• Title/Summary/Keyword: skin barrier function

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The Expression Pattern of the Tight Junction Protein Occludin in the Epidermal Context When Comparing Various Physical Samples (신체 부위별 표피에서 밀착연접 단백질 중 오클루딘의 발현도 연구)

  • Kim, Ji Sook;Jang, Hyung Seok
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.4
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    • pp.267-272
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    • 2015
  • 'Tight junctions (TJ)' have recently been identified in the granular cell layer of the human epidermis, where they contribute to the normal adhesion between keratinocytes and to the physiologic barrier function of the epidermis. Among the TJ proteins in the epidermis, occludin is an important transmembrane protein, which is considered as a major component. The purpose of this study is to investigate whether regional variation exists in the expression of the tight junction protein occludin in normal human epidermis. Indirect immunofluorescence staining for occludin was performed with specimens taken from different areas of normal skin (4 from each of 7 different anatomical sites, including the scalp, face, posterior neck, upper arm, abdomen, lower back, and inner thigh). The degrees of the expression-intensity in each specimen were estimated with the reciprocals of positive end-point titer of occludin in an indirect immunofluorescence study. The highest degree expression-intensity of the TJ protein occludin among the different areas of normal epidermis was observed on the face and abdomen with a titer of 600 (p=0.001). The lowest intensity of expression of occludin was seen in the epidermis from the upper arm. Skin specimens from the scalp, neck, back, and leg demonstrated intermediate degrees of the expression in intensity. The expression of occludin in the skin samples obtained from different locations of the body showed a statistically significant variation. This suggests that there is a certain degree of regional variation in the expression-intensity of TJ protein 'occludin' in the human epidermis.

Biosynthesis of ceramide by deletion mutant of Saccharomyces cerevisiae (Saccharomyces cerevisiae deletion mutant의 세라마이드 생합성)

  • Kim, Se-Kyung;Noh, Yong-Ho;Yun, Hyun-Shik
    • KSBB Journal
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    • v.24 no.1
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    • pp.25-29
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    • 2009
  • Ceramide is important not only for the maintenance of the barrier function of the skin but also for the water-binding capacity of the stratum corneum. Though the effectiveness of ceramide is not understood fully, ceramide has become a widely used ingredient in cosmetic and pharmaceutical industries. However, ceramide production from Saccharomyces cerevisiae has not been widely studied and the quantity are very low. Gene deletion in the cell is used frequently to investigate the function of gene and verification research of drug target. Specially, deletion mutant library is useful for a large amount functional analysis of gene. In this study, deletion mutants of genes on the metabolic pathway of ceramide synthesis in S. cerevisiae were grown in a batch culture and the cellular content of ceramide was measured. The ceramide content was highest in ${\triangle}$ydc1 mutant and 6 mg ceramide/g cell was obtained.

Phosphatidylserine Enhances Skin Barrier Function Through Keratinocyte Differentiation (포스파티딜세린의 각질세포 분화 유도를 통한 피부장벽 기능 강화)

  • Chung, So-Young;Nam, Sang-June;Choi, Wang-Keun;Seo, Mi-Young;Kim, Jin-Wook;Lee, Seung-Hun;Park, Chang-Seo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.32 no.1 s.55
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    • pp.17-22
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    • 2006
  • Phosphatidyiserine (PS) is a phospholipid which plays the structural role in membranes and serves as a cofactor of signaling enzymes for diverse cellular functions. In this study, we observed that topical treatment with PS significantly decreased trans-epidermal water loss (TEWL) induced by tape-stripping in hairless mice. Also, ceramides in epidermis were increased in PS-treated group compared to vehicle-treated one in vivo. the amounts of non-hydroxyl ceramide (NHCER) (1.4 fold) and glucosylceramide (glucosylCER) (1.6 fold), in the skin of hairless mice, were increased by topical treament with PS. Also, we demonstrated that PS stimulated keratinocyte differentiation. We observed that PS treatment morphologically altered normal human keratinocyte (NHK) from the proliferating phase to the differentiating one, suggesting that PS stimulated epidermal differentiation in NHK. We also showed that the expressions of the specific markers for epidermal differentiation, involucrin (INV) (3.5 fold up) and transglutaminase 1 (TG'ase 1) (3 fold up), were significantly increased by PS treatment, compared to untreated control in vitro. In addition, topical treatment with PS resulted in a progressive increase in INV and loricrin protein levels in vivo. In conclusion, we provide the first evidence for the physiological activities of PS in skin, and we suggest that PS strengthen the epidermal permeability harrier by stimulation of keratinocyte differentiation.

Studies on Antioxidant, Anti-inflammatory and Whitening Effects of Oriental Herbal Extracts (Mix) including Eucommiae cortex (두충을 포함하는 한방추출물(Mix)의 항노화, 항염, 미백 효능 활성에 관한 연구)

  • Choi, Da Hee;Kim, Mi Ran;Kim, Min Young;Kim, Ho Hyun;Park, Sun-Young;Hwang, Hyung Seo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.45 no.1
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    • pp.37-47
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    • 2019
  • Recently, due to the increase in skin diseases caused by particulate matter, endocrine disruptor and environmental changes, the trend of development of cosmetic materials has been shifting to the more safe and effective ingredients based on natural materials rather than existing synthetic compounds like steroids and antihistamines. This study aimed to develop a new natural cosmetic materials using oriental herbs such as Eucommiae cortex, Alpinia oxyphylla Miquel and Bombyx batryticatus. First, DPPH assay was performed to examine the antioxidative activity of the herbal extract (Mix) and 98.8% DPPH radical scavenging activity was confirmed at $400{\mu}g/mL$ concentration of it. In order to confirm the whitening efficacy of oriental herbal extracts(mix), the amount of melanin synthesized after stimulation of ${\alpha}-MSH$ with B16F10 cells was measured. Results showed that it was decreased to 27.1% comparing with the only ${\alpha}-MSH$ treated group, which confirmed the whitening efficacy. Also, both nitric oxide(NO) production and iNOS and COX-2 expression were significantly reduced in RAW264.7 macrophages activated by LPS in the presence of the extracts(Mix). The mRNA expression of the inflammatory cytokines such as $IL-1{\alpha}$, $IL-1{\beta}$, IL-6, and $TNF-{\alpha}$ was also analyzed to confirm the inhibition effect of the extracts on inflammation. Finally, to confirm the enhancement of skin barrier function, the expression of claudin 1 gene, a tight junction protein, was observed using human keratinocyte HaCaT cells and increased as concentration dependent manner. From these results, it is concluded that the oriental herbal extracts(Mix) containing Eucommiae cortex, Alpinia oxyphylla Miquel and Bombyx batryticatus is effective for antioxidant, anti-inflammation, skin whitening, and skin barrier and thus could be applied as a new natural cosmetic material.

Development of Dermal Transduction Epidermal Growth Factor (EGF) Using A Skin Penetrating Functional Peptide (피부투과 기능성 펩타이드를 이용한 경피투과성 상피세포성장인자의 개발)

  • Kang, Jin Sun;La, Ha Na;Bak, Sun Uk;Eom, Hyo Jung;Lee, Byung Kyu;Shin, Hee Je
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.45 no.2
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    • pp.175-184
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    • 2019
  • The epidermal growth factor (EGF) has a intrinsic function of inducing growth and proliferation of cells through interacting with cell membrane receptors in human epidermis and dermis layer. These functions of EGF are used as a main ingredient for wound healing medicines and anti-aging cosmetics. As a cosmetic ingredient, the EGF has a problem in exhibiting its natural efficacy due to the lack of the ability to penetrate through the stratum corneum, which is known as the skin barrier. In this study, a recombinant human epidermal growth factor ($MTD_{151}-EGF$) fused with the macromolecule transduction domain $(MTD)_{151}$ with the skin penetration ability was developed to improve the skin penetration efficiency of the EGF. Expression of $MTD_{151}-EGF$ was performed in E. coli transformed with a vector encoding the $MTD_{151}-EGF$ gene and then purified. The purified $MTD_{151}-EGF$ was evaluated using cell proliferation assay, cytotoxicity test and skin penetration test by franz diffusion cell assay and artificial skin. Cell proliferation activity of $MTD_{151}-EGF$ purified to high purity of 99% or above was equivalent to the EGF or better, and cytotoxicity was not observed. In addition, the $MTD_{151}-EGF$ showed an excellent penetration efficiency compared to the EGF in the skin penetration test with EGF and $MTD_{151}-EGF$ labeled by FITC in an artificial skin penetration model. Based on the quantitative analysis of the penetrating substance using franz diffusion cell assay, the amount of penetration was about 16 times more than that of EGF. These results can be regarded as an effective alternative to improve the existing physical transdermal penetration method related to the use of various active ingredients for cosmetics.

A Literature Study about Comparison of Eastern-Western Medicine on the Acne (여드름의 동(東)·서의학적(西醫學的) 문헌(文獻) 고찰(考察))

  • Joo, Hyun-A;Bae, Hyeon-Jin;Hwang, Chung-Yeon
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.25 no.2
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    • pp.1-19
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    • 2012
  • Objective : The purpose of this study is to investigate about comparison of Eastern-Western medicine on the acne. Methods : We searched Eastern and Western medicine books for acne. We analyzed these books and examined category, definition, etiology, classification, internal and external methods of treatment of acne. Results : The results were as follows. 1. In Eastern medicine, Acne belongs to the category of the Bunja(粉刺), Jwachang(痤瘡), Pyepungbunja(肺風粉刺). In Western medicine, the other name of Acne is acne vulgaris. 2. In Eastern medicine, the definition of Acne includes manual extraction of comedones and skin appearance. In Western medicine, Acne is a common skin disease during adolescence and a chronic inflammatory disease of pilosebaceous unit of self localization. It is characterized by noninflammatory, open or closed comedones and by inflammatory papules, pustules, and nodules and it affects the areas of skin with the densest population of sebaceous follicles, these areas include the face, neck, back, and the upper part of the chest. 3. In Eastern medicine, the cause and mechanism of Acne arose from the state of internal dampness-heat and spleen-stomach internal qi deficiency due to dietary irregularities and then invaded external pathogen such as wind-dampness-heat-cold-fire in lung meridian lead to qi and blood heat depression stagnation. So it appears in skin. In Western medicine, the etiology and pathogenesis of Acne is clearly not identified, but there are most significant pathogenic factors of blood heat depression stagnation. So it appears in skin. In Western medicine, the etiology and pathogenesis of Acne is clearly not identified, but there are most significant pathogenic factors of Acne; Androgen-stimulated production of sebum, hyperkeratinization and obstruction of sebaceous follicles, proliferation of Propionibacterium acnes and inflammation, abnormaility of skin barrier function, genetic aspects, environmental factors etc. 4. In Eastern medicine, differentiation of syndromes classifies clinical aspects, and cause and mechanism of disease; the former is papular, pustular, cystic, nodular, atrophic, comprehensive type; the latter is lung blood heat, intestine-stomach dampness-heat, phlegm-stasis depression, thoroughfare-conception disharmony, heat toxin type. In Western medicine, it divides into an etiology and invasion period, and clinical aspects; Acne neonatorum, Acne infantum, Acne in puberty and adulthood, Acne venenata; Acne vulgaris, Acne conglobata, Acne fulminans, Acne keloidalis. 5. In Eastern medicine, Internal methods of treatment of Acne are divided into five treatments; general treatments, the treatments of single-medicine and experiential description, the treatments depending on the cause and mechanism of disease, and clinical differentiation of syndromes, dietary treatments. In Western medicine, it is a basic principles that regulation on production of sebum, correction on hyperkeratinization of sebaceous follicles, decrease of Propionibacterium acnes colony and control of inflammation reaction. Internal methods of treatment of Acne are antibiotics, retinoids, hormone preparations etc. 6. In Eastern medicine, external methods of treatment of Acne are wet compress method, paste preparation method, powder preparation method, pill preparation method, acupuncture and moxibustion therapy, ear acupuncture therapy, prevention and notice, and so on. In Western medicine, external method of treatments of Acne are divided into topical therapy and other surgical therapies. Topical therapy is used such as antibiotics, sebum regulators, topical vitamin A medicines etc and other surgical therapies are used such as surgical treatments, intralesional injection of corticosteroids, skin dermabrasion, phototherapy, photodynamic therapy, and so on. Conclusions : Until now, there is no perfect, effective single treatment. We think that Eastern medicine approach and treatment can be helpful to overcome the limitations of acne cure.

Brazilin downregulates CCL20 expression via regulation of STAT3 phosphorylation in TNF-α/IL-17A/IFN-γ-induced HaCaT cells (TNF-α/IL-17A/IFN-γ 유도된 HaCaT 세포에서 브라질린의 STAT3 인산화 억제를 통한 CCL20 저해 효과)

  • Kim, Mi Ran;Hwang, Hyung Seo
    • Journal of Applied Biological Chemistry
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    • v.64 no.2
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    • pp.185-192
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    • 2021
  • Psoriasis is a chronic intractable skin disease caused by various inflammatory cytokines such as IL-6, CXCL8, TNF-α, and IFN-γ, as well as IL-17A secreted from Th17 cells and is characterized by hyperkeratosis and chronic inflammation of the epidermis. Brazilin, an active ingredient of Caesalpinia sappan L., is known to exert antioxidant and anti-inflammatory activity, and function in skin barrier improvement. In particular, it was shown as a potential material for treating psoriasis in a tumor necrosis factor (TNF)-α-stimulated HaCaT keratinocyte model. However, the direct regulation of the C-C motif chemokine ligand (CCL) 20, a psoriasis-inducing factor, by brazilin has not been reported. Therefore, in this study, we investigated the suppression of CCL20 and the regulatory mechanism by brazilin using a psoriasis-like model. First, brazilin downregulated CCL20 and CXCL8 in IL-17A-stimulated HaCaT cells in a concentration-dependent manner by inhibiting signal transducer and transcription (STAT)3 phosphorylation. In addition, brazilin significantly inhibited the expression of psoriasis-related genes CXCL8, CCL20, IL-1, IL-6, and TNF-α in TNF-α/IL-17A/IFN-γ-stimulated HaCaT cells. Moreover, brazilin also had a positive effect on improving the skin barrier in TNF-α/IL-17A/IFN-γ-stimulated HaCaT cells. The above results indicated that brazilin ultimately downregulated CCL20 expression by inhibiting STAT3 phosphorylation, and also suppressed the expression of psoriasis-induced cytokines. If the efficacy of brazilin in improving psoriasis is verified through animal models and clinical trials in the future, it may represent a potentially therapeutic substance for psoriasis patients.

Characteristics of Percutaneous Absorption of Glycol ethers (Glycol ethers에 대한 피부 투과 특성)

  • Lee, Han-Seob;Choi, Sung-Boo;Kim, Nac-Joo;Keun, Jang-Hyoun;Hwang, Hyun-Suk;Baek, Jung-Hun;Choi, Jin-Ho;Lee, Ho-Joon
    • Journal of the Korean Applied Science and Technology
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    • v.30 no.1
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    • pp.116-126
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    • 2013
  • Glycol ethers are a group of solvents based on alkyl ethers of ethylene glycol commonly used in paints. These solvents typically have a higher boiling point, together with the favorable solvent properties of lower-molecular weight ethers and alcohols. The word "Glycol ethers" was registered as a United States trademark by Union Carbide Corp. Typically, glycol ethers are found in pharmaceuticals, sunscreens, cosmetics, inks, dyes and water based paints. On the other hand, glycol ethers are used in degreasers, cleaners, aerosol paints and adhesives. Most glycol ethers are relatively water soluble, biodegradable and only a few are considered toxic. Therefore, they are unlikely to pose an adverse risk to the environment. Recent study suggests that occupational exposure to glycol ethers is related to low motile sperm count in men, but the finding has been disputed by others. In this study, skin permeation of 3 types glycol ethers were studied in vitro using matrix such as solvent and detergent. The absorption of glycol ethers[methyl glycol ethers(MC), ethyl glycol ethers(EC) and butyl glycol ethers(BC)] has been measured in vitro through rat skin. Epidermal membranes were set up in Franz diffusion cells and their permeability to PBS measured to establish the integrity of the skin before the glycol ethers were applied to the epidermal surface. Absorption rates for each glycol ethers were determined and permeability assessment made to quantify any irreversible alterations in barrier function due to contact with the esters. Types of glycol ethers in vitro experimental results on MC> EC> BC quickly appeared in the following order: skin permeation was beneficial to the skin permeation small molecular weight, the difference in chemical structure, such as hydrophilic, because with the partition coefficient and solubility mechanisms and passive diffusion to increase the speed at which transmission is considered.

Inhibitory Effect of Steviol and Its Derivatives on Cell Migration via Regulation of Tight Junction-related Protein Claudin 8 (스테비올 및 그 유도체의 세포연접 관련 클라우딘 8 발현 조절을 통한 세포이동 저해효과)

  • Choi, Sun Kyung;Cho, Nam Joon;Cho, Uk Min;Shim, Joong Hyun;Kim, Kee K.;Hwang, Hyung Seo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.4
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    • pp.403-412
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    • 2016
  • The tight junction, one of Intercellular junctions, performs a variety of biological functions by bonding adjacent cells, including the barrier function to control the movement of the electrolyte and water. Recent studies have revealed that unusual expression of tight junction-related genes have been shown to be related in cancer development and progression. Recently, there are many reports that control of tight junction proteins expression is closely related to the skin moisture. In this study, we are focusing on the regulating mechanism of tight junction-associated genes by the steviol and its derivatives. Steviol, used as a sweetner, is known to chemical compound isolated from stevia plant. The MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt) assay was carried out in HaCaT cells (human keratinocyte cell line) in order to determine the cytotoxicity. As a result, while steviol showing cytotoxicity from $250{\mu}M$, steviol derivatives are not cytotoxic more than $250{\mu}M$ concentration. We have observed a change in the tight junction protein via quantitative real-time PCR. Claudin 8 among tight junction proteins is only significantly reduced up to 30% in the presence of steviol. In addition, cell migration was inhibited by steviol, not by stevioside and rebaudioside. Finally, we could observe that steviol, not stevioside and rebaudioside, is able to increase the skin barrier permeability through the transepithelial electric resistance (TEER) measurements. These results suggest that the steviol and its derivatives are specifically acts on the tight junction related gene expression, but steviol derivatives are more suitable as a cosmetic material.

Anti-inflammatory Effects of Rebaudioside A in LPS-stimulated RAW264.7 Macrophage Cells (LPS에 의해 자극된 RAW264.7 대식세포에서 Rebaudioside A의 항염 효과)

  • Cho, Uk Min;Hwang, Hyung Seo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.43 no.2
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    • pp.157-164
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    • 2017
  • Stevia (Stevia rebaudiana) is a perennial plant of the genus Stevia, originated in South America. It stores many forms of glycosides, mainly stevioside and rebaudioside A, in which steviol is the basic structure. Steviol glycosides, widely used as sweeteners, are superior to sugar in sweetness. Recently, it has been reported that steviol glycosides are involved not only in the skin whitening and anti-inflammatory effect but also in enhancing skin barrier function through tight junction regulation. Thus, we examined anti-inflammatory effect of rebaudioside A and tried to identify its potential for improving atopic dermatitis as cosmetic ingredients. To investigate the anti-inflammatory effect, cell viability and mRNA expression level of inflammation-related cytokines were measured using mouse macrophage RAW264.7 cells. Cell counting kit 8 (CCK-8) assay was carried out to measure cell viability and the maximum concentration without cytotoxicity was set to $250{\mu}M$. A quantitative real-time RT-PCR method was used for the study of the inflammatory suppression of rebaudioside A. Rebaudioside A inhibited expression of inducible nitric oxide synthase (iNOS) up to 47% and COX-2 up to 41% compared to LPS treated condition. NO synthesis was decreased by rebaudioside A. Also, mRNA expression of interleukin (IL)-$1{\alpha}$, IL-$1{\beta}$ and IL-6 in LPS-stimulated RAW264.7 cells was decreased to 40%, 45% and 59%, respectively, as a concentration-dependent manner. In conclusion, rebaudioside A inhibited the inflammatory response by regulation of cytokine gene expression. From these results, we expect that steviol glycoside, such as rebaudioside A, can be used as a material for improving atopic dermatitis in the future.