• Journal of Dental Anesthesia and Pain Medicine
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• v.17 no.1
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• pp.21-28
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• 2017

Background: The skin consists of tightly connected keratinocytes, and prevents extensive water loss while simultaneously protecting against the entry of microbial pathogens. Excessive cellular levels of reactive oxygen species can induce cell apoptosis and also damage skin integrity. Propofol (2,6-diisopropylphenol) has antioxidant properties. In this study, we investigated how propofol influences intracellular autophagy and apoptotic cell death induced by oxidative stress in human keratinocytes. Method: The following groups were used for experimentation: control, cells were incubated under normoxia (5% $CO_2$, 21% $O_2$, and 74% $N_2$) without propofol; hydrogen peroxide ($H_2O_2$), cells were exposed to $H_2O_2$ ($300{\mu}M$) for 2 h; propofol preconditioning (PPC)/$H_2O_2$, cells pretreated with propofol ($100{\mu}M$) for 2 h were exposed to $H_2O_2$; and 3-methyladenine $(3-MA)/PPC/H_2O_2$, cells pretreated with 3-MA (1 mM) for 1 h and propofol were exposed to $H_2O_2$. Cell viability, apoptosis, and migration capability were evaluated. Relation to autophagy was detected by western blot analysis. Results: Cell viability decreased significantly in the $H_2O_2$ group compared to that in the control group and was improved by propofol preconditioning. Propofol preconditioning effectively decreased $H_2O_2$-induced cell apoptosis and increased cell migration. However, pretreatment with 3-MA inhibited the protective effect of propofol on cell apoptosis. Autophagy was activated in the $PPC/H_2O_2$ group compared to that in the $H_2O_2$ group as demonstrated by western blot analysis and autophagosome staining. Conclusion: The results suggest that propofol preconditioning induces an endogenous cellular protective effect in human keratinocytes against oxidative stress through the activation of signaling pathways related to autophagy.

• Journal of Microbiology and Biotechnology
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• v.18 no.7
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• pp.1308-1316
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• 2008

Propionibacterium acnes is known to playa pivotal role in the pathogenesis of acne vulgaris. CBT-SL5 is one of the antimicrobial peptides from Enterococcus faecalis SL5, and it has shown antimicrobial activity against P. acnes. The aim of this study was to investigate the anti-inflammatory effect of CBT-SL5 on the inflammation induced by P. acnes in cultured human keratinocyes. Cultured human keratinocytes derived from neonatal foreskin were treated with heat-killed P. acnes to induce inflammation, and then various concentrations of CBT-SL5 were added to the P. acnes-treated keratinocytes. The mRNA expression and protein secretion of interleukin (IL)-8, an inflammation marker, was analyzed by real-time reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. We also analyzed the nuclear factor-kappa B (NF-$\kappaB$) p65 translocation by performing immunofluorescent staining. P. acnes treatment up regulated the IL-8 mRNA expression in the keratinocytes, and this was brought about through both toll-like receptor (TLR)2 and TLR4. At the concentrations of 10, 50, and 100 ng/ml, CBT-SL5 significantly down regulated the P. acnes-induced IL-8 mRNA expression and protein production (p<0.05). At 6 hand 12 h of the treatment, CBT-SL5 significantly suppressed the P. acnes-induced IL-8 mRNA expression. Secretion of IL-8 protein was significantly reduced at 24 h. The functional inhibitory activity of CBT-SL5 was shown by CBT-SL5 suppressing the P. acnes-induced NF-$\kappaB$ translocation from the cytoplasm to the nucleus. These results demonstrated that CBT-SL5 suppressed the P. acnes-induced IL-8 expression in keratinocytes. Therefore, CBT-SL5 may be a novel anti-inflammatory treatment for acne.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.49 no.1
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• pp.59-65
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• 2023

We validated the physiological activity of Syzygium claviflorum (Roxb.) Wall. ex A.M. Cowan & Cowan (S. claviflorum) extracts (leaves, stems, fruits, and flowers) as a cosmetic ingredient. Firstly, S. claviflorum extracts removed over 80% of free radicals at various concentrations in antioxidant experiments using the DPPH and ABTS assay. In cytotoxicity experiments using human epidermal keratinocytes, S. claviflorum extracts showed low cytotoxicity. In addition, S. claviflorum extracts significantly increased the expression of keratin (KRT)1, KRT2, KRT9, KRT10, which are differentiation markers of keratinocytes, as well as genes involved in the maintenance of skin barrier function, including involucrin (IVL), loricrin (LOR), filaggrin (FLG), and claudin1 (CLDN1). In particular, the expression of FLG protein, inhibited by interleukin (IL)-4/IL-13 in atopic dermatitis, was restored by S. claviflorum extracts in in vitro experiments. Therefore, S. claviflorum extracts with excellent antioxidant efficacy and skin barrier improvement function will be useful materials for the development of future atopic dermatitis treatments and cosmetics.

• Korean Journal of Pharmacognosy
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• v.51 no.1
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• pp.49-54
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• 2020

The aim of this study was to investigate the epidermal moisturizing effects of artocarpin on normal human epidermal keratinocytes (NHEKs). To investigate the effects of artocarpin on NHEKs, cell viability and the expression of mRNAs related to skin hydration were measured. In addition, hyaluronic acid (HA)-ELISA assay was performed. Here, the effects of artocarpin on AQP3, HAS2, KRT1, and KRT10 mRNA expression, and on HA production, following UVA treatment were reported. The Quantitative real-time PCR results demonstrate that artocarpin increased AQP3, HAS2, KRT1, and KRT10 mRNA levels. The HA-ELISA assay revealed that artocarpin also increased HA production in NHEKs. Through these experiments, the epidermal moisturizing effects of artocarpin have been elucidated, providing evidence that artocarpin may be a potent cosmetic ingredient in skin anti-aging and moisturizing products. Based on these results, I anticipate that further research on the mechanisms of action of artocarpin may allow the development of not only cosmetics, but also medicines and healthcare foods.

• Food Science and Biotechnology
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• v.18 no.2
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• pp.475-480
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• 2009

Ultraviolet B (UVB) radiation provokes the generation of reactive oxygen species (ROS) in the cells and skin, which induce oxidative stress in the exposed cells, leading to photoaging and cancer. Using the human keratinocytes HaCaT cell line, we investigated the photoprotective effects of aucubin isolated from Eucommia ulmoides. Pretreatment with aucubin markedly suppressed UVB-induced oxidative stress, which manifests as a decrease in intracellular lipid peroxidation, elevation of catalase activity, and reduced glutathione content. In addition, aucubin significantly reduced expression of matrix metalloproteinase-1 (MMP-1) protein (54%) and mRNA. Taken together, these results suggest that aucubin may offer protection against UVB-induced oxidative stress and may be used as a potential agent in prevention of UVB-induced photoaging.

• Korean Journal of Pharmacognosy
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• v.52 no.4
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• pp.228-233
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• 2021

This research was carried out to investigate the moisturizing effects of Agarum cribrosum extract on normal human epidermal keratinocytes (NHEKs). Moisturizing effects of A. cribrosum extract on NHEKs were measured by quantitative realtime RT-PCR to verify the gene expressions related to skin hydration, hyaluronic acid (HA)-ELISA to detect HA production, and cell viability assays. A. cribrosum extract increased the mRNA levels of the AQP3 and HAS2 genes and HA production in NHEKs. On the other hand, A. cribrosum extract decreased the mRNA level of the KRT1 and KRT10 genes known as differentiated keratinocyte marker in NHEKs. This research showed the moisturizing effects of A. cribrosum extract. The results indicate that A. cribrosum extract can be a potent functional ingredient for skin hydration and anti-aging products. Further study is warranted regarding the use of A. cribrosum extract to develop not only cosmetics but also food and medicine.

• Natural Product Sciences
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• v.18 no.1
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• pp.52-59
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• 2012

In our continuous efforts to procure the active materials from natural products in the protective effects of oxidative stress or UV damage to skin cells we found the Prunus persica flesh extract (PPFE) is considerable to meet the demand to protect the skin damage. PPFE attenuated cell damage induced by hypoxanthine-xanthine oxidase in cultured human keratinocytes, indicating that PPFE has the potential of the scavenging effect of reactive oxygen species (ROS) in human skin cell. Moreover, PPFE significantly suppressed UVA-induced ROS production determined by the oxidation of 2,7-dichlorodihydrofluorescein diacetate (DCFH) using FACS analysis. Additional study revealed that UVA irradiation of HaCaT human keratinocytes increased the gelatinolytic activities of matrix metalloproteinase-2, and -9 (MMP-2, -9) and mRNA expression of MMP-9 analyzing by a real-time reverse transcriptase-polymerase chain reaction (RT-PCR), and these events were significantly suppressed by the treatment with PPFE. These results suggest that PPFE might be applicable as natural ingredients for skin antiaging agents via UV-induced ROS scavenging activity and suppression of MMP expression in the skin cells.

• Proceedings of the PSK Conference
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• 2001.04a
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• pp.182.2-183
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• 2001

• Journal of Biomedical Engineering Research
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• v.14 no.1
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• pp.51-62
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• 1993

An experimental study was performed for the preparation of living skin-equivalent by the using collagen gel contraction with human fibroblasts as neodermls and cultured human keratinocytes as neoderm is . The results were as follows ; 1) The rate of collagen gel contraction was dependent on the number of fibroblasts into the lattice and collagen contraction was progressed according to the increment of the number of the cells. 2) The rate of collagen gel contraction was progressed according to the decrement of the contraction of the collagen. 3) The rate of gel contraction was progressed according to the increment of serum concentration in the fixed concentration of the fibroblasts and collagen. 4) The lattice contraction was decreased according to the increment of the population doublings of the fibroblasts. 5) Macroscopically, the artificial dermis was gray white in color and tissue-like consistency and elas- ticity. 6) Microscopically, three dimensionally contracted artificial dermis showed more dense fibroblasts and its newly formed collagen fibrils in the matrix than one dimensionally contracted one. 7) Finally prepared skin-equivalent showed good attachment of living stratified keratinocytes to the dermal equivalent microscopically. It has been proposed that newly formed skin-equivalent is suitable for the graft of extensively and deeply burned patients. Shortening of the manufacturing period of skin-equivalent and development of conservation technique as a readily usable state are to be solved for our ongoing works.

• Nutrition Research and Practice
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• v.10 no.4
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• pp.371-376
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• 2016

BACKGROUND/OBJECTIVES: Chronic ultraviolet (UV) exposure-induced reactive oxygen species (ROS) are commonly involved in the pathogenesis of skin damage by activating the metalloproteinases (MMP) that break down type I collagen. Adenophora remotiflora (AR) is a perennial wild plant that inhabits Korea, China, and Japan. The present study investigated the protective effects of AR against UVB-induced photo-damage in keratinocytes. MATERIALS/METHODS: An in vitro cell-free system was used to examine the scavenging activity of 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical and nitric oxide (NO). The effect of AR on ROS formation, antioxidant enzymes, elastase, MMP-1 level, and mRNA expression of MMP-1 were determined in UVB-irradiated human keratinocyte HaCaT cells. RESULTS: AR demonstrated strong DPPH free radical and NO scavenging activity in a cell-free system exhibiting $IC_{50}$ values of 1.88 mg/mL and 6.77 mg/mL, respectively. AR pretreatment dose-dependently attenuated the production of UVB-induced intracellular ROS, and antioxidant enzymes (catalase and superoxide dismutase) were enhanced in HaCaT cells. Furthermore, pretreatment of AR prevented UVB-induced elastase and collagen degradation by inhibiting the MMP-1 protein level and mRNA expression. Accordingly, AR treatment elevated collagen content in UVB-irradiated HaCaT cells. CONCLUSION: The present study provides the first evidence of AR inhibiting UVB-induced ROS production and induction of MMP-1 as a result of augmentation of antioxidative activity in HaCaT human keratinocytes. These results suggest that AR might act as an effective inhibitor of UVB-modulated signaling pathways and might serve as a photo-protective agent.