• The Journal of Korean Medicine
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• v.43 no.3
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• pp.1-15
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• 2022

Objectives: Myrrh have been used as a traditional remedy to treat infectious and inflammatory diseases. However, it is largely unknown whether myrrh ethanol extract could exhibit the inhibitory activities against particulate matter (PM)-induced skin injury on human keratinocytes, HaCaT cells. Therefore, this study was aimed to investigate the inhibitory activity of myrrh ethanol extract on PM-induced skin injury in HaCaT cells. Methods: To investigate the inhibitory effects of myrrh ethanol extract in HaCaT cells, the skin injury model of HaCaT cells was established under PM treatment. HaCaT keratinocyte cells were pre-treated with myrrh ethanol extract for 1 h, and then stimulated with PM. Then, the cells were harvested to measure the cell viability, reactive oxygen species (ROS), pro-inflammatory cytokines including interleukin (IL) 1-beta, IL-6, and tumor necrosis factor (TNF)-𝛼, hyaluronidase, collagen, MMPs. In addition, we examined the mitogen activated protein kinases (MAPKs) and inhibitory kappa B alpha (I𝜅-B𝛼) as inhibitory mechanisms of myrrh ethanol extract. Results: The treatment of myrrh ethanol extract inhibited the PM-induced cell death and ROS production in HaCaT cells. In addition, myrrh ethanol extract treatment inhibited the PM-induced elevation of IL-1beta, IL-6, and TNF-𝛼. Also, myrrh ethanol extract treatment inhibited the increase of hyaluronidase, MMP and decrease of collagen. Furthermore, myrrh ethanol extract treatment inhibited the activation of MAPKs and the degradation of I𝜅-B𝛼. Conclusions: Our result suggest that treatment of myrrh ethanol extract could inhibit the PM-induced skin injury via deactivation of MAPKs and nuclear factor (NF)-𝜅B in HaCaT cells. This study could suggest that myrrh ethanol extract could be a beneficial agent to prevent skin damage or inflammation.

• Korean Journal of Agricultural Science
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• v.48 no.4
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• pp.1003-1013
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• 2021

Thioredoxin (TRX) protein is an antioxidant responsible for reducing other proteins by exchanging cysteine thiol-disulfide and is also known for its anti-allergic and anti-aging properties. On the other hand, epidermal growth factor (EGF) is an important material used in the cosmetics industry and an essential protein necessary for dermal wound healing facilitated by the proliferation and migration of keratinocytes. EGF also assists in the formation of granulation tissues and stimulates the motility of fibroblasts. Hence, genetically modified soybeans were developed to overexpress these industrially important proteins for mass production. A single-dose oral-toxicity-based study was conducted to evaluate the potential toxic effects of TRX and EGF proteins, as safety assessments are necessary for the commercial use of seed-specific protein-expressing transgenic soybeans. To achieve this rationale, TRX and EGF proteins were mass purified from recombinant E. coli. The single-dose oral-toxicity tests of the TRX and EGF proteins were carried out in six-week old male and female Institute of Cancer Research (ICR) mice. The initial evaluation of the single-dose TRF and EGF treatments was based on monitoring the toxicity signatures and mortality rates among the mice, and the resultant mortality rates did not show any specific clinical symptoms related to the proteins. Furthermore, no significant differences were observed in the weights between the treatment and control groups of male and female ICR mice. After 14 days of treatment, no differences were observed in the autopsy reports between the various treatment and control groups. These results suggest that the minimum lethal dose of TRX and EGF proteins is higher than the allowed 2,000 mg·kg^-1 limit.

• Biomolecules & Therapeutics
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• v.31 no.6
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• pp.640-647
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• 2023

The skin, the largest organ in the body, undergoes age-related changes influenced by both intrinsic and extrinsic factors. The primary external factor is photoaging which causes hyperpigmentation, uneven skin surface, deep wrinkles, and markedly enlarged capillaries. In the human dermis, it decreases fibroblast function, resulting in a lack of collagen structure and also decreases keratinocyte function, which compromises the strength of the protective barrier. In this study, we found that treatment with γ-aminobutyric acid (GABA) had no toxicity to skin fibroblasts and GABA enhanced their migration ability, which can accelerate skin wound healing. UVB radiation was found to significantly induce the production of matrix metalloproteinase 1 (MMP-1), but treatment with GABA resulted in the inhibition of MMP-1 production. We also investigated the enhancement of filaggrin and aquaporin 3 in keratinocytes after treatment with GABA, showing that GABA can effectively improve skin moisturization. In vivo experiments showed that oral administration of GABA significantly improved skin wrinkles and epidermal thickness. After the intake of GABA, there was a significant decrease observed in the increase of skin thickness measured by calipers and erythema. Additionally, the decrease in skin moisture and elasticity in hairless mice exposed to UVB radiation was also significantly restored. Overall, this study demonstrates the potential of GABA as functional food material for improving skin aging and moisturizing.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2021.04a
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• pp.49-49
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• 2021

The roots of reed (Phragmites australis) were used in traditional medicine to treat respiratory problems, including symptoms such as high fever and cough. In this study, we identified the active ingredient from 70% EtOH reed root extract, and evaluated the whitening, wrinkle improvement and moisturizing effects. The content of p-coumaric acid, the active ingredient of the roots of P. australis, was slightly lower in 70% EtOH extract than in 100% EtOH extract. However, 70% EtOH reed root extract showed similar or higher effect in reducing power, DPPH, hydrogen peroxide scavenging, and nitric oxide scavenging activity compared to 100% EtOH extract. Moreover, 70% EtOH reed root extract markedly inhibited melanogenesis in B16F10 cells treated with α-melanocyte-stimulating hormone. 70% EtOH reed root extract significantly inhibited the mRNA expression of matrix metalloproteinase-1 (MMP-1) and reduced elastase activity in HDF human dermal fibroblasts. In addition, 70% EtOH reed root extract ameliorated hyaluronic acid synthase-2 (HAS-2) expression induced by ultraviolet B (UVB) stimulation in HaCaT keratinocytes. The results of this study suggest that 70% EtOH reed root extract has potential as a functional cosmetic material related to whitening, wrinkle improvement, and moisturizing.

• Biomolecules & Therapeutics
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• v.32 no.4
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• pp.499-507
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• 2024

Specific sensitivity of the skin to ultraviolet B (UVB) rays is one of the mechanisms responsible for widespread skin damage. This study tested whether 1,3,5-trihydroxybenzene (THB), a compound abundant in marine products, might inhibit UVB radiationinduced NADPH oxidase 4 (NOX4) in both human HaCaT keratinocytes and mouse dorsal skin and explore its cytoprotective mechanism. The mechanism of action was determined using western blotting, immunocytochemistry, NADP⁺/NADPH assay, reactive oxygen species (ROS) detection, and cell viability assay. THB attenuated UVB-induced NOX4 expression both in vitro and in vivo, and suppressed UVB-induced ROS generation via NADP⁺ production, resulting in increased cell viability with decreased apoptosis. THB also reduced the expression of UVB-induced phosphorylated AMP-activated protein kinase (AMPK) and phosphorylated c-Jun N-terminal kinase (JNK). THB suppressed UVB-induced NOX4 expression and ROS generation by inhibiting AMPK and JNK signaling pathways, thereby inhibiting cellular damage. These results showed that THB could be developed as a UV protectant.

• The Korean Journal of Food And Nutrition
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• v.25 no.3
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• pp.564-569
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• 2012

Selenium was initially considered toxic to humans, but it was then discovered that selenium is essential for normal life processes. Selenium plays important roles in antioxidants. It is expected that chitosan microcapsules containing nano-selenium will be able to be used as a key material in bio-medical and cosmetic applications. The high concentration of chitosan derivatives guarantees increased antioxidative activity. Both inorganic and organic forms of selenium can be nutritional sources. The antioxidant properties of selenoproteins help prevent cellular damage from free radicals. The objective of this experiment was to study the antioxidative activity of chitosan nano-selenium. Our experiments were divided into five groups, in the presence of various concentrations(0.1%, 0.3%, 0.5%, 0.7%, and 0.9%) of chitosan. We performed an assessment of the antioxidant properties and cytotoxicity of respective concentrations of chitosan nano-selenium. The antioxidant activity was examined by the free radical scavenging activity on 1,1-diphenyl-2-picrylhydrazyl(DPPH) assay. The cytotoxicity effect was measured by means of 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay. As a result, the electron donating abilities of 0.1%, 0.3%, 0.5%, 0.7%, and 0.9% of chitosan nano-selenium exhibited effective andioxidant scavenging activity at 12.5 ${\mu}g/m{\ell}$ against DPPH radicals. 0.3% chitosan nano-selenium did not show cytotoxicity on human keratinocytes. In general, the cytotoxicity of 0.1% and 0.9% chitosan nano-selenium showed the lowest effects. Though low cytotoxicity of 0.5% and 0.7% chitosan nano-selenium exhibited 29.67% and 38.4% against human keratinocytes on adding 100 ${\mu}g/m{\ell}$ and 50 ${\mu}g/m{\ell}$, respectively, cell vitality was recovered with 200 ${\mu}g/m{\ell}$. These findings support the notion that chitosan nano-selenium may be useful as a new active ingredient source for bioactive compounds.

• Journal of Life Science
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• v.31 no.4
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• pp.418-424
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• 2021

Skin inflammation (dermatitis) is caused by varying skin damage due to ultraviolet radiation and microbial infection. Currently prescribed drugs for dermatitis include anti-histamine and steroid drug classes that soothe inflammation. However, incorrect or prolonged use of steroids can cause weakening of skin barriers as well as osteoporosis. Therefore, treating dermatitis with a drug that has minimal side effects is important. Statins, also known as 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors, are cholesterol-lowering drugs that have been widely treated for hyperlipidemia and cardiovascular diseases. Interestingly, recent studies have shown the anti-inflammatory effects of statins in both experimental and clinical models for of osteoarthritis. This study investigated the possible anti-inflammatory effects of atorvastatin and fluvastatin in human keratinocytes (HaCaT cells), which are crucial components of skin barriers. Stimulation of HaCaT cells with IL-1β increased the expression of the COX2 protein, a major player of inflammatory responses. However, this induction of the COX2 protein was downregulated by pretreatments with atorvastatin and fluvastatin. Treatment with IL-1ß-induced the upregulation of other inflammatory genes (such as iNOS and MMP-1) and these expressions were similarly lowered by these two statin drug treatments. Taken together, these results indicated that atorvastatin and fluvastatin can reduce IL-1β-induced inflammatory responses in HaCaT cells. In conclusion, the findings suggest that atorvastatin and fluvastatin can be potential modulators for ameliorating skin inflammation.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.49 no.1
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• pp.87-96
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• 2023

This study was conducted to discover substances that regulate skin surface acidification using human epidermal keratinocyte cell lines, and to investigate their effects on the moisturizing ability and skin barrier function of the stratum corneum. Prunella vulgaris (P. vulgaris) is an herb widely distributed in Northwest Africa and North America that has been studied for its anti-apoptotic, antioxidant, and anti-inflammatory effects. However, research on the regulation of NHE1 expression and the restoration of skin barrier function has not been conducted. Analysis of P. vulgaris revealed the presence of rosmarinic acid and caffeic acid as active ingredients, which were tested for toxicity in human epidermal keratinocyte cell lines (HaCaT), and showed no toxic effects were observed at high concentarion (100 ㎍/mL or 100 µM). It is known that sodium-hydrogen ion exchange pumps (NHE1) decrease in expression in aging skin to maintain the acidic pH of the stratum corneum, and it is hypothesized that this decrease plays an important role in the impaired restoration of skin barrier function in aging skin. P. vulgaris extract and caffeic acid increased the expression of NHE1 in keratinocytes, increased the expression of natural moisturizing factor (NMF) precursor filaggrin and ceramide synthesis enzyme serine palmitoyl transferase (SPT). In addition, P. vulgaris and caffeic acid decreased the extracellular pH of keratinocytes, indicating a direct effect on skin pH regulation. Taken together, these results suggest that P. vulgaris and caffeic acid can regulate skin pH through NHE1 modulation, and may help to restore skin barrier function by increasing NMF and ceramide synthesis. These results show the possibility that honeysuckle and caffeic acid can have a positive effect on skin health, and can be the basis for the development of new skin protection products using them.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.2
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• pp.151-157
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• 2003

Obtaining a sufficient amount of healthy keratinocytes from a small tissue is difficult. However, ORS cells can be a good source of epithelium since they are easily obtainable and patients do not have to suffer from scar formation at donor sites. Accordingly, the current study modified the conventional primary culture technique to overcome the low propagation and easy aging of epithelial cells during culturing. In a conventional primary culture, the average yield of human ORS tells is 2.↑ $\times$ 10$^3$cells/follicle based on direct incubation in a trypsin (0.1%)/EDTA(0.02%) solution for 15 min at 37$^{\circ}C$, however, our modified method was able to obtain about 6.9 $\times$ 10$^3$cel1s/follicle using a two-step enzyme digestion method involving dispase (1.2 U/mL) and a trypsin (0.1%)/EDTA (0.02%) solution. Thus, the yield of primary cultured ORS cells could be increasd three times higher. Furthermore, a total of 2.0 $\times$ 10$^{7}$ cells was obtained in a serum-free medium. while a modified E-medium with mitomycin C-treated feeder tells produced a total of 6.3 $\times$ 10$^{7}$ Cel1s over 17 days When Starting With 7.5 $\times$ 10$^4$cells. Finally, We Confirmed the effectiveness of our ORS tell isolation method by presenting their ability for reconstructing the bioartificial skin epithelium in vitro

• Journal of Life Science
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• v.25 no.8
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• pp.880-888
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• 2015

Foeniculum vulgare (FV) has long been used in traditional medicine for the treatment of inflammatory diseases. In addition, it is usually known as an important medicinal and aromatic plant widely used as a carminative, digestive, lactogogue, and diuretic, and for treating respiratory and gastrointestinal disorders. The skin barrier protects against the invasion of pathogens, fends off chemical and physical assaults, and protects against extensive water loss. In this study, the effects of solvent-fractionated FV fruits on strengthening the skin barrier and maintaining moisture, as well as their antifungal activity, were investigated in human keratinocyte (HaCaT) cells. The expression of involucrin, loricrin, filaggrin, hyaluronic acid synthase, human β defensin, and cathelicidin genes and proteins was measured by reverse transcription polymerase chain reaction (RT-PCR) and western blotting. The production of hyaluronic acid was determined by enzyme-linked immunosorbent assay (ELISA). The butanol fraction increased the expression of involucrin and filaggrin. Both the ethyl acetate and the butanol fractions increased hyaluronic acid production by promoting the expression of hyaluronic acid synthase-1. Although the antimicrobial peptides were increased by FV crude extract and its fractions, the samples did not show a significant effect compared to the normal group. These results suggest that the butanol fraction of FV could be very useful in cosmetics for the treatment of dermatological diseases.