• Title/Summary/Keyword: Tight Junction

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Investigation of the effect of Hibiscus sabdariffa L. extracts on tight-junction related genes in human keratinocyte HaCaT cells (히비스커스 추출물이 인간 각질 형성 세포의 밀착 연접 관련 유전자 발현에 미치는 영향 연구)

  • Jung, Haesoo;Kim, Eunmi;Han, Hyosang;Kim, Keekwang
    • The Korea Journal of Herbology
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    • v.36 no.5
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    • pp.59-67
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    • 2021
  • Objectives : Hibiscus (Hibiscus sabdariffa L.) is rich in antioxidants such as flavonoids and anthocyanins and is known to have anti-inflammatory activity and anti-aging function of the skin, but there is no study on its effect on the skin barrier. This study aim to investigate the positive effect on the skin barrier by confirming the effect of water extracts of Hibiscus sabdariffa L. (WEHS) on the tight junction-related gene expression. Methods : The antioxidant efficacy of WEHS was investigated through ABTS and DPPH assays, and 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium was performed to examine the effect on cell viability. quantitative Reverse transcription polymerase chain reaction and immunoblot analysis were performed to confirm the effect of WEHS on the expression of tight junction-related genes, and immunofluorescence microscopy was used to confirm the movement of Claudin 1 protein into tight junctions. Results : WEHS showed strong antioxidant activity and induced an increase in both mRNA and protein expression levels of Claudin 1 among tight junction-related genes. The strong localization of Claudine 1 protein increased by WEHS to the tight junction was confirmed by immunofluorescence microscopy. Conclusions : Hibiscus was confirmed through this study to show antioxidant activity and the function of promoting the expression of the tight junction Claudin 1 gene, suggesting that Hibiscus can be used as a material for the prevention and treatment of skin moisturizing and atopy, which have an important influence on tight junction.

Inhibitory effect of ethanol extract of Codonopsis lanceolata against oxidative stress and disruption of tight cell junction in NCI-H441 cells after exposure to urban particulate matter (더덕 에탄올 추출물의 도시미세먼지 노출로 인한 폐 세포 산화스트레스 발생과 세포밀착연접 손상 억제 효과)

  • So, Hee-Jung;Chun, Su-Hyun;Lee, Jin-Won;Lee, Kwang-Won
    • Korean Journal of Food Science and Technology
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    • v.53 no.2
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    • pp.165-173
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    • 2021
  • One of the major sources of air pollution is urban particulate matter (UPM), which causes lung diseases involving oxidative stress, inflammation, and cancer. Codonopsis lanceolata (CL) has been used in East Asia as a traditional oriental medicinal ingredient for lung diseases (e.g., asthma and bronchitis). However, the connection between the impact of CL and UPM in the lungs has rarely been investigated. This study aimed to confirm the inhibitory activity of the ethanol extract of CL (ECL) against oxidative stress and disruption of tight cell junctions in human pulmonary epithelial cells after exposure to UPM. As the lung cells were pre-treated with ECL, the UPM-induced increase in cellular reactive oxygen species production suppressed tight junction proteins (e.g., N-cadherin, fibronectin, occludin, zonula occludens-1, and claudin-4). These results suggest that ECL prevents the possible effects of UPM toxicity on the lungs.

Tight junctional inhibition of entry of Toxoplasma gondii into MDCK cells (MDCK세포의 tight junction 형성이 Toxoplusmu gondii의 숙주세포 침투에 미치는 효과)

  • 남호우;윤지혜
    • Parasites, Hosts and Diseases
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    • v.28 no.4
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    • pp.197-206
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    • 1990
  • Various conditions of cultures were performed to investigate the role of tight junctions formed between adjacent MDCK cells on the entry of Toxoplasma. When MDCK cells were cocultured with excess number of Toxoplasma at the seeding density of 1×105, 3×105, and 5×105 cells/ml for 4 days, the number of intracellular parasites decreased rapidly as the host cells reached saturation density, i.e., the formation of tight junctions. When the concentration of calcium in the media (1.8 mM in general) was shifted to $5{\mu}M$ that resulted in the elimination of tight junction, the penetration of Toxoplasma increased about 2-fold(p<0.05) in the saturated culture, while that of non-saturated culture decreased by half. Trypsin-EDTA which was treated to conquer the tight junctions of saturated culture favored the entry of Toxoplasma about 2.5-fold(P<0.05) compared to the non-treated, while that of non- saturated culture decreased to about one fifth. It was suggested that the tight junctions of epithelial cells play a role as a barrier for the entry of Toxoplasma and Toxoplasma penetrate into hoot cells through membrane structure-specific, i.e., certain kind of receptors present on the basolateral rather than apical surface of MDCK cells.

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Investigation of the Effect of Sappan Lignum and Brazilin on Expression of Tight Junction Related-genes in Human Keratinocyte (소목(蘇木)과 그 지표물질인 brazilin이 인간 유래 각질 형성 세포의 tight junction 유전자 발현에 미치는 영향)

  • Cheon, Seong Hye;Choi, Sun Kyung;Cho, Nam Joon;Kim, Kee Kwang;Lee, Woong Hee;Hwang, Hyung Seo;Kim, Kyoon Eon;Han, Hyosang
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.32 no.2
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    • pp.106-112
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    • 2018
  • The aim of this research was to determine the diverse effects of Sappan Lignum extract and brazilin on human keratinocyte HaCaT cells. We confirmed the antioxidant effect of Sappan Lignum extract and brazilin was analyzed by using an ABTS assay, confirming the efficacy of water extraction method. Also, we examined effect of Sappan Lignum extract and brazilin on the cell viability, using the MTS assay in HaCaT cells. mRNA expression levels of tight junction-related genes associated with skin barrier in HaCaT cells were analyzed using quantitative real-time PCR analysis. Sappan Lignum extract increased the cellular activity of HaCaT cells and the expression of the tight junction-related genes claudin 3, claudin 6, and ZO-2. Brazilin displayed the same effects as that of the extract on HaCaT cells activity and tight junction-related genes expression. Furthermore, dispase assay demonstrated altered cell-cell adhesion strength of Sappan Lignum extract or brazilin treated HaCaT cells. Sappan Lignum extract or brazilin might be an useful ingredient in skin-mosturizinng and anti-wrinkle cosmetics, given its effects of altering mRNA expression of tight junction-related genes and enhancing cell-cell adhesion strength of HaCaT cells.

Study on the pressure self-adaptive water-tight junction box in underwater vehicle

  • Huang, Haocai;Ye, Yanying;Leng, Jianxing;Yuan, Zhuoli;Chen, Ying
    • International Journal of Naval Architecture and Ocean Engineering
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    • v.4 no.3
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    • pp.302-312
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    • 2012
  • Underwater vehicles play a very important role in underwater engineering. Water-tight junction box (WJB) is one of the key components in underwater vehicle. This paper puts forward a pressure self-adaptive water-tight junction box (PSAWJB) which improves the reliability of the WJB significantly by solving the sealing and pressure problems in conventional WJB design. By redundancy design method, the pressure self-adaptive equalizer (PSAE) is designed in such a way that it consists of a piston pressure-adaptive compensator (PPAC) and a titanium film pressure-adaptive compensator (TFPAC). According to hydro-mechanical simulations, the operating volume of the PSAE is more than or equal to 11.6 % of the volume of WJB liquid system. Furthermore, the required operating volume of the PSAE also increases as the gas content of oil, hydrostatic pressure or temperature difference increases. The reliability of the PSAWJB is proved by hyperbaric chamber tests.

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.

Characterization of TRAF4 mRNA and Functions related to tight junction in pig (돼지에서 TRAF4 유전자 특성 및 Tight junction 관련 기능 분석)

  • Yun, Jeong-hee;Hwang, In-Sul;Hwang, Seongsoo;Park, Mi-Ryung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.5
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    • pp.216-222
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    • 2020
  • Tumor necrosis factor receptor associated factor 4 (TRAF4) is found to be overexpressed in human breast cancer. It plays a role in cancer metastasis, production of reactive oxygen species, and cell polarity at membranes. The characteristics and functions of TRAF4 in pigs have not yet been identified. As the first step of research, the mRNA sequence of TRAF4 in porcine cells has been determined. To obtain the full-length sequence, rapid amplification of cDNA ends (RACE) has been carried out. Upon cloning, 2,030 bp of nucleotides were found to encode 470 amino acids, and 8 and 12 amino acids were different from those of the human and mouse TRAF4, respectively. The coding region of porcine TRAF4 was shown to be 93% and 90% homologous to human and mouse TRAF4, respectively. qPCR was conducted to determine the relative expression level of TRAF4. TRAF4 expression in pK15 was enhanced by cell-cell contacts. The mRNA levels of CLDN4, OCLN, and TJP1 at 60% and 80% confluency were significantly higher than at 40% confluency. Further, TRAF4 and tight junction-related genes were down-regulated upon treatment with TRAF4 siRNA. Thus, TRAF4 may affect the function of tight junctions in pig.

Activation of Lysosomal Function Ameliorates Amyloid-β-Induced Tight Junction Disruption in the Retinal Pigment Epithelium

  • Dong Hyun Jo;Su Hyun Lee;Minsol Jeon;Chang Sik Cho;Da-Eun Kim;Hyunkyung Kim;Jeong Hun Kim
    • Molecules and Cells
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    • v.46 no.11
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    • pp.675-687
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    • 2023
  • Accumulation of pathogenic amyloid-β disrupts the tight junction of retinal pigment epithelium (RPE), one of its senescence-like structural alterations. In the clearance of amyloid-β, the autophagy-lysosome pathway plays the crucial role. In this context, mammalian target of rapamycin (mTOR) inhibits the process of autophagy and lysosomal degradation, acting as a potential therapeutic target for age-associated disorders. However, efficacy of targeting mTOR to treat age-related macular degeneration remains largely elusive. Here, we validated the therapeutic efficacy of the mTOR inhibitors, Torin and PP242, in clearing amyloid-β by inducing the autophagy-lysosome pathway in a mouse model with pathogenic amyloid-β with tight junction disruption of RPE, which is evident in dry age-related macular degeneration. High concentration of amyloid-β oligomers induced autophagy-lysosome pathway impairment accompanied by the accumulation of p62 and decreased lysosomal activity in RPE cells. However, Torin and PP242 treatment restored the lysosomal activity via activation of LAMP2 and facilitated the clearance of amyloid-β in vitro and in vivo. Furthermore, clearance of amyloid-β by Torin and PP242 ameliorated the tight junction disruption of RPE in vivo. Overall, our findings suggest mTOR inhibition as a new therapeutic strategy for the restoration of tight junctions in age-related macular degeneration.

Effect of Korean Red Ginseng on the Stability of the Tight Junction of Intestinal Epithelial Cells (홍삼에 의한 Caco-2 단세포층 간극의 안정화)

  • Shon, Dong-Hwa;Kim, Mi-Hye;Kim, Young-Chan;Kim, Sung-Soo
    • Korean Journal of Food Science and Technology
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    • v.42 no.3
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    • pp.335-342
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    • 2010
  • Bioactive components involved in the tight junction stabilization of intestinal epithelial cells from Korean red ginseng were studied by analyzing transepithelial electrical resistance (TEER) values of the Caco-2 cell monolayer between the apical and basolateral sides for 96 hr. The treatment with less than $20\;{\mu}g/mL$ of the Korean red ginseng extract to the apical side of Caco-2 cell monolayer gave higher TEER values than the control. However, the treatment with more than $130\;{\mu}g/mL$ of the Korean red ginseng extract drastically decreased the TEER values, and these effects were not due to its cytotoxicity. When fractions of low molecular weight compounds, polysaccharides, proteins, saponins, and polyphenols derived from Korean ginseng were applied to the apical side of the Caco-2 cell monolayer, polyphenols showed high tight junction stabilizing activity and saponins showed low activity, but the others showed no significant activity. These results suggest that Korean red ginseng might be useful for the prevention of food allergy by stabilizing the tight junction of intestinal epithelial cells leading to hindering absorption of food allergens.

Inhibition of NAD(P)H:Quinone Oxidoreductase 1 by Dicumarol Reduces Tight Junction in Human Colonic Epithelial Cells (인간 대장상피세포 밀착연접 형성과정에서 NQO1 저해 효과)

  • Hong, Ji;Zhang, Peng;Yoon, I Na;Kim, Ho
    • Journal of Life Science
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    • v.26 no.5
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    • pp.531-536
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    • 2016
  • We previously showed that NAD(P)H:quinone oxidoreductase 1 (NQO1) knockout (KO) mice exhibited spontaneous inflammation with markedly increased mucosal permeability in the gut, and that NQO1 is functionally associated with regulating tight junctions in the mucosal epithelial cells that govern the mucosal barrier. Here, we confirm the role of NQO1 in the formation of tight junctions by human colonic epithelial cells (HT29). We treated HT29 cells with a chemical inhibitor of NQO1 (dicumarol; 10 μM), and examined the effect on the transepithelial resistance of epithelial cells and the protein expression levels of ZO1 and occludin (two known regulators of tight junctions between gut epithelial cells). The dicumarol-induced inhibition of NQO1 markedly reduced transepithelial resistance (a measure of tight junctions) and decreased the levels of the tested tight junction proteins. In vivo, luminal injection of dicumarol significantly increased mucosal permeability and decreased ZO1 and occludin protein expression levels in mouse guts. However, in contrast to the previous report that the epithelial cells of NQO1 KO mice showed marked down-regulations of the transcripts encoding ZO1 and occludin, these transcript levels were not affected in dicumarol-treated HT29 cells. This result suggests that the NQO1-depedent regulation of tight junction molecules may involve multiple processes, including both transcriptional regulation and protein degradation processes such as those governed by the ubiquitination/proteasomal, and/or lysosomal systems.