• Title/Summary/Keyword: transepithelial electrical resistance

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Assessment of Reproductive Health Risk of Polychlorinated Biphenyls by Monitoring the Expression of Claudius and Transepithelial Electrical Resistance in Mouse Sertoli Cells

  • Gye, Myung-Chan;Seiichiroh Ohsako;Lee, Ho-Joon
    • Journal of Microbiology and Biotechnology
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    • v.13 no.4
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    • pp.495-500
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    • 2003
  • Tight junctions (TJ) between adjacent Sertoli cells in testis are important for the formation of the blood testis barrier (BTB). In an effort to verify the reproductive health risk of endocrine-active chemicals (EACs), changes in the transepithelial electrical resistance (TER) and the expression of TJ genes were examined by co-planar polychlorinated biphenyl (PCB) treatment in cultured mouse Sertoli cells. Although the increase in TER of Sertoli cells was accelerated by 10 nM co-planar PCB, it was downregulated by 100 nM co-planar PCB. The expression of claudin-1 was downregulated by co-planar PCB in a concentration-dependent manner. On the contrary, the expression of claudin-1 was increased in the Sertoli cells by 10 nM co-planar PCB treatment. These results suggest that the structure and function of TJ may be targeted by co-planar PCB in Sertoli cells. Assessment of the structure and function of TJ in Sertoli cells might be useful for screening the reproductive health risk of EACs.

Development of an Escherichia coli Biofilm Model on Transwell®

  • Kim, Bok Yung;Thyiam, General;Kang, Ji-Eun;Lee, Seung-Hwan;Park, Sang-Hee;Kim, Jung-Sun;Abraham, Marion
    • Korean Journal of Clinical Laboratory Science
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    • v.44 no.3
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    • pp.112-117
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    • 2012
  • Escherichia coli biofilm, reported to be produced in the human intestine causing a significant health risk, was successfully grown on transwell$^{(R)}$. This biofilm layer was identified by crystal violet staining and prepared for the in vitro E. coli biofilm system which can be used to screen for inhibitors. The biofilm formation did not show a change in transepithelial electrical resistance values. Furthermore, rhodamine 123 staining showed that the dye did not pass through the membrane once biofilm was formed.

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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.

The Effects of Glucagon-like Peptide-2 on the Tight Junction and Barrier Function in IPEC-J2 Cells through Phosphatidylinositol 3-kinase-Protein Kinase B-Mammalian Target of Rapamycin Signaling Pathway

  • Yu, Changsong;Jia, Gang;Deng, Qiuhong;Zhao, Hua;Chen, Xiaoling;Liu, Guangmang;Wang, Kangning
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.5
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    • pp.731-738
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    • 2016
  • Glucagon-like peptide-2 (GLP-2) is important for intestinal barrier function and regulation of tight junction (TJ) proteins, but the intracellular mechanisms of action remain undefined. The purpose of this research was to determine the protective effect of GLP-2 mediated TJ and transepithelial electrical resistance (TER) in lipopolysaccharide (LPS) stressed IPEC-J2 cells and to test the hypothesis that GLP-2 regulate TJ and TER through the phosphatidylinositol 3-kinase (PI3K)-protein kinase B (Akt)-mammalian target of rapamycin (mTOR) signaling pathway in IPEC-J2 cells. Wortmannin and LY294002 are specific inhibitors of PI3K. The results showed that $100{\mu}g/mL$ LPS stress decreased TER and TJ proteins occludin, claudin-1 and zonula occludens protein 1 (ZO-1) mRNA, proteins expressions (p<0.01) respectively. GLP-2 (100 nmol/L) promote TER and TJ proteins occludin, claudin-1, and zo-1 mRNA, proteins expressions in LPS stressed and normal IPEC-J2 cells (p<0.01) respectively. In normal cells, both wortmannin and LY294002, PI3K inhibitors, prevented the mRNA and protein expressions of Akt and mTOR increase induced by GLP-2 (p<0.01) following with the significant decreasing of occludin, claudin-1, ZO-1 mRNA and proteins expressions and TER (p<0.01). In conclusion, these results indicated that GLP-2 can promote TJ's expression and TER in LPS stressed and normal IPEC-J2 cells and GLP-2 could regulate TJ and TER through the PI3K/Akt/mTOR pathway.

Inhibition of Cell Invasion by Ethyl Alcohol Extracts of Hizikia fusiforme in AGS Human Gastric Adenocarcinoma Cells (AGS 인체 위암세포에서 톳 에탄올 추출물에 의한 침윤성 저해)

  • Choi, Yung-Hyun
    • Journal of Life Science
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    • v.20 no.12
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    • pp.1784-1791
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    • 2010
  • In this study, we investigated the effects of ethyl alcohol extracts of Hizikia fusiforme (EHF) on the correlation between tightening of tight junctions (TJs) and anti-invasive activity in human gastric adenocarcinoma AGS cells. Inhibitory effects of EHF on cell proliferation, motility, and invasiveness were found to be associated with increased tightness of the TJs, which was demonstrated by an increase in transepithelial electrical resistance. Activities of matrix metalloprotease (MMP)-2 and -9 in AGS cells were dose-dependently inhibited by treatment with EHF, and this was also correlated with a decrease in expression of their mRNA and proteins; however, tissue inhibitor of metalloproteinase (TIMP)-1 and -2 mRNA levels were increased. Additionally, immunoblotting results indicated that EHF repressed the levels of claudin proteins (claudin-1, -3, and -4), major components of TJs that play key roles in control and selectivity of paracellular transport. Furthermore, EHF decreased expression of insulin such as growth factor-1 receptor proteins, while concurrently increasing that of thrombospondin-1 and E-cadherin. In conclusion, these results suggest that EHF treatment may inhibit tumor cell motility and invasion, and therefore act as a dietary source to decrease the risk of cancer metastasis.

Effects of BuOH Extract of the Root of Aralia elata as an Absorption Enhancer on the Transport of Chondroitin Sulfate and Its Digestion Products In Vitro and In Vivo

  • Sim, Joon-Soo;Li, Da-Wei;Cho, Hai-Lim;Cho, So-Yean;Jeong, Choon-Sik;Lee, Eun-Bang;Kim, Yeong-Shik
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.415.2-415.2
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    • 2002
  • We investigated the absorption enhancing effect of BuOH extract of the root of Aralia elata (BERAE) in Caco-2 cell monolayers and rats. At the concentration of both 0.04% and 0.08% (w/v). BERAE decreased the transepithelial electrical resistance (TEER) values and increased the permeability of intact chondroitin sulfate (CS) and its digestion products as hydrophilic macromolecules in a dose dependent manner. We also evaluated the cytotoxicity of BERAE for the determination of a proper concentration as an absorption enhancer. (omitted)

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Investigation of transport of PEGylated salmon calcitonin through caco-2 cell monolayers

  • Oh, Seung-Huyn;Youn, Yu-Seok;Lee, Jeong-Eun;Park, Yun-Sang;Lee, Kang-Choon
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.234.3-235
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    • 2003
  • The aim of this study is to evaluate the permeability of PEG-conjugated salmon calcitonin (sCT) across monolayers of Caco-2 cells that represent a model of the intestinal barrier. Caco-2 cells were grown to confluency on a permeable polycarbonate membrane to permit transport through it. Permeability experiments were performed with native-sCT and PEG-conjugated sCT (PEG M.W. 2000) at various concentrations (5uM, 10uM, 25uM, 50uM, 100uM) in the apical to basolateral direction. The barrier properties were assessed by detecting transport of markder molecules ($^3$H-mannitol) and by measuring transepithelial electrical resistance (TEER). (omitted)

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Anti-invasive Activity of Human Breast Carcinoma Cells by Genistein through Modulation of Tight Junction Function (인체유방암세포의 tight junction 기능 조절을 통한 genistein의 암세포 침윤 억제 효과)

  • Kim, Sung-Ok;Jeang, Yang-Kee;Choi, Yung-Hyun
    • Journal of Life Science
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    • v.19 no.9
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    • pp.1200-1208
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    • 2009
  • Tight junctions (TJs) that act as paracellular permeability barriers play an essential role in regulating the diffusion of fluid, electrolytes and macromolecules through the paracellular pathway. In this study, we investigated the correlation between the tightening of TJs, permeability and the invasive activity of genistein - a bioactive isoflavone of soybeans - in human breast carcinoma MCF-7 and MDA-MB-231 cells. The inhibitory effects of genistein on cell proliferation, motility and invasiveness were found to be associated with the increased tightness of the TJs, which was demonstrated by an increase in transepithelial electrical resistance and a decrease in paracellular permeability. Additionally, the immunoblotting results indicated that genistein repressed the levels of the proteins that comprise the major components of TJ, claudin-3 and claudin-4, which play a key role in the control and selectivity of paracellular transport. Furthermore, genistein decreased the metastasis-related gene expressions of insulin like growth factor-1 receptor and snail, while concurrently increasing that of thrombospondin-1 and E-cadherin. In addition, we demonstrated that claudins play an important role in the anti-motility and invasiveness of genistein using claudin-3 small interfering RNA. Taken together, our results indicate a possible role for genistein as an inhibitor of cancer cell invasion through the tightening of TJs, which may counteract the up-regulation of claudins. In addition, our results indicate that this may be beneficial for the inhibition of tumor metastasis.

Effect of ginger and cinnamon extract mixtures on the growth of intestinal bacteria and intestinal inflammation (생강계피 복합물이 장내 유익균 증식 및 염증조절 기능에 미치는 영향)

  • Kim, Min Ju;Kim, Min Seo;Kang, Sung Tae;Kim, Ji Yeon
    • Journal of Applied Biological Chemistry
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    • v.60 no.4
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    • pp.321-326
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    • 2017
  • We aimed to assess the potential growth-promoting effects of ginger and cinnamon mixtures (GCM) on intestinal bacteria and their anti-inflammatory effects in a cellular model of intestinal inflammation. Bifidobacterium longum, Lactobacillus sp., and Lactobacillus acidophilus served as intestinal bacteria. Further, in the inflammatory co-culture model, Caco-2 cells co-cultured with RAW264.7 cells were treated with GCM before the addition of lipopolysaccharide (LPS) to induce inflammation in RAW264.7 cells. Addition of GCM to modified Eggerth Gagnon media at a ginger:cinnamon ratio of 1:5 increased the growth of B. longum, Lactobacillus sp., and L. acidophilus compared to that of the control. In a cellular model, compared to LPS-treated groups, GCM-treated groups maintained high transepithelial electrical resistance at ginger:cinnamon ratios of 1:1, 1:3, 1:5, and 1:7 and decreased the tight junction permeability at 3:1, 1:1, 1:3, and 1:5 ratios, similar to that shown by the control groups. In addition, GCM-treated groups showed decreased levels of nitrite at 1:1, 1:5, and 1:7 ginger:cinnamon ratios. Based on these results, it can be concluded that among the various combinations of GCM, the ginger:cinnamon ratio of 1:5 is the optimal composite ratio that shows positive effects on the intestinal beneficial bacteria and in anti-inflammation.

Inhibition of Cell Invasion by Indole-3-Carbinol in OVCAR-3 Human Ovarian Cancer Cells (Indole-3-carbinol에 의한 OVCAR-3 인체 난소암세포의 침윤 억제)

  • Choi, Yung-Hyun;Kim, Sung-Ok
    • Journal of Life Science
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    • v.21 no.7
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    • pp.923-931
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    • 2011
  • In the present study, we investigated the effect of indole-3-carbinol (I3C), a natural compound present in vegetables, on the cell migration and invasion of OVCAR-3 ovarian cancer cells. Our results indicated that I3C inhibited the proliferation of OVCAR-3 cells, a process which was associated with inhibition of cell motility as determined by wound healing experiments and cell invasion studies. I3C treatment increased the tightness of the tight junctions (TJs), which was demonstrated by an increase in transepithelial electrical resistance and a decrease in paracellular permeability. The RT-PCR and immunoblotting results indicated that I3C repressed the levels of claudin-3 as well as claudin-4, proteins that comprise a major part of TJs and play a key role in the control and selectivity of paracellular transport. Furthermore, the activities of matrix metalloproteinase (MMP)-2 and MMP-9 were also decreased by treatment with I3C, which was connected with the down-regulation of their mRNAs and protein expression. The results suggest that I3C may be expected to inhibit cancer cell metastasis and invasion by restoring TJs and decreasing MMP activity in ovarian cancer cell line OVCAR-3.