• Journal of the Society of Cosmetic Scientists of Korea
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• v.44 no.3
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• pp.343-347
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• 2018

We synthesized 1, 2-hexanediol galactoside (HD-Gal) from HD using Escherichia coli (E. coli) ${\beta}-galactosidase$ (${\beta}-gal$), in which the reaction is generally called as transgalactosylation (reverse hydrolysis). In this study, we investigated how much HD-Gal and HD had a cytotoxic effect on HaCaT cell, in order to compare HD-Gal with HD in terms of the cytotoxicity of human skin cell. Cell proliferation assay and phase-contrast microscope observation were used for investigating the cytotoxicity. As a result, HD-Gal had not cytotoxic effect on HaCaT cell in the concentration range from 42.2 to 211 mM. In addition, when we observed the cells using microscopy, there was no change in the cell morphology. Meanwhile, when 42.2 mM and 84.4 mM HD were treated on HaCaT cell, we did not observe the cytotoxicity; however, when 168.8 mM and 211 mM HD were on HaCaT cell, HD had a higher cytotoxic effect on HaCaT cell. In addition, when HD was treated on the cells regardless of the concentration of HD, there were obvious changes in cell morphology and cell number. It was expected hopefully that HD-Gal would be applicable as a substitute for HD as a less toxic preservative in views of safety, health, and well-being.

• Journal of Life Science
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• v.29 no.10
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• pp.1164-1170
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• 2019

The present study was carried out to evaluate extracts of Ginko biloba's outer seed coat, their antioxidative effects, and their ability to protect against DNA damage due to hydrogen peroxide ($H_2O_2$) treatments in cultured human keratinocyte (HaCaT) cells. The bioassays applied for determining the antioxidant effects of a G. biloba outer seed coat water extract (GOSWE) and a G. biloba outer seed coat methanol extract (GOSME) included the DPPH and $H_2O_2$ radical scavenging assays. Our results revealed that GOSME had higher activity than GOSWE against $H_2O_2$ radical scavenging activity in in vitro and in vivo bioassays. Treatment with GOSME significantly increased the viability of $H_2O_2-treated$ HaCaT cells. GOSME's ability to protect against DNA damage was observed via the analysis of plasmids in vitro and genomic DNA in $H_2O_2-treated$ HaCaT cells. According to our data, GOSME is able to protect HaCaT cells from $H_2O_2-induced$ DNA damage and apoptosis by blocking cellular damage related to oxidative stress. In conclusion, our study indicated GOSME might serve as a novel agent for the treatment and prevention of skin disorders caused by oxidative stress.

• The Korea Journal of Herbology
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• v.34 no.1
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• pp.51-57
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• 2019

Objectives : Nypa fruticans Wurmb. (NF) have been used as a traditional medicine to treat inflammatory diseases in East-South Asia. However, it is largely undiscovered whether NF water extract could exhibit anti-inflammatory activities against tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$)-induced inflammatory responses on human keratinocytes, HaCaT cells. Therefore, this study was aimed to investigate the anti-inflammatory activity of NF water extract on TNF-${\alpha}$-induced inflammatory responses in HaCaT cells. Methods : To investigate the anti-inflammatory activites of NF water extract in HaCaT cells, the inflammatory model of HaCaT cells was established under a suitable concentration (10 ng/ml) of human TNF-${\alpha}$ (hTNF-${\alpha}$). HaCaT keratinocyte cells were pre-treated with NF water extract for 1 h, and then stimulated with hTNF-${\alpha}$. Then, the cells were harvested to measure the inflammatory mediators such as inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 and prostaglandin $E_2$ ($PGE_2$), and pro-inflammatory cytokine including TNF-${\alpha}$ and interleukin (IL)-6. In addition, we examined the inhibitory mechanisms of NF, mitogen activated protein kinases (MAPKs) and inhibitory kappa B alpha ($I{\kappa}-B{\alpha}$) Results : The treatment of NF inhibited the hTNF-${\alpha}$-induced elevation of iNOS, COX-2, and $PGE_2$ in HaCaT cells. In addition, NF treatment inhibited the hTNF-${\alpha}$-induced elevation of TNF-${\alpha}$ and IL-6. Furthermore, NF treatment inhibited the activation of MAPKs but not degradation of $I{\kappa}-B{\alpha}$. Conclusions : Taken together, our result suggest that treatment of NF could inhibit the hTNF-${\alpha}$-induced inflammatory responses via deactivation of MAPKs in HaCaT cells. This study could suggest that NF could be a beneficial agent to prevent skin damage or inflammation.

• Korean Journal of Acupuncture
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• v.39 no.2
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• pp.43-53
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• 2022

Objectives : Paeonia lactiflora Pallas (PLP) have been reported to have pharmacological effects such as anti-inflammatory and analgesic. However, it is not yet known whether PLP extract has anti-inflammatory effect on HaCaT cells, human keratinocyte. Methods : To confirm the anti-inflammatory effect of PLP on keratinocyte, TNF-𝛼/IFN-𝛾-stimulated HaCaT cells were used. HaCaT cells were pre-treated with PLP for 1h before stimulation with TNF-𝛼/IFN-𝛾. Then HaCaT cells were stimulated with TNF-𝛼/IFN-𝛾 for 24 h, the cells and media were harvested to measure the inflammatory cytokines levels. Granulocyte-macrophage colony stimulating factor (GM-CSF), monocyte chemoattractant protein-1 (MCP-1), interleukin 1 beta (IL-1𝛽), and TNF-𝛼 were analyzed by enzyme-linked immunosorbent assay (ELISA), and the mRNA expression of thymus and activation-regulated chemokines (TARC), IL-6, and IL-8 were measured by reverse transcription-polymerase chain reaction (RT-PCR). We also investigated the inhibitory mechanism of the mitogen-activated protein kinase (MAPKs) including ERK, JNK, and p38 and nuclear factor-kappaB (NF-𝜅B) by PLP using western blot. Results : PLP did not show cytotoxicity in HaCaT cells. In TNF-𝛼/IFN-𝛾-stimulated HaCaT cells, PLP significantly inhibited the expression of GM-CSF, MCP-1 IL-1𝛽, TNF-𝛼, TARC and IL-6. PLP inhibited the phosphorylation of ERK and translocation of NF-𝜅B into the nucleus. Conclusions : These results indicate that PLP could ameliorate the TNF-𝛼/IFN-𝛾-stimulated inflammatory response through inhibition of MAPK and NF-kB signal pathway. This suggests that PLP could be used beneficial agent to improve skin inflammation.

• Journal of The Korean Society of Integrative Medicine
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• v.12 no.1
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• pp.63-71
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• 2024

Purpose : Skin is the primary barrier to protect the body from various exogenous factors. Among them, UVB exposure can cause the induction of not only excessive inflammatory responses but also the degradation of extracellular matrix (ECM), including collagen and elastin. This study tried to investigate the ameliorative effect of Persicaria perfoliata ethanol extract (PPEE) on UVB-irradiated photodamage through the regulation of activator protein (AP)-1, phosphoinositide 3-kinase (PI3K)/Akt, and mitogen-activated protein kinase (MAPK) signaling molecules in HaCaT cells. Methods : The cytotoxicity of PPEE on HaCaT cells was evaluated by the WST-1 assay. The 80 mJ/cm² of UVB (312 nm) was irradiated on HaCaT cells to induce the photodamage. Western blot analysis was conducted to investigate the protein expression levels of cyclooxygenase (COX)-2, matrix metalloproteinase (MMP)-9, and heme oxygenase (HO)-1 for ameliorative status by PPEE treatment in UVB-exposed HaCaT cells. In addition, the activated status of the inflammatory transcription factor, AP-1, as well as upstream signaling molecules, PI3K/Akt, and MAPK, were also evaluated by Western blot analysis. Results : Any cytotoxic effect was not induced at the concentration up to 200 ㎍/ml by PPEE treatment. Protein expression levels of COX-2 and MMP-9 were significantly down- and up-regulated by PPEE treatment. The inflammatory transcription factor AP-1, stimulated by UVB irradiation, was also significantly attenuated by PPEE treatment. The phosphorylated status of PI3K/Akt and MAPK were mitigated by PPEE treatment in UVB-exposed HaCaT cells. Moreover, PPEE treatment potently accelerated the expression of HO-1 and its transcription factor, nuclear factor-erythroid 2-related factor (Nrf)2, which is known for its anti-inflammatory activity. Conclusion : Consequently, PPEE treatment significantly regulated COX-2 and MMP-9 expressions in UVB-irradiated HaCaT cells. The inflammatory transcription factor AP-1, along with upstream signaling molecules PI3K/Akt and MAPKs, were also attenuated by PPEE treatment in UVB-exposed HaCaT cells. Additionally, PPEE treatment exaggerated HO-1 expression and Nrf2 activation, which might have contributed to the anti-inflammatory activity of PPEE. These results indicate that PPEE could be a candidate for attenuating UVB-induced photodamage in human skin.

• Journal of Society of Preventive Korean Medicine
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• v.19 no.3
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• pp.141-154
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• 2015

Objective : In this paper, we investigated the anti-oxidative capacities and protective effects of water extract of palmul-tang (PMT) against Ultraviolet B(UVB)-induced oxidative damage in human keratinocytes(HaCaT). Method : To evaluate the anti-oxidative activities of PMT, we measured scavenging activities on 1,1-diphenyl-2-picrylhydrazyl(DPPH) radical, hydroxyl radical, hydrogen peroxide, superoxide anion, lipid peroxidation and reducing power of PMT. To give an oxidative stress to HaCaT cells, UVB was irradiated with $40mJ/cm^2$ to HaCaT cells. To detect the protective effects of PMT against UVB, we measured cell viability, apoptotic bodies and reactive oxygen species(ROS) generation. Results : PMT showed the anti-oxidative activities by scavenging DPPH radical, hydroxyl radical, hydrogen peroxide, superoxide anion, lipid peroxidation. Also PMT showed high reducing values. The UVB-induced oxidative conditions led to the cell apoptosis. However, treatment with PMT reduced oxidative stress conditions, including inhibition of cell apoptosis and expression of ROS. Conclusion : PMT had anti-oxidative activities and exhibited protective effects against UVB on HaCaT cells. PMT would be useful for the development of cosmetics treating UVB-induced skin aging.

• BMB Reports
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• v.48 no.9
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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]

• Journal of Marine Bioscience and Biotechnology
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• v.4 no.1
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• pp.31-34
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• 2010

The primary function of the skin is to protect the body from the unwanted environmental influences. The outermost layer of the skin is stratum corneum which consists of corneocytes surrounded by lipid regions. Ceramides covalently bound to keratinocytes are essential for the barrier function of the skin, which can be disturbed in the disease, like xerosis. Xerosis is an abnormal dryness of the skin which reduced the thickness of stratum corneum and ceramide content decreasing with age. In this study, 36 seaweed extracts have been tested for screening of xerosis inhibitory agent by in vitro HaCaT keratinocyte assay. Ishige sinicola and Helminthocladia australis induced the significant amount of ceramide-like substance I in HaCaT keratinocyte among the tested seaweed extracts. Sargassum fulvellum, Chondrus ecellatus and Gigartina tenella also induced the ceramide-like substance I whereas Helminthocladia australis and Pachymeniopsis elliptica induced the ceramide-like II from HaCaT keratinocyte.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.49 no.5
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• pp.589-593
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• 2016

Propionibacterium acnes infection in skin tissue often causes acne vulgaris, commonly characterized by inflammatory papules, pustules, and nodules. Chitosan and its derivatives possess strong anti-inflammatory effects. In this study, the anti-inflammatory activity of chitosan-phytochemical conjugates on P. acnes-infected human skin keratinocytes (HaCaT) was evaluated. We designed a model of P. acnes-induced inflammation in viable HaCaT cells. Nitric oxide (NO), an inflammatory marker, was successfully elevated by P. acnes infection in HaCaT cells in a dose-dependent manner. Furthermore, the levels of NO were reduced by treatment with chitosan-phytochemical conjugates (chitosan-caffeic acid, -ferulic acid and -sinapic acid) in a dose-dependent manner. Among these conjugates, chitosan-caffeic acid exhibited the strongest NO suppression in HaCaT cells infected with P. acnes. The results obtained in this study suggest that chitosan-phytochemical conjugates could be used as a potential therapeutic agent against acne vulgaris.

• 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.