• Biomolecules & Therapeutics
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• v.25 no.3
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• pp.296-307
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• 2017

In spite of frequent usage of primary human foreskin keratinocytes (HFKs) in the study of skin biology, senescence-induced block-age of in vitro proliferation has been a big hurdle for their effective utilization. In order to overcome this passage limitation, we first isolated ten HFK lines from circumcision patients and successfully immortalized four of them via a retroviral transduction of high-risk human papillomavirus (HPV) E6 and E7 oncogenes. We confirmed expression of a keratinocyte marker protein, keratin 14 and two viral oncoproteins in these immortalized HFKs. We also observed their robust responsiveness to various exogenous stimuli, which was evidenced by increased mRNA expression of epithelial differentiation markers and pro-inflammatory genes in response to three reactive chemicals. In addition, their applicability to cytotoxicity assessment turned out to be comparable to that of HaCaT cells. Finally, we confirmed their differentiation capacity by construction of well-stratified three dimensional skin cultures. These newly established immortalized HFKs will be valuable tools not only for generation of in vitro skin disease models but also for prediction of potential toxicities of various cosmetic chemicals.

• Proceedings of the Korean Environmental Health Society Conference
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• 2005.11a
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• pp.166-168
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• 2005

The induction of heme oxygenase-1(HO-1) by ultraviolet(UV) radiation provides a protective defense against oxidative stress, and has been well demonstrated in UVA-irradiated skin, but not UVB. In this study in mice, we show that the UVB(290-320nm) radiation can be attributed to the induction of cutaneous heme oxygenase-1. The expression of HO-1 mRNA was assessed in vivo by the reverse transcription-polymerase chain reaction (RT-PCR) analysis, and HO-1 enzyme activity was measured in microsomal preparation from irradiated mice. The mRNA level of HO-1 increases in liver and skin from 24h to 72h after UVB($3KJ/m^3$) radiation. The results of gene expression were same pattern of HO enzyme activity in skin, but not in liver. HO-1 mRNA in liver resulted in a progressive increase to 96h after UVB radiation, but HO activity in liver increased to 48h. This finding indicates that UVB radiation is an important inducer of HO-1 and increases in HO activity may protect tissues directly or indirectly from oxidative stress.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.18 no.1
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• pp.135-153
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• 2005

Herbal mixture water extract of (Phellodendron amurense, Scuellaria baiklensis, Spphora flavescens, Lithospermum erythrorhizon, Mellaphis chinesis, Alumite, Zanthoxylum schinifolium, Glycyrrhiza uralensis), which exhibit several beneficial effects including acne and skin diseases, was tested for anti-microbial activity and anti-inflammation effects. The herbal mixture extract showed antimicrobial activity against Stapylococcus epidermis, Propionibacterium acne, and Malassezia furfur. The growth of Stapylococcus epidermis, Propionibacterium acne, and Malassezia furfur was inhibited allergy and LPS induced cyclooxygenase-2(COX-2)gene expression in RAW24.7macrophage. The results indicated th ear swelling and histamine release induced by compound 48/80 were dose-dependently reduced, ranging 11-38% and 11-56%, respectively. Furthermore the extract inhibited the expression of LPS-induced COX-2 proteins and mRNAs without an appreciable cytotoxic effects on RAW264.7 cells. The cytotoxicity of the extract using M7T assay showed the cytotoxicity of 7 and 18% against L929 cell line. Based on these results, it is concluded that the herbal mixture water extract can be applied to the acne and skin diseases therapy.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.18 no.1
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• pp.154-171
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• 2005

Herbal mixture water extract of (Rheum coreanum Scutellaria baikalensis, Phellodendron amurese), which exhibit several beneficial effects including acne and skin diseases, was tested for anti-microbial activity and anti-inflammation effects. The herbal mixture extract showed antimicrobial activity against Stapylococcus epidermis and Propionbacterium acne. The growth of Stapylococcus epidermis and Propionibacterium acne was inhibited completely by addition of 1.0% of the extract. Also in the present study we examined the mixture extract on compound 48/80 induced allergy and LPS induced cyclooxygenase-2(COX-2) gene expression in RAW264.7 macrophage. The results indicated the ear swelling and histamine release induced by compound 48/80 were dose-dependently reduced, ranging 18-36% and 10-61%, respectively. Furthermore the extract inhibited the expression of LPS-induced COX-2 proteins and mRNAs without an appreciable cytotoxic effects on RAW264.1 cells. The cytotoxicity of the extract using MTT assay showed the cytotoxicity of 6% and 13% against L929 cell line. Based on these results, it is concluded that the herbal mixture water extract can be applied to this acne and skin diseases therapy.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.22 no.2
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• pp.104-117
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• 2009

Objective : Melanin is one of the most important facor in skin color. Melanin protects human skin from ultraviolet radiation otherwise it causes melanin pigmentation. So this experiment is carried out for test whether Scutellaria baicalensis Georgi(SBG) inhibits melanin synthesis and tyrosinase activity in mouse B16 melanoma cells. Method : The melanin synthesis inhibition effects of SBG were examined by in vitro melanin production assay. We assessed inhibitory effects of SBG on melanin contents from B16F1 melanoma cell, on tyrosinase activity(cell and cell free system), effect of SBG on the expression tyrosinase, Microphthalmia-associated Transcription Factor(MITF), Extracellular signal-regulated Kinase(ERK). Result : SBG inhibited melanin synthesis induced $\alpha$-MSH($\alpha$-Melanin Stimulating Hormone) in B16F1. SBG inhibited tyrosinase activity and expression. And SBG down-regulates MITF and stimulated ERK activation in B16F1. Conclusion : According to above results, SBG was improved its suppression effect to the inhibition of melanin synthesis, tyrosinase activation, and tyrosinase promotor activation. So SBG is considered to be used for an strong source of skin whitening effect.

• 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]

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.28 no.1
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• pp.68-84
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• 2015

Object : The object of this study was effected of composed of 4 herbal medicines(Rubus coreanus, Rehmanniae Radix, Houttuynia cordata, Betulae cortex) on acute atopic dermatitis mice. Methods : BALB/c mice were divided 4 groups ; normal group, negative control group of acute atopic dermatitis mice induced by DNCB, treated apply to the back skin with the herbal medicine group, and treated orally with herbal medicine group. Treated with herbal medicine and DNCB during 2 weeks. After treated, measured WBCs, RBCs, and platelet in the blood, and analysed mRNA expression of spleen. Result : Composed herbal medicines could reduce WBCs, lymphocytes, monocytes, neutrophils, and eosinophils in the mouse blood. And, could down TNF-${\alpha}$ levels in spleen. In the skin tissue histology results, herbal medicines reduce the cells and T cells. Conclusion : Composed herbal medicines help to another inflammatory disease because they reduce the cells and T cells in the skin tissues. And they have inhibitory effects of TNF-${\alpha}$ mRNA expression, and Th 2 immune responses. Therefore herbal medicines use for an acute atopic dermatitis treatment.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.26 no.1
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• pp.75-81
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• 2013

Objectives: Hyaluronic acid, high molecular glycosaminoglycan, exists in extracellular matrix of tissue, especially, in skin and has been known to be deeply involved in skin hydration. In this study, we investigated the effect of methanol extract of Hwang-gi, Astragalus membranaceus root, on hyaluronic acid production in human keratinocyte HaCaT cells. Methods: We determined hyaluronic acid synthase 2 gene expression and hyaluronic acid production in HaCaT cells by using RT-PCR and ELISA, respectively. Results: Hwang-gi extract didn't show the toxicity to HaCaT cells within the treated concentration and increased the hyaluronic acid synthase 2 gene expression and hyaluronic acid production. Conclusions: Hyaluronic acid production increased by Hwang-gi could be, partially, contribute to the moisturing effect in skin by it.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.31 no.1
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• pp.32-41
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• 2018

Objectives : Hyaluronic acid(HA) is a mucopolysaccharide, occuring naturally in living organisms. It is one of the most hydrophilic molecules, so it has been known as being related to skin hydration and skin aging. The purpose of this study is to examine the effects of Angelica gigas(A. gigas) ethanol extract on hyaluronic acid synthesis. Methods : To determine cytotoxicity and hyaluronic acid synthase 2 gene expression, hyaluronic acid production in HaCaT cells, MTT assay and RT-PCR ELISA was used. Results : There were no cytotoxicity in $50{\mu}g/ml$ concentration A. gigas extract in MTT assay. Hyaluronic acid synthase 2(HAS2) gene expression was increased by all treated concentration A. gigas extract. Hyaluronic acid production was higher than control group in $50{\mu}g/ml$ & $100{\mu}g/ml$ concentration A. gigas extract. Conclusions : Hyaluronic acid production was increased by A. gigas extracts. Therefore, We suggest that A. gigas can make a contribution to the moisturizing effect on human skin.

• Journal of Animal Reproduction and Biotechnology
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• v.36 no.3
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• pp.137-144
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• 2021

In this study, to analyze whether artificial regulation of apoptosis in the development of somatic cells can affect the stable growth and development of cells, 20 alpha-hydroxysteroid dehydrogenase (20α-HSD) and rapamycin were treated to induce apoptosis and autophagy in the both skin and muscle cells. Respectively, and 3-methyladenine was supplemented to inhibit cell death. Our results show that stimulation with rapamycin activated autophagy in both types of cells, but increased apoptosis more than autophagy in the case of skin cells. These results indicate that there was a difference in the expression of survival factors and cell development depending on the type of cell. In particular, in the expression of autophagy-related gene (MAP1LC3A) was higher than that of Casp-3, an apoptosis factor. Furthermore, cell development was the highest in all cell groups cultured by artificially inducing autophagy, however the lowest in the apoptosis-inhibited group. Especially, the noteworthy result of this study was that when apoptosis was induced using 20α-HSD, it was possible to induce apoptosis in both skin and muscle cells. Therefore, the main point of this study is that apoptosis induced during cell culture plays a pivotal role in cell remodeling.