• Title/Summary/Keyword: mRNA 발현

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Oxidative Stress Inhibitory Effects of Low Temperature-Aged Garlic (Allium sativum L.) Extracts through Free Radical Scavenging Activity (저온숙성마늘의 라디칼 소거 활성을 통한 산화스트레스 억제 효과)

  • Hwang, Kyung-A;Kim, Ga Ram;Hwang, Yu-Jin;Hwang, In-Guk;Song, Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.1
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    • pp.27-34
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    • 2016
  • Garlic has drawn attention as a food material for its anti-oxidative and anti-inflammatory properties as well as for prevention and treatment of cancer. In order to increase efficiency, various aging methods for garlic have been attempted. In particular, thermally processed garlic is known to have higher biological activities due to its various chemical changes during heat treatment. Therefore, in this study, we investigated the anti-oxidative effects of garlic extracts aged at low temperature ($60{\sim}70^{\circ}C$). In the results, 2,2-diphenyl-1-picrylhydrazyl and 2,2-azino-bis (3-ethylbenzo-thiazoline-6-sulfonate) radical scavenging activities and ferric reducing ability of low temperature-aged garlic (LTAG) were similar to those of raw garlic. LTAG also showed decreased lipopolysaccharide (LPS)-induced production of reactive oxygen species, although there were not significant differences among samples. In addition, xanthine oxidase activity was inhibited by LTAG; the 15 days and $60^{\circ}C$ extract showed outstanding inhibition compared with the others. To understand the molecular mechanisms behind the anti-oxidative activity of LTAG, we performed quantitative real-time PCR analysis. The 30 days and $70^{\circ}C$ extract upregulated mRNA expression of antioxidant enzymes such as Cu/Zn-superoxide dismutase (SOD), Mn-SOD, glutathione peroxidase, and catalase in LPS-stimulated RAW 264.7 cells. This result indicates that LTAG can be a functional food as a nature antioxidant and antioxidant substance.

$CH_4-H_2-N_2 $ 기체계에서 MW-PACVD를 이용한 결정상의 합성

  • 김도근;백영준;성태연
    • Proceedings of the Korean Vacuum Society Conference
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    • 1999.07a
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    • pp.54-54
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    • 1999
  • 다이아몬드 합성시 질소 첨가는 Cn 화합물의 합성가능성을 비롯하여 다이아몬드의 질소 도핑, 성장 속도 및 결정성 변화 등 다양한 관점에서 중요한 의미를 가지고 있다. 본 연구에서는 다이아몬드의 일반적인 합성조건에서 질소를 첨가하여 합성된 막의 형상 및 상 변화에 대해 고찰하였다. 막은 다이아몬드 전처리시킨 Si 기판위에 microwave plasma CVD 장치를 이용하여 합성하였다. 유입되는 혼합가스(CH4+H2+N2)에서 N2 첨가량을 0-95%까지 변화시켰다. 이때 CH4 농도는 5%로 고정하였고, 합성온도는 90$0^{\circ}C$-115$0^{\circ}C$까지 변화시켰다. 이와 같이 합성된 막의 표면조직 및 성장 두께를 측정하기 위해 주사전자현미경을 이용하였다. 상의 분석은 Raman, XRD 및 TEM 분석을 이용하였으며, 조성분석을 위해 XPS 및 AES를 사용하였다. 질소 첨가량에 따라 합성된 막은 첨가하지 않은 경우에 다이아몬드 결정에서 시작하여 질소첨가에 따라 결정면이 깨지는 것으로 나타났다. 그러나 30%, 45%의 경우는 다시 결정면이 나타났다. 다량의 질소가 첨가되었을 때, 다시 결정면을 보이는 다이아몬드가 합성된 것은 매우 흥미로운 결과이다. 한편 질소와 메탄만의 기체하에서는 다시 결정면이 관찰되지 않았다. 이들 상의 구조는 XRD 및 TED 분석을 통해 모두 다이아몬드로 확인되었다. 기체내의 질소의 첨가에 관계없이 고상내에 질소는 확인되지 않았다. 따라서 이방법에 의한 CN 화합물의 합성은 힘든 것으로 보여진다. 이들 실험 결과를 근거로 온도 및 조성에 따른 기체의 열역학적 계산을 통하여 합성거동과의 연관성을 검토하였다. anode는 매우 높은 충전용량을 갖는데 첫 번째 방전시에 Li2O를 생성하여 비가역적 반응을 나타내고 계속되는 충방전 동안 Li-Sn 합금이 생성되어 2차전지의 가역적 반응을 가능하게 한다. SnO2 는 대기중에서 Li 금속보다 안정하기 때문에 전지의 제작 공정 및 사용 면에서 매우 우수한 물질이지만 아직까지 SnO2 구조적 특성과 전지의 충, 방전 특성에 대한 관계의 규명을 위한 정확한 정설은 제시되고 있지 못하다. 본 연구에서는 TFSB anode 물질로써 SnOx박막을 상온에서 여러 전도성 콜렉터 위에 증착하여 그 충, 방전 특성을 보고하였다. 증착된 SnOx박막의 표면은 SEM, AFM으로 분석하였으며 구조의 분석은 XR와 Auger electron spectroscope로 하였다. 충, 방전 특성을 분석하기 위하여 리늄 foil을 대극과 참조 전극으로 하여 EC:DMC=1:1, 1M LiPF6 액체 전해질을 사용한 Half-Cell를 구성하여 100회 이상의 정전류 충, 방전 시험을 행하였다. Half-Cell test 결과 박막의 구조, 콜렉터의 종류 및 Sn/O비에 따라 서로 다른 충, 방전 거동을 나타내었다.다. 거의 없었다. 5mTorr 일 때가 가장 좋았다.수 있음을 알 수 있었다. 그러므로, RNA바이러스의 하나인 BVDV의 viral replicon을 이용하여 다양한 종류의 포유동물 세포에 유전자 발현벡터로써 사용할 수 있음으로 post-genomics시대에 다양한 종류의 단백질 기능연구에 맡은 도움이 되리라 기대한다.다양한 기능을 가진 신소재 제조에 있다. 또한 경제적인 측면에서도 고부가 가치의 제품 개발에 따른 새로운 수요 창출과 수익률 향상, 기존의 기능성 안료를 나노(na

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CONSTRUCTION OF HNGF-$\beta$ RECOMBINANT ADENOVIRUS & SCREENING OF ITS EXPRESSION AFTER TRANSFECTION INTO VARIOUS CELL LINES (말초신경재생을 위한 hNGF-$\beta$ recombinant Adenovirus의 제작 및 수종세포주에서 신경성장인자의 발현)

  • Gao, En-Feng;Chung, Hun-Jong;Ahn, Kang-Min;Kim, Yoon-Tae;Park, Hee-Jung;Sung, Mi-Ae;Kim, Nam-Yeol;Yoo, Sang-Bae;Myoung, Hoon;Hwang, Soon-Jung;Kim, Myung-Jin;Kim, Sung-Min;Jang, Jeong-Won;Lee, Jong-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.27 no.5
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    • pp.446-456
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    • 2005
  • Nerve growth factor(NGF) has a critical role in peripheral nerve regeneration. The aim of this study is to construct a well-functioning hNGF-$\beta$ recombinat adenovirus for the ultimate development of improved method to promote peripheral nerve regeneration with adenovirus mediated hNGF-$\beta$ gene transfection into Schwann cells. First PCR associated cloning of GFP-tagged hNGF-$\beta$ which was ligated into E1/E3 deleted adenoviral vector was performed and tranfected into E. coli to construct hNGF-$\beta$ recombinant adenovirus. After production of recombinat adenovirus in a large scale, its transfection efficiency, expression, and function were evaluated using cell lines or primarily cultured cells of HEK293 cells, Schwann cells, fibroblast(NIH3T3) and myocyte(CRH cells). GFP expression was observed in 90% of infected cells compared to uninfected cells. Total mRNA isolated from hNGF-$\beta$ recombinat adenoviru infected cells showed strong RT-PCR band, however, LacZ recombinant adenovirus infected or uninfected cells did not. NGF quantification by ELISA showed a maximal release of 18.865 +/- 0.31ng/mL at 4th day. PC-12 cells exposed to media with hNGF-$\beta$ recombinant adenovirus infected Schwann cell demonstrated higher levels of differentiation compared with controls. We generated hNGF-$\beta$ recombinant adenovirus and induced over expression of NGF successfully in nonneuronal and neuronal cells. Following these result, it is expected to develop an improved treatment strategy peripheral nerve regeneration using the hNGF-$\beta$ gene transfected cells.

Increased osteopontin expression in activated glial cells in experimental autoimmune encephalomyelitis (자가면역성 뇌척수염 흰쥐의 활성화된 신경아교세포에서 증가된 osteopontin의 발현)

  • Park, Suk-jae;Hwang, In-sun;Kim, Gyu-beom;Shin, Tae-kyun;Jee, Young-heun
    • Korean Journal of Veterinary Research
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    • v.46 no.3
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    • pp.177-184
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    • 2006
  • Experimental autoimmune encephalomyelitis (EAE) is a disease model of multiple sclerosis (MS) that is characterized by remittance and relapse of the disease and autoimmune and demyelinating lesions in the central nervous system (CNS). Autoimmune inflammation is maintained by secretion of a large number of protein. Previous studies have suggested that transcripts encoding osteopontin (OPN) are frequently detected in the mRNA population of MS plaques. To elucidate the functional role of OPN in initiation and development of EAE, we examined the expression and localization of OPN in the spinal cord during acute EAE. We demonstrated that OPN significantly increased at the early stage of EAE and slightly declined thereafter by western blot analysis. An immunohistochemical study revealed that OPN was constitutively expressed in some glial cells (microglia, astrocytes) of white matter and neurons in the CNS of control rats. OPN expression was shown to be increased in the same cells at the early and peak stage of EAE. To identity cells expressing OPN by double-immunofluorescence labeling, we labeled rat spinal cord sections for OPN with a monoclonal OPN antibody and with mAbs for astrocyte (GFAP), microglia/macrophage (OX42)-specific markers. The major cell types of OPN-expressing cells were activated astrocytes and microglia in the adjacent inflammatory lesions. Interestingly, OPN was mainly expressed in the end feet of astrocytes around vascular cell adhesion molecule-1 (VCAM-1) expressing endothelial cells of CNS blood vessel. These findings suggest that increased levels of OPN in activated glial cell may play an important role in the recruitment of inflammatory cells into the CNS parenchyma during EAE.

SREBP as a Global Regulator for Lipid Metabolism (지질대사 조절에서 SREBP의 역할)

  • Lee, Wonhwa;Seo, Young-kyo
    • Journal of Life Science
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    • v.28 no.10
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    • pp.1233-1243
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    • 2018
  • Sterol regulatory-element binding proteins (SREBPs) are a family of transcription factors that regulate lipid homeostasis and metabolism by controlling the expression of enzymes required for endogenous cholesterol, fatty acid (FA), triacylglycerol, and phospholipid synthesis. The three SREBPs are encoded by two different genes. The SREBP1 gene gives rise to SREBP-1a and SREBP-1c, which are derived from utilization of alternate promoters that yield transcripts in which distinct first exons are spliced to a common second exon. SREBP-2 is derived from a separate gene. Additionally, SREBPs are implicated in numerous pathogenic processes, such as endoplasmic reticulum stress, inflammation, autophagy, and apoptosis. They also contribute to obesity, dyslipidemia, diabetes mellitus, and nonalcoholic fatty liver diseases. Genome-wide analyses have revealed that these versatile transcription factors act as important nodes of biological signaling networks. Changes in cell metabolism and growth are reciprocally linked through SREBPs. Anabolic and growth signaling pathways branch off and connect to multiple steps of SREBP activation and form complex regulatory networks. SREBPs are activated through the PI3K-Akt-mTOR pathway in these processes, but the molecular mechanism remains to be understood. This review aims to provide a comprehensive understanding of the role of SREBPs in physiology and pathophysiology at the cell, organ, and organism levels.

Effect of Piryongbanggamgil-tang on Airway Mucin Secretion, Production, Gene Expression and Hypersecretion of Mucus (필용방감길탕이 기도 뮤신의 분비, 생성, 유전자 발현 및 점액 과다 분비에 미치는 영향)

  • Kim, Yoon Young;Min, Sang Yeon;Kim, Jang Hyun
    • The Journal of Pediatrics of Korean Medicine
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    • v.28 no.2
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    • pp.56-71
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    • 2014
  • Objectives In this study, the author tried to investigate whether piryongbang-gamgil-tang (PGGT) significantly affect in vitro airway mucin secretion, PMA- or EGF- or TNF-${\alpha}$-induced MUC5AC mucin production / gene expression from human airway epithelial cells and increase in airway epithelial mucosubstances and hyperplasia of tracheal goblet cells of rats. Materials and Methods For in vitro experiment, confluent RTSE cells were chased for 30 min in the presence of PGGT to assess the effect of PGGT on mucin secretion by enzyme-linked immunosorbent assay (ELISA). Also, effect of PGGT on PMA- or EGFor TNF-${\alpha}$-induced MUC5AC mucin production and gene expression from human airway epithelial cells (NCI-H292) were investigated. Confluent NCI-H292 cells were pretreated for 30 min in the presence of PGGT and treated with PMA (10 ng/ml) or EGF (25 ng/ml) or TNF-${\alpha}$ (0.2 nM) for 24 hrs, to assess both effect of PGGT on PMA- or EGF- or TNF-${\alpha}$-induced MUC5AC mucin production by ELISA and gene expression by reverse transcription-polymerase chain reaction (RT-PCR). For in vivo experiment, the author induced hypersecretion of airway mucus and goblet cell hyperplasia by exposure of rats to $SO_2$ during 3 weeks. Effect of orally-administered PGGT during 2 weeks on increase in airway epithelial mucosubstances from tracheal goblet cells of rats and hyperplasia of goblet cells were assesed by using histopathological analysis after staining the epithelial tissue with alcian blue. Possible cytotoxicities of PGGT in vitro were assessed by examining LDH release from RTSE cells and the rate of survival and proliferation of NCI-H292 cells. In vivo liver and kidney toxicities of PGGT were evaluated by measuring serum GOT/GPT activities and serum BUN/creatinine concentrations of rats after administering PGGT orally. Results (1) PGGT did not affect in vitro mucin secretion from cultured RTSE cells. (2) PGGT significantly inhibited PMA-, EGF-, and TNF-${\alpha}$-induced MUC5AC mucin productions and the expression levels of MUC5AC mRNA from NCI-H292 cells. (3) PGGT decreased the amount of intraepithelial mucosubstances and showed the tendency of expectorating airway mucus already produced. (4) PGGT increased LDH release from RTSE cells. However, PGGT did not show in vivo liver and kidney toxicities and cytotoxicity to NCI-H292 cells. Conclusion The result from this study suggests that PGGT can regulate the production and gene expression of airway mucin observed in diverse respiratory diseases accompanied by mucus hypersecretion and do not show in vivo toxicity to liver and kidney functions after oral administration. Effect of PGGT with their components should be further studied using animal experimental models that reflect the diverse pathophysiology of respiratory diseases through future investigations.

The Scaffolding Protein WAVE1 Associates with Kinesin 1 through the Tetratricopeptide Repeat (TPR) Domain of the Kinesin Light Chain (KLC) (Kinesin Light Chain (KLC)의 Tetratricopeptide Repeat (TPR) 도메인을 통한 Scaffold 단백질 WAVE1과 Kinesin 1의 결합)

  • Jang, Won Hee;Jeong, Young Joo;Urm, Sang-Hwa;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.26 no.8
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    • pp.963-969
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    • 2016
  • Kinesin superfamily proteins (KIFs) are microtubule-dependent molecular motor proteins essential for the intracellular transport of organelles and protein complexes in cells. Kinesin 1 is a member of those KIFs that transport various cargoes, including organelles, synaptic vesicles, neurotransmitter receptors, cell signaling molecules, and mRNAs through interaction between its light chain subunit and the cargoes. Kinesin light chains (KLCs) are non-motor subunits that associate with the kinesin heavy chain (KHC) dimer. KLCs interact with many different binding proteins, but their particular binding proteins have not yet been fully identified. We used the yeast two-hybrid assay to identify proteins that interact with the tetratricopeptide repeat (TPR) domain of KLC1. We found an interaction between the TPR domain of KLC1 and Wiskott-Aldrich syndrome protein family member 1 (WAVE1), a member of the WASP/WAVE family involved in regulation of actin cytoskeleton. WAVE1 bound to the six TPR domain-containing regions of KLC1 and did not interact with KHCs (KIF5A, KIF5B, and KIF5C) in the yeast two-hybrid assay. The carboxyl (C)-terminal verprolin-cofilin-acidic (VCA) domain of WAVE1 is essential for interaction with KLC1. Also, other WAVE isoforms (WAVE2 and WAVE3) interacted with KLC1 in the yeast two-hybrid assay. When co-expressed in HEK-293T cells, WAVE1 co-localized with KLC1 and co-immunoprecipitated with KLC1 and KIF5B. These results suggest that kinesin 1 motor protein may transport WAVE complexes or WAVE-coated cargoes in cells.

Effects of anti-inflammatory on Perilla frutescens var. crispa Induced by mutants with γ-Ray (감마선을 이용한 육종 차조기의 항염증 효과)

  • Sim, Boo-Yong;Park, Jung-Hyun;Kim, Sung-Kyu;Ji, Joong-Gu
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.2
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    • pp.488-497
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    • 2019
  • The purpose of this study was to confirmed anti-inflammatory effect the apple Induced by mutants with ${\gamma}-Ray$ extract. Cell viability was assessed by MTT assay using RAW 264.7 cells. The extracts measured through changes in the levels of reactive oxygen species (ROS), nitric oxide (NO), inflammatory cytokines, NF-kB, and COX-2 on LPS-induced RAW 264.7 cells. All test results were analyzed by ELISA reader, Luminex and RT-PCR. In result, the extracts was not toxic below in 25 ug/ml, and extracts was inhibited the productions nitric oxide, ROS, cytokines (IL-1b, IL-6, TNF-a), NF-kB and COX-2 in LPS-induced RAW 264.7 cells. Also, the expression levels were decreased on mRNA of $NF-{\kappa}B$ and COX-2. In other words, Perilla frutescens var. crispa Induced by mutants with ${\gamma}-Ray$ extracts showed significant anti-inflammatory effect. These results may be developed as a raw material for new health food and therapeutics to ease the related to the above mediators.

Inhibitory Effect of Purified Bee Venom(Apis mellifera L.) on Adipogenesis in Korea (국내 양봉농가에서 채취한 정제봉독(Apis mellifera L.)의 지방세포 분화 억제 효과)

  • Han, Sang Mi;Kim, Hyo Young;Woo, Soon Ok;Kim, Se Gun;Choi, Hong Min;Moon, Hyo Jung
    • Journal of Apiculture
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    • v.35 no.1
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    • pp.49-54
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    • 2020
  • Bee (Apis mellifera L.) venom is used for the treatment of various human diseases due to its known anti-inflammatory and antibacterial properties. This study investigated the effect of purified bee venom (PBV) on adipogenesis in 3T3-L1 preadipocytes. There was no cytotoxicity while cells were treated with PBV by various concentrations. In the PBV treated cells increases in fat storage were inhibited and also confirmed by oil red o staining. To understand the underlying mechanism at the molecular level were examined on the expression of the genes involved in adipogenesis by using real-time PCR. In this cell model, the mRNA level of adipogenic genes such as peroxisome-proliferator-activated receptors gamma (PPARγ) and CAAAT/enhancer binding protein alpha(C/EBPα) were decreased by PAE treatment, comparing with those of control group. Theses results suggest that PBV inhibits adipocyte differentiation in 3T3-L1 cells and can be used as an efficient natural substance to manage anti-obesity.

Study on the Whitening Efficacy and Skin Barrier by Lysosome-related Organelle Extract (LOE) from Egg White (난백(Egg White)에서 추출한 리소좀 추출물(LOE)의 미백 효능 및 피부장벽에 관한 연구)

  • Choi, Da Hee;Jeon, Gyeongchan;Yoon, Jihee;Min, Jiho;Park, Si Jun;Kim, Jung Su;Hwang, Ee Taek;Hwang, Hyung Seo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.45 no.4
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    • pp.389-397
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    • 2019
  • Lysosomes are cellular organelles involved in energy metabolism and intracellular digestion in eukaryotic cells, including protease, nuclease, glycosidase, lipase, and phosphatase. Our previous studies have confirmed that egg white lysosomes had melanin decolorization and reduction activity. However, there have been few studies on skin barrier and skin regeneration as well as inhibition of melanin production by egg white lysosomes on B16F10 melanocyte cell line. In this study, we attempted to identify the effect of lysosome-related organelle extract (LOE) extracted from egg white on the melanin content change and skin barrier enhancement in cells. First, cytotoxicity evaluation was performed on B16F10 melanocyte cell line to confirm the whitening efficacy of LOE. Cytotoxicity by LOE was not observed at 20 mg/mL concentration, but cytotoxicity was observed at 40 mg/mL, and the maximum concentration value was set to 20 mg/mL in all subsequent experiments. LOE samples of 5, 10, 20 mg/mL inhibited melanin production by 61.5 ± 4.0%, 61.4 ± 7.3%, 58.3 ± 8.3%, respectivly, compared to α-MSH, a negative control in melanin contents assay. MITF mRNA expression was reduced by about 39.7 ± 3.2% compared to the α-MSH treatment group. TEER assay using HaCaT showed that LOE increased TEER resistance in a dose-dependent manner, indicating that LOE is involved in strengthening the skin barrier. LOE also increased the TEER resistance under TNF-α treatment. Skin barrier was normally restored by LOE even under the condition of inflammation. LOE had a positive effect on cell division and cell migration promotion, confirmed by the observing the effect of promoting cell migration by LOE through cell migration assay. Taken together, we expect that LOE can be developed as a cosmetic material to enhance has effects on skin regeneration and skin barrier strengthening as well as whitening function if enzyme stabilization and formulation technology are combined.