Conditioned medium from LS 174T goblet cells treated with oxyresveratrol strengthens tight junctions in Caco-2 cells

Dahyun Hwang, Hyun A. Jo, Seonwook Hwang, Jeong Keun Kim, In Ho Kim, Young Hee Lim

Research output: Contribution to journalArticle

Abstract

Background: Strengthening of intestinal tight junctions provides an effective barrier from the external environment. Goblet cell-derived trefoil factor 3 (TFF3) increases transepithelial resistance by upregulating the expression of tight junction proteins. Oxyresveratrol (OXY) is a hydroxyl-substituted stilbene found in the roots, leaves, stems, and fruit of many plants and known to have various biological activities. In this study, we investigated the strengthening effect of OXY on intestinal tight junctions through stimulation of TFF production in goblet cells. Methods: We prepared conditioned medium from LS 174T goblet cells treated with OXY (GCO-CM) and investigated the effect of GCO-CM on strengthening tight junctions of Caco-2 cells. The mRNA and protein expression levels of major tight junction components (claudin-1, occludin, and ZO-1) were measured by quantitative real-time PCR and western blotting, respectively. Transepithelial electric resistance (TEER) was measured using an ohm/V meter. Monolayer permeability was evaluated by paracellular transport of fluorescein isothiocyanate-dextran. Results: OXY showed a strong antioxidant activity. It significantly increased the expression level of TFF3 in LS 174T goblet cells. GCO-CM prepared by treatment with 2.5, 5, and 10. μg/ml OXY did not show cytotoxicity in Caco-2 cells. GCO-CM increased the mRNA and protein expression levels of claudin-1, occludin, and ZO-1. It also significantly increased tight junction integrity and reduced permeability in a dose-dependent manner. Conclusion: OXY stimulates the expression of TFF3 in goblet cells, which might increase the integrity of the intestinal tight junction barrier.

Original languageEnglish
JournalBiomedicine and Pharmacotherapy
DOIs
Publication statusAccepted/In press - 2016 Aug 23

Fingerprint

Goblet Cells
Caco-2 Cells
Tight Junctions
Conditioned Culture Medium
Claudin-1
Occludin
Permeability
Tight Junction Proteins
Stilbenes
Messenger RNA
Electric Impedance
Hydroxyl Radical
puag-haad
Real-Time Polymerase Chain Reaction
Fruit
Proteins
Antioxidants
Western Blotting
Trefoil Factor-3

Keywords

  • Goblet cell
  • Oxyresveratrol
  • TEER
  • TFF3
  • Tight junction

ASJC Scopus subject areas

  • Pharmacology

Cite this

Conditioned medium from LS 174T goblet cells treated with oxyresveratrol strengthens tight junctions in Caco-2 cells. / Hwang, Dahyun; Jo, Hyun A.; Hwang, Seonwook; Kim, Jeong Keun; Kim, In Ho; Lim, Young Hee.

In: Biomedicine and Pharmacotherapy, 23.08.2016.

Research output: Contribution to journalArticle

@article{a5c95c1476b74f9d8019c9ea387091c5,
title = "Conditioned medium from LS 174T goblet cells treated with oxyresveratrol strengthens tight junctions in Caco-2 cells",
abstract = "Background: Strengthening of intestinal tight junctions provides an effective barrier from the external environment. Goblet cell-derived trefoil factor 3 (TFF3) increases transepithelial resistance by upregulating the expression of tight junction proteins. Oxyresveratrol (OXY) is a hydroxyl-substituted stilbene found in the roots, leaves, stems, and fruit of many plants and known to have various biological activities. In this study, we investigated the strengthening effect of OXY on intestinal tight junctions through stimulation of TFF production in goblet cells. Methods: We prepared conditioned medium from LS 174T goblet cells treated with OXY (GCO-CM) and investigated the effect of GCO-CM on strengthening tight junctions of Caco-2 cells. The mRNA and protein expression levels of major tight junction components (claudin-1, occludin, and ZO-1) were measured by quantitative real-time PCR and western blotting, respectively. Transepithelial electric resistance (TEER) was measured using an ohm/V meter. Monolayer permeability was evaluated by paracellular transport of fluorescein isothiocyanate-dextran. Results: OXY showed a strong antioxidant activity. It significantly increased the expression level of TFF3 in LS 174T goblet cells. GCO-CM prepared by treatment with 2.5, 5, and 10. μg/ml OXY did not show cytotoxicity in Caco-2 cells. GCO-CM increased the mRNA and protein expression levels of claudin-1, occludin, and ZO-1. It also significantly increased tight junction integrity and reduced permeability in a dose-dependent manner. Conclusion: OXY stimulates the expression of TFF3 in goblet cells, which might increase the integrity of the intestinal tight junction barrier.",
keywords = "Goblet cell, Oxyresveratrol, TEER, TFF3, Tight junction",
author = "Dahyun Hwang and Jo, {Hyun A.} and Seonwook Hwang and Kim, {Jeong Keun} and Kim, {In Ho} and Lim, {Young Hee}",
year = "2016",
month = "8",
day = "23",
doi = "10.1016/j.biopha.2016.11.022",
language = "English",
journal = "Biomedicine and Pharmacotherapy",
issn = "0753-3322",
publisher = "Elsevier Masson",

}

TY - JOUR

T1 - Conditioned medium from LS 174T goblet cells treated with oxyresveratrol strengthens tight junctions in Caco-2 cells

AU - Hwang, Dahyun

AU - Jo, Hyun A.

AU - Hwang, Seonwook

AU - Kim, Jeong Keun

AU - Kim, In Ho

AU - Lim, Young Hee

PY - 2016/8/23

Y1 - 2016/8/23

N2 - Background: Strengthening of intestinal tight junctions provides an effective barrier from the external environment. Goblet cell-derived trefoil factor 3 (TFF3) increases transepithelial resistance by upregulating the expression of tight junction proteins. Oxyresveratrol (OXY) is a hydroxyl-substituted stilbene found in the roots, leaves, stems, and fruit of many plants and known to have various biological activities. In this study, we investigated the strengthening effect of OXY on intestinal tight junctions through stimulation of TFF production in goblet cells. Methods: We prepared conditioned medium from LS 174T goblet cells treated with OXY (GCO-CM) and investigated the effect of GCO-CM on strengthening tight junctions of Caco-2 cells. The mRNA and protein expression levels of major tight junction components (claudin-1, occludin, and ZO-1) were measured by quantitative real-time PCR and western blotting, respectively. Transepithelial electric resistance (TEER) was measured using an ohm/V meter. Monolayer permeability was evaluated by paracellular transport of fluorescein isothiocyanate-dextran. Results: OXY showed a strong antioxidant activity. It significantly increased the expression level of TFF3 in LS 174T goblet cells. GCO-CM prepared by treatment with 2.5, 5, and 10. μg/ml OXY did not show cytotoxicity in Caco-2 cells. GCO-CM increased the mRNA and protein expression levels of claudin-1, occludin, and ZO-1. It also significantly increased tight junction integrity and reduced permeability in a dose-dependent manner. Conclusion: OXY stimulates the expression of TFF3 in goblet cells, which might increase the integrity of the intestinal tight junction barrier.

AB - Background: Strengthening of intestinal tight junctions provides an effective barrier from the external environment. Goblet cell-derived trefoil factor 3 (TFF3) increases transepithelial resistance by upregulating the expression of tight junction proteins. Oxyresveratrol (OXY) is a hydroxyl-substituted stilbene found in the roots, leaves, stems, and fruit of many plants and known to have various biological activities. In this study, we investigated the strengthening effect of OXY on intestinal tight junctions through stimulation of TFF production in goblet cells. Methods: We prepared conditioned medium from LS 174T goblet cells treated with OXY (GCO-CM) and investigated the effect of GCO-CM on strengthening tight junctions of Caco-2 cells. The mRNA and protein expression levels of major tight junction components (claudin-1, occludin, and ZO-1) were measured by quantitative real-time PCR and western blotting, respectively. Transepithelial electric resistance (TEER) was measured using an ohm/V meter. Monolayer permeability was evaluated by paracellular transport of fluorescein isothiocyanate-dextran. Results: OXY showed a strong antioxidant activity. It significantly increased the expression level of TFF3 in LS 174T goblet cells. GCO-CM prepared by treatment with 2.5, 5, and 10. μg/ml OXY did not show cytotoxicity in Caco-2 cells. GCO-CM increased the mRNA and protein expression levels of claudin-1, occludin, and ZO-1. It also significantly increased tight junction integrity and reduced permeability in a dose-dependent manner. Conclusion: OXY stimulates the expression of TFF3 in goblet cells, which might increase the integrity of the intestinal tight junction barrier.

KW - Goblet cell

KW - Oxyresveratrol

KW - TEER

KW - TFF3

KW - Tight junction

UR - http://www.scopus.com/inward/record.url?scp=85007008347&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85007008347&partnerID=8YFLogxK

U2 - 10.1016/j.biopha.2016.11.022

DO - 10.1016/j.biopha.2016.11.022

M3 - Article

JO - Biomedicine and Pharmacotherapy

JF - Biomedicine and Pharmacotherapy

SN - 0753-3322

ER -