with $H_2O_2$. 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 $H_2O_2$ 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 $H_2O_2$ 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 $H_2O_2$-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.

The Cytoprotective Action of Portulaca oleracea 70% EtOH Extracts via the Heme Oxygenase-1 on Hydrogen Peroxide-induced Oxidative Stress in Human Keratinocyte HaCaT Cells (마치현 70% 에탄올 추출물의 Heme Oxygenase-1 발현을 통한 산화적 스트레스에 대한 사람각질형성세포 보호 효과)

  • Seo, Seung-Hee;Jeong, Gil-Saeng
    • Korean Journal of Pharmacognosy
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    • v.46 no.2
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    • pp.116-122
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    • 2015
  • Keratinocytes are first barrier against outer challenges on skin. However, it is still largely unknown about effective protectors against ultraviolet B (UVB), and oxidative stress in human keratinocyte, HaCaT cells. Inducible heme oxygenase (HO)-1 acts against oxidants that are thought to play a role in the pathogenesis of skin disorders. Therefore, the purpose of this study was to evaluate the effect of Portulaca oleracea 70% EtOH extracts against hydrogen peroxide (H2O2)-induced oxidative stress in human keratinocytes, HaCaT cells. P. oleracea 70% EtOH extracts showed the potent protective effects on H2O2-induced toxicity by induced the expression of HO-1 in human keratinocyte, HaCaT cells. Furthermore, P. oleracea 70 % EtOH extracts caused the nuclear accumulation of nuclear factor E2-related factor 2 (Nrf2) in human keratinocytes, HaCaT cells. In addition, we found that treatment with c-Jun N-terminal kinase (JNK) inhibitor (SP600125) reduced P. oleracea 70% EtOH extracts-induced HO-1 expression, and JNK inhibitor (SP600125) also inhibited protective effects by P. oleracea 70% EtOH extracts. Therefore, these results suggest that P. oleracea 70 % EtOH extracts increases cellular resistance to H2O2-induced oxidative injury in human keratinocyte, HaCaT cells, presumably through JNK pathway-Nrf2-dependent HO-1 expression.

Investigation of the effect of Lithospermi Radix on tight-junction related genes in HaCaT cells (자초(紫草) 열수추출물이 각질형성세포 HaCaT의 세포 연접 관련 유전자의 발현에 미치는 영향 연구)

  • Cho, Namjoon;Lee, Byeongkwon;Lee, Woonghee;Kim, Keekwang;Kim, Kyoon Eon;Han, Hyosang
    • The Korea Journal of Herbology
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    • v.32 no.3
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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.

Investigation of the Effect of Sappan Lignum and Brazilin on Expression of Tight Junction Related-genes in Human Keratinocyte (소목(蘇木)과 그 지표물질인 brazilin이 인간 유래 각질 형성 세포의 tight junction 유전자 발현에 미치는 영향)

  • Cheon, Seong Hye;Choi, Sun Kyung;Cho, Nam Joon;Kim, Kee Kwang;Lee, Woong Hee;Hwang, Hyung Seo;Kim, Kyoon Eon;Han, Hyosang
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.32 no.2
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    • pp.106-112
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    • 2018
  • The aim of this research was to determine the diverse effects of Sappan Lignum extract and brazilin on human keratinocyte HaCaT cells. We confirmed the antioxidant effect of Sappan Lignum extract and brazilin was analyzed by using an ABTS assay, confirming the efficacy of water extraction method. Also, we examined effect of Sappan Lignum extract and brazilin on the cell viability, using the MTS assay in HaCaT cells. mRNA expression levels of tight junction-related genes associated with skin barrier in HaCaT cells were analyzed using quantitative real-time PCR analysis. Sappan Lignum extract increased the cellular activity of HaCaT cells and the expression of the tight junction-related genes claudin 3, claudin 6, and ZO-2. Brazilin displayed the same effects as that of the extract on HaCaT cells activity and tight junction-related genes expression. Furthermore, dispase assay demonstrated altered cell-cell adhesion strength of Sappan Lignum extract or brazilin treated HaCaT cells. Sappan Lignum extract or brazilin might be an useful ingredient in skin-mosturizinng and anti-wrinkle cosmetics, given its effects of altering mRNA expression of tight junction-related genes and enhancing cell-cell adhesion strength of HaCaT cells.

Effects of Several Seaweed Extracts on the Viability of Human Keratinocyte HaCaT Cells (각종 해조류 추출물들의 인간 피부세포주 HaCaT에 대한 활력 영향)

  • Yoon, Seung-Je;Cho, Yeon-Sook;Nam, Ju-Hyun;Lee, Hyung-Ho;Kim, Eliya;Hong, Yong-Ki
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.41 no.1
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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.

Cell proliferation and migration mechanism of caffeoylserotonin and serotonin via serotonin 2B receptor in human keratinocyte HaCaT cells

  • Kim, Hye-Eun;Cho, Hyejoung;Ishihara, Atsushi;Kim, Byungkuk;Kim, Okjoon
    • BMB Reports
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    • v.51 no.4
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    • pp.188-193
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    • 2018
  • Caffeoylserotonin (CaS), one derivative of serotonin (5-HT), is a secondary metabolite produced in pepper fruits with strong antioxidant activities. In this study, we investigated the effect of CaS on proliferation and migration of human keratinocyte HaCaT cells compared to that of 5-HT. CaS enhanced keratinocyte proliferation even under serum deficient condition. This effect of CaS was mediated by serotonin 2B receptor (5-HT2BR) related to the cell proliferation effect of 5-HT. We also confirmed that both CaS and 5-HT induced G1 progression via 5-HT2BR/ERK pathway in HaCaT cells. However, Akt pathway was additionally involved in upregulated expression levels of cyclin D1 and cyclin E induced by CaS by activating 5-HT2BR. Moreover, CaS and 5-HT induced cell migration in HaCaT cells via 5-HT2BR. However, 5-HT regulated cell migration only through ERK/AP-1/MMP9 pathway while additional Akt/NF-${\kappa}B$/MMP9 pathway was involved in the cell migration effect of CaS. These results suggest that CaS can enhance keratinocyte proliferation and migration. It might have potential as a reagent beneficial for wound closing and cell regeneration.

The effect of the cytotoxicity of sodium lauryl sulfate containing toothpaste on HaCaT and NIH-3T3 cells (구강세치제에 함유된 SLS(Sodium lauryl Sulfate)가 HaCaT 세포와 NIH-3T3 세포에 미치는 독성 효과)

  • Park, Sang-Rye;Kim, Young-Min;Choi, Byul-Bora;Kim, Ji-Young
    • Journal of Korean society of Dental Hygiene
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    • v.15 no.4
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    • pp.719-725
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    • 2015
  • Objectives: The purpose of this study was to determine the toxic effects of sodium lauryl sulfate(SLS) in human keratinocyte HaCaT cells and mouse fibroblast NIH-3T3 cells. Methods: The effect of sodium lauryl sulfate(SLS) cell viability and proliferation were determined by WST-1 assay and changes shape of nucleus were evaluated by Hoechst staining under fluorescence microscopy. Additionally, observation of cell morphological changes under light microscopy. Results: SLS induced cytotoxicity and a marked apoptosis in both HaCaT and NIH-3T3 cell lines. With the result of the WST-1 assay, SLS induced the cytotoxicity of 0.005% and 0.0075%, 0.01% SLS for 24 h after HaCaT and NIH-3T3 cells in time and dose-dependent manner(p<0.005). SLS inhibited cell growth and caused apoptosis as evidenced by nuclear fragmentation and condensation. Thus, determination of the morphological changes to define apoptosis was visualized using inverted phase contrast microscopy. Conclusions: SLS had toxicity of the human keratinocyte cells and mouse fibroblast cells and this study will provide the basic data for the development of proper SLS concentration in dentifrice.

사람 유래의 MCF10A, Chang liver및 HaCaT 세포의 소핵형성 및 세포형질전환에 미치는 2,3,7,8-Tetrachlorodibenzo-p-dioxin의 영향

  • 엄미옥;박미영;김종원;박미선;한의식;오혜영;정해관
    • Environmental Mutagens and Carcinogens
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    • v.24 no.2
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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.

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