• Title/Summary/Keyword: Skin barrier protein

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Transdermal Permeation Behavior of FITC-BSA using Microneedle (마이크로니들을 이용한 FITC-BSA의 경피투과 거동)

  • Kim, Yun-Tae;Young, Oh-A;Lee, Jun-Hee;Ahn, Sik-Il;Park, Jong-Hak;Lee, Han-Koo;Khang, Gil-Son
    • Journal of Pharmaceutical Investigation
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    • v.38 no.6
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    • pp.357-363
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    • 2008
  • Penetration rate of large molecule through skin is very low due to the barrier effect of stratum corneum. Novel microneedle treatment device with roll was designed for transdermal delivery of large molecular drugs such as vaccine and protein drugs. The permeation rates of FITC labelled bovine serum albumin (FITC-BSA) as a model protein were determined using modified Franz diffusion cell and hairless mouse skin which were treated by hydrogel or solution containing FITC-BSA. Fluorescent spectrophotometer was used to analyze the concentration of FITC-BSA. Microscope using fluorescent filter was used to capture the image and location of FITC-BSA in the skin. We confirmed that permeation rate of BSA was increased with the treatment by microneedle and was increased by the increasing frequency of treatment. Furthermore, the permeation rate observed from hydrogel treated skin was significantly higher than that from solution treated skin.

Noninvasive Detection of Specific Diagnostic Biomarkers for Atopic Dermatitis

  • Chang, Jeong Hyun
    • Biomedical Science Letters
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    • v.25 no.1
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    • pp.15-22
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    • 2019
  • The diagnosis of atopic dermatitis (AD) includes a test that checks allergen-mediated skin reactions and a method of measuring the total IgE and allergen-specific IgE in blood. However, these test methods are performed directly on the patient, which cause some pain or discomfort. In addition, the skin response test or IgE may result in false negative in about 20% of patients. In the present study, to identify specific biomarkers, HaCaT cells were used as a human keratinocyte that make up the skin, were treated IL-4 and IL-13 for 24 hours to induce a situation similar to keratinocytes in AD patients. In the HaCaT cells, pro-inflammatory cytokine such as IL-5, IL-6, and MCP-1 were increased by IL-4 and IL-13 and skin barrier proteins was reduced by IL-4 and L-13. This results showed that a situation similar to the stratum corneum of an actual patient is induced in HaCaT cells. And then the secretions of Kallikrein (KLK) 5 and KLK7 protease were checked by enzyme-linked immunosorbent assay (ELISA). It was specifically increased by IL-4 and IL-13. This showed that AD-related protease can be detected at the protein level using keratinocytes that can be taken in a non-invasive manner and suggested the possibility of applying it to AD diagnosis.

Skin Hydration Effect of Jeju Lava Sea Water (제주용암해수의 피부 보습 효과 연구)

  • Lee, Sung Hoon;Bae, Il-Hong;Min, Dae Jin;Kim, Hyoung-June;Park, Nok Hyun;Choi, Ji Hae;Shin, Jin Seob;Kim, Eun Ju;Lee, Hae Kwang
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.4
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    • pp.343-349
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    • 2016
  • Many minerals and nutrient salts are abundant in Jeju lava sea water. The objective of this study was to evaluate the skin hydration effects of Jeju lava sea water. The skin barrier serves as a protective barrier that prevents the loss of moisture. The water holding capacity and water transport of the epidermis have been proposed to be important determinants of skin hydration. Jeju lava sea water increased the mRNA expression of filaggrin and caspase-14 which is related to natural moisturizing factor (NMF) formation. Aquaporins 3 (AQP3) are proteins that facilitate the transport of water across cell membranes. Jeju lava sea water increased the mRNA expression and protein expression of AQP3. We employed a skin equivalent model to assess the efficacy of Jeju lava sea water. In a skin equivalent model, Jeju lava sea water increased the CD44 (hyaluronic acid receptor) which is related to skin hydration. From these results, we found out Jeju lava sea water maybe help to skin hydration.

Effect of Oil in Water Nanoemulsion Containing a Mixture of Lactic Acid and Gluconolactone for Skin Barrier Improvement (유산 및 글루코노락톤 혼합물을 함유하는 수중유형 나노에멀젼의 피부장벽개선 효과)

  • Ji-Hye Hong;Young Duck Choi;Gye Won Lee;Young Ho Cho
    • Journal of Life Science
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    • v.33 no.11
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    • pp.905-914
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    • 2023
  • To evaluate the effectiveness of the skin barrier improvement of lactic acid (LA) and gluconolactone (GL), the expression of filaggrin, loricrin, hyaluronic acid (HA), hyaluronan syhthase-2 (HAS2), and aquaporine-3 (AQP3) in keratinocytes, and the moisture content and transepidermal water loss (TEWL) by clinical trials were evaluated. The expression levels of filaggrin and locricrin, which are the main factors affecting the proper functioning of skin barrier function, and HA, HAS2, and AQP3, which are skin moisturizing-related proteins measured by quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting. The results showed that the expression levels of the factors that decreased by H2O2 treatment were significantly increased by LA, GL, and a mixture of LA and GL at the mRNA and protein levels (p<0.05). The nanoemulsion containing a mixture of LA and GL was prepared using the emulsion inversion method, and the average particle size was 299.9 ± 0.287 nm. After measuring the TEWL of nanoemulsion using Vapometer, it was found that TEWL significantly decreased by 15.53% and 26.73% after two weeks and four weeks of product use, respectively, compared to TEWL before product use (p<0.001). Similarly, the skin moisture content of the nanoemulsion significantly increased by 15.40% and 26.59% after two weeks and four weeks of product use, respectively, compared to skin moisture content before product use (p<0.001). Therefore, the skin barrier function and moisturizing effect of a mixture of LA and GL are shown by increasing the moisture content and decreasing the TEWL by increasing the expression of filaggrin, loricrin, HA, HAS2, and AQP3. This suggests the possibility for the development of functional cosmetic ingredients in the future.

Effect of Fritillariae Thunbergii Bulbus for Allergic Inflammation on HaCaT and RBL2H3 Cells (HaCaT 세포와 RBL2H3 세포에서 패모 추출물의 알레르기 염증 완화 효과)

  • Kim, Eun-Young;Lee, Bina;Kim, Jae-Hyun;Hong, SooYeon;Kim, Minsun;Park, Jae Ho;Kim, Jwa-Jin;Sohn, Youngjoo;Jung, Hyuk-Sang
    • The Korea Journal of Herbology
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    • v.34 no.1
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    • pp.23-31
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    • 2019
  • Objectives : The aim of this study was to investigate the effect for allergic-inflammation of Fritillariae Thunbergii Bulbus (FTB) on HaCaT cells and RBL2H3 cells. Methods : To investigate the effects of FTB for anti-inflammation in HaCaT cells, the cells were pretreated with FTB for 1h and then stimulated with $TNF-{\alpha}/IFN-{\beta}$ for 24h. Then thymus and activation-regulated chemokine (TARC) and Macrophage-derived chemokine (MDC) levels were analyzed with ELISA kit. Also to investigate the effect of skin barrier protein, the cells were treated with FTB of various concentrations, and then cells were harvested, expressions of skin barrier protein were measured with RT-PCR. To investigate the effects of FTB for anti-allergy in RBL2H3 cells, the cells were pre-treated with FTB for 1h, and then stimulated with A23187 for 30 min. ${\beta}$-hexosaminidase, IL-4 and $TNF-{\alpha}$ were measured using cultured media. The cells were harvested to analyze the mechanism of the effect for FTB via Western blot. Results : FTB did not show cytotoxicity in HaCaT and RBL2H3. In HaCaT cells, FTB significantly suppressed the expression of TARC, MDC at a dose-dependent manner and markedly increased formation of the skin barrier proteins. In RBL2H3 cells, FTB decreased release of the ${\beta}$-hexosaminidase, IL-4 and $TNF-{\alpha}$ in RBL2H3 through inhibition of the phosphorylation of JNK and p38, which are include in the signaling mechanism of MAPK Conclusion : These results indicate that FTB has an anti-inflammatory effect on the allergic response through blocking MAPK pathway. This suggest that FTB could be a therapeutic agent for allergic response.

Potential Role of Dietary Salmon Nasal Cartilage Proteoglycan on UVB-Induced Photoaged Skin

  • Hae Ran Lee;Seong-Min Hong;Kyohee Cho;Seon Hyeok Kim;Eunji Ko;Eunyoo Lee;Hyun Jin Kim;Se Yeong Jeon;Seon Gil Do;Sun Yeou Kim
    • Biomolecules & Therapeutics
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    • v.32 no.2
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    • pp.249-260
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    • 2024
  • New supplements with preventive effects against skin photodamage are receiving increasing attention. This study evaluated the anti-photoaging effects of salmon nasal cartilage proteoglycan (SPG), acting as a functional material for skin health. We administered SPG to in vitro and in vivo models exposed to ultraviolet B (UVB) radiation and assessed its moisturizing and anti-wrinkle effects on dorsal mouse skin and keratinocytes and dermal fibroblasts cell lines. These results showed that SPG restored the levels of filaggrin, involucrin, and AQP3 in the epidermis of UVB-irradiated dorsal skin and keratinocytes, thereby enhancing the keratinization process and water flow. Additionally, SPG treatment increased the levels of hyaluronan and skin ceramide, the major components of intercellular lipids in the epidermis. Furthermore, SPG treatment significantly increased the levels of collagen and procollagen type 1 by down-regulating matrix metalloproteinase 1, which play a crucial role in skin fibroblasts, in both in vitro and in vivo models. In addition, SPG strongly inhibited mitogen-activated protein kinase (MAPKs) signaling, the including extracellular signal-regulated kinase, c-Jun N-terminal kinase (JNK), and p38. These findings suggest that dietary SPG may be an attractive functional food for preventing UVB-induced photoaging. And this SPG product may provide its best benefit when treating several signs of skin photoaging.

Effect of Mori Follium Extract on the Melanogenesis and Skin Fibril Matrix (상엽(桑葉) 추출물의 미백활성 및 피부섬유구조 형성에 미치는 효과)

  • Kwon, O Jun
    • The Korea Journal of Herbology
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    • v.31 no.5
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    • pp.41-46
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    • 2016
  • Objectives : The Skin is composed of multiple layers, including the epidermis, dermis, and hypodermis. It provides a vital barrier structure that protects vertebrates from external environmental antigens, solvents, ultraviolet light, microorganisms, toxins, and weather conditions. Although several biological effects of Mori Follium have been reported, beneficial effects of Mori Follium in skin health remain unclear. In this study, we prepared water extract of Mori Follium (MLE) and evaluated the effects on melanin accumulation and expression levels of skin fibril-related proteins.Methods : The cytotoxicities of MLE in B16F10 melanoma and human skin fibroblasts (HSF) were examined by MTT assay. Inhibitory effect of MLE on the α-MSH- and IBMX-induced melanosis in B16F10 melanoma was examined. The expression levels of fibronectin, collagen 1α2, and CCN2 in MLE-treated HSF were analyzed by reverse transcription-polymer chain reaction (RT-PCR) and western blotting.Results : The MLE treatment for 24 h did not affect to the B16F10 and HSF at concentrations of 1, 10, 50, 100, 200, 400 and 800 ㎍/ml. The MLE treatment for 72 h significantly and dose dependently suppressed melanin accumulation in B16F10 melanoma. In addition, the MLE treatment up-regulated expression levels of skin fibril-related genes such as fibronectin, collagen 1α2, and CCN2 in HSF. Our western blot analysis revealed MLE-induced up-regulation of skin fibril-related genes required the activation of CCN2 protein.Conclusions : In conclusion, these findings suggest that the MLE could be used in development of cosmetic natural material of maintaining healthy skin.

Skin Moisturizing Properties and Anti-Inflammatory effects of extracts from Coptis chinensis in HaCaT cells (HaCaT cell에서 황련 추출물(Coptis chinensis)의 피부보습과 항염증 효과)

  • Kim, Eun-Hee;Moon, Young-Lan;Jang, Young-Ah
    • Journal of the Korean Applied Science and Technology
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    • v.38 no.3
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    • pp.870-882
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    • 2021
  • Coptis chinensis has been used in the treatment of various diseases such as soothing, anti-inflammation, antimicrobial and antipyretic in oriental traditional medicine. In this study, we investigated the effect of hot water extract of Coptis chinensis(CCW) on skin barrier and inflammation-related factors in UVB and TNF-α/IFN-γ-induced HaCaT cells and evaluated its potential as a moisturizing and anti-inflammatory material. Based on result, the amount of HA (Hyaluronic acid) production and protein and mRNA expression of filaggrin were measured. In TNF-α/IFN-γ-induced HaCaT cells, CCW increased the amount of HA production in a concentration-dependent manner. In the measurement of protein and mRNA expression of filaggrin, the expression rate increased as the concentration of CCW increased. In UVB-induced HaCaT cells, CCW decreased the production of ROS and showed significant results with EGCG ((-)-epigallocatechin-3-gallate), a positive control. In addition, CCW inhibited the expression of inflammatory cytokines TNF-α, IL-1β, IL-6, and IL-8 in TNF-α/IFN-γ-induced HaCaT cells. It was confirmed that the protein and mRNA expression of COX-2, a major factor in skin inflammation, was decreased in a concentration-dependent manner. These results suggest that hot water extract from Coptis chinensis can be used as a cosmetic material having a moisturizing and anti-inflammatory effect.

The Effect of Adiponectin on the Regulation of Filaggrin Expression in Normal Human Epidermal Keratinocytes

  • Choi, Sun Young;Kim, Min Jeong;Ahn, Ga Ram;Park, Kui Young;Lee, Mi-Kyung;Seo, Seong Jun
    • Annals of dermatology
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    • v.30 no.6
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    • pp.645-652
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    • 2018
  • Background: Adiponectin, an adipokine secreted from adipocytes, affects energy metabolism and also shows anti-diabetic and anti-inflammatory properties. Recent studies have reported that adiponectin plays a role in regulating skin inflammation. Objective: This study aimed to investigate the effect of adiponectin on the expression of filaggrin (FLG) in normal human epidermal keratinocytes (NHEKs). Methods: NHEKs were serum-starved for 6h before being treated with adiponectin. Afterward, cell viability was assessed by MTT assay. We also treated with calcium, interleukin (IL)-4, and IL-13 to provide positive and negative comparative controls, respectively. Gene mRNA expression was quantified using real time reverse transcription polymerase chain reaction, and protein expression was evaluated using Western blot. To evaluate the relationship among mitogen-activated protein kinases (MAPKs), activator protein 1 (AP-1), and FLG, we also treated cells with inhibitors for MAPKs JNK, p38, and ERK1/2. Results: FLG and FLG-2 mRNA expression in NHEKs significantly increased after treatment with $10{\mu}g/ml$ adiponectin. Adiponectin also restored FLG and FLG-2 mRNA expression that was otherwise inhibited by treatment with IL-4 and IL-13. Adiponectin induced FLG expression via AP-1 and MAPK signaling. Conclusion: Adiponectin positively regulated the expression of FLG and could be useful as a therapeutic agent to control diseases related to disrupted skin barrier function.

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.