Phosphodiesterase inhibition attenuates alterations to the tight junction proteins occludin and ZO-1 in immunostimulated Caco-2 intestinal monolayers

Todd W. Costantini, Jessica Deree, William Loomis, James G. Putnam, Sung Hyuk Choi, Andrew Baird, Brian P. Eliceiri, Vishal Bansal, Raul Coimbra

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Aims: Under normal conditions, the intestinal mucosa acts as a local barrier to prevent the influx of luminal contents. The intestinal epithelial tight junction is comprised of several membrane associated proteins, including zonula occludens-1 (ZO-1) and occludin. Disruption of this barrier can lead to the production of pro-inflammatory mediators and ultimately multiple organ failure. We have previously shown that Pentoxifylline (PTX) decreases histologic gut injury and pro-inflammatory mediator synthesis. We hypothesize that PTX prevents the breakdown of ZO-1 and occludin in an in vitro model of immunostimulated intestinal cell monolayers. Main methods: Caco-2 human enterocytes were grown as confluent monolayers and incubated under control conditions, or with PTX (2 mM), Cytomix (TNF-α, IFN-γ, IL-1), or Cytomix + PTX for 24 h. Occludin and ZO-1 protein levels were analyzed by Western blot. Confocal microscopy was used to assess the cytoplasmic localization of ZO-1 and occludin. Key findings: Cytomix stimulation of Caco-2 cells resulted in a 50% decrease in both occludin and ZO-1 protein. Treatment with Cytomix + PTX restored both occludin and ZO-1 protein to control levels. Confocal microscopy images show that Cytomix caused an irregular, undulating appearance of ZO-1 and occludin at the cell junctions. Treatment with PTX prevented the Cytomix-induced changes in ZO-1 and occludin localization. Significance: Treatment with PTX decreases the pro-inflammatory cytokine induced changes in the intestinal tight junction proteins occludin and ZO-1. Pentoxifylline may be a useful adjunct in the treatment of sepsis and shock by attenuating intestinal barrier breakdown.

Original languageEnglish
Pages (from-to)18-22
Number of pages5
JournalLife Sciences
Volume84
Issue number1-2
DOIs
Publication statusPublished - 2009 Jan 2
Externally publishedYes

Fingerprint

Occludin
Tight Junction Proteins
Tight Junctions
Pentoxifylline
Phosphoric Diester Hydrolases
Monolayers
Zonula Occludens-1 Protein
Confocal microscopy
Confocal Microscopy
Intercellular Junctions
Caco-2 Cells
Multiple Organ Failure
Enterocytes
Level control
Intestinal Mucosa
Interleukin-1
Shock
Sepsis
Membrane Proteins
Western Blotting

Keywords

  • Caco-2 cells
  • Cytokines
  • Inflammation
  • Intestinal permeability
  • Occludin
  • Pentoxifylline
  • Phosphodiesterase inhibition
  • Shock
  • Tight junction
  • ZO-1

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)

Cite this

Phosphodiesterase inhibition attenuates alterations to the tight junction proteins occludin and ZO-1 in immunostimulated Caco-2 intestinal monolayers. / Costantini, Todd W.; Deree, Jessica; Loomis, William; Putnam, James G.; Choi, Sung Hyuk; Baird, Andrew; Eliceiri, Brian P.; Bansal, Vishal; Coimbra, Raul.

In: Life Sciences, Vol. 84, No. 1-2, 02.01.2009, p. 18-22.

Research output: Contribution to journalArticle

Costantini, Todd W. ; Deree, Jessica ; Loomis, William ; Putnam, James G. ; Choi, Sung Hyuk ; Baird, Andrew ; Eliceiri, Brian P. ; Bansal, Vishal ; Coimbra, Raul. / Phosphodiesterase inhibition attenuates alterations to the tight junction proteins occludin and ZO-1 in immunostimulated Caco-2 intestinal monolayers. In: Life Sciences. 2009 ; Vol. 84, No. 1-2. pp. 18-22.
@article{6d7e3dc8079d4d4181fc11244086fc16,
title = "Phosphodiesterase inhibition attenuates alterations to the tight junction proteins occludin and ZO-1 in immunostimulated Caco-2 intestinal monolayers",
abstract = "Aims: Under normal conditions, the intestinal mucosa acts as a local barrier to prevent the influx of luminal contents. The intestinal epithelial tight junction is comprised of several membrane associated proteins, including zonula occludens-1 (ZO-1) and occludin. Disruption of this barrier can lead to the production of pro-inflammatory mediators and ultimately multiple organ failure. We have previously shown that Pentoxifylline (PTX) decreases histologic gut injury and pro-inflammatory mediator synthesis. We hypothesize that PTX prevents the breakdown of ZO-1 and occludin in an in vitro model of immunostimulated intestinal cell monolayers. Main methods: Caco-2 human enterocytes were grown as confluent monolayers and incubated under control conditions, or with PTX (2 mM), Cytomix (TNF-α, IFN-γ, IL-1), or Cytomix + PTX for 24 h. Occludin and ZO-1 protein levels were analyzed by Western blot. Confocal microscopy was used to assess the cytoplasmic localization of ZO-1 and occludin. Key findings: Cytomix stimulation of Caco-2 cells resulted in a 50{\%} decrease in both occludin and ZO-1 protein. Treatment with Cytomix + PTX restored both occludin and ZO-1 protein to control levels. Confocal microscopy images show that Cytomix caused an irregular, undulating appearance of ZO-1 and occludin at the cell junctions. Treatment with PTX prevented the Cytomix-induced changes in ZO-1 and occludin localization. Significance: Treatment with PTX decreases the pro-inflammatory cytokine induced changes in the intestinal tight junction proteins occludin and ZO-1. Pentoxifylline may be a useful adjunct in the treatment of sepsis and shock by attenuating intestinal barrier breakdown.",
keywords = "Caco-2 cells, Cytokines, Inflammation, Intestinal permeability, Occludin, Pentoxifylline, Phosphodiesterase inhibition, Shock, Tight junction, ZO-1",
author = "Costantini, {Todd W.} and Jessica Deree and William Loomis and Putnam, {James G.} and Choi, {Sung Hyuk} and Andrew Baird and Eliceiri, {Brian P.} and Vishal Bansal and Raul Coimbra",
year = "2009",
month = "1",
day = "2",
doi = "10.1016/j.lfs.2008.10.007",
language = "English",
volume = "84",
pages = "18--22",
journal = "Life Sciences",
issn = "0024-3205",
publisher = "Elsevier Inc.",
number = "1-2",

}

TY - JOUR

T1 - Phosphodiesterase inhibition attenuates alterations to the tight junction proteins occludin and ZO-1 in immunostimulated Caco-2 intestinal monolayers

AU - Costantini, Todd W.

AU - Deree, Jessica

AU - Loomis, William

AU - Putnam, James G.

AU - Choi, Sung Hyuk

AU - Baird, Andrew

AU - Eliceiri, Brian P.

AU - Bansal, Vishal

AU - Coimbra, Raul

PY - 2009/1/2

Y1 - 2009/1/2

N2 - Aims: Under normal conditions, the intestinal mucosa acts as a local barrier to prevent the influx of luminal contents. The intestinal epithelial tight junction is comprised of several membrane associated proteins, including zonula occludens-1 (ZO-1) and occludin. Disruption of this barrier can lead to the production of pro-inflammatory mediators and ultimately multiple organ failure. We have previously shown that Pentoxifylline (PTX) decreases histologic gut injury and pro-inflammatory mediator synthesis. We hypothesize that PTX prevents the breakdown of ZO-1 and occludin in an in vitro model of immunostimulated intestinal cell monolayers. Main methods: Caco-2 human enterocytes were grown as confluent monolayers and incubated under control conditions, or with PTX (2 mM), Cytomix (TNF-α, IFN-γ, IL-1), or Cytomix + PTX for 24 h. Occludin and ZO-1 protein levels were analyzed by Western blot. Confocal microscopy was used to assess the cytoplasmic localization of ZO-1 and occludin. Key findings: Cytomix stimulation of Caco-2 cells resulted in a 50% decrease in both occludin and ZO-1 protein. Treatment with Cytomix + PTX restored both occludin and ZO-1 protein to control levels. Confocal microscopy images show that Cytomix caused an irregular, undulating appearance of ZO-1 and occludin at the cell junctions. Treatment with PTX prevented the Cytomix-induced changes in ZO-1 and occludin localization. Significance: Treatment with PTX decreases the pro-inflammatory cytokine induced changes in the intestinal tight junction proteins occludin and ZO-1. Pentoxifylline may be a useful adjunct in the treatment of sepsis and shock by attenuating intestinal barrier breakdown.

AB - Aims: Under normal conditions, the intestinal mucosa acts as a local barrier to prevent the influx of luminal contents. The intestinal epithelial tight junction is comprised of several membrane associated proteins, including zonula occludens-1 (ZO-1) and occludin. Disruption of this barrier can lead to the production of pro-inflammatory mediators and ultimately multiple organ failure. We have previously shown that Pentoxifylline (PTX) decreases histologic gut injury and pro-inflammatory mediator synthesis. We hypothesize that PTX prevents the breakdown of ZO-1 and occludin in an in vitro model of immunostimulated intestinal cell monolayers. Main methods: Caco-2 human enterocytes were grown as confluent monolayers and incubated under control conditions, or with PTX (2 mM), Cytomix (TNF-α, IFN-γ, IL-1), or Cytomix + PTX for 24 h. Occludin and ZO-1 protein levels were analyzed by Western blot. Confocal microscopy was used to assess the cytoplasmic localization of ZO-1 and occludin. Key findings: Cytomix stimulation of Caco-2 cells resulted in a 50% decrease in both occludin and ZO-1 protein. Treatment with Cytomix + PTX restored both occludin and ZO-1 protein to control levels. Confocal microscopy images show that Cytomix caused an irregular, undulating appearance of ZO-1 and occludin at the cell junctions. Treatment with PTX prevented the Cytomix-induced changes in ZO-1 and occludin localization. Significance: Treatment with PTX decreases the pro-inflammatory cytokine induced changes in the intestinal tight junction proteins occludin and ZO-1. Pentoxifylline may be a useful adjunct in the treatment of sepsis and shock by attenuating intestinal barrier breakdown.

KW - Caco-2 cells

KW - Cytokines

KW - Inflammation

KW - Intestinal permeability

KW - Occludin

KW - Pentoxifylline

KW - Phosphodiesterase inhibition

KW - Shock

KW - Tight junction

KW - ZO-1

UR - http://www.scopus.com/inward/record.url?scp=57749185476&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=57749185476&partnerID=8YFLogxK

U2 - 10.1016/j.lfs.2008.10.007

DO - 10.1016/j.lfs.2008.10.007

M3 - Article

C2 - 18992758

AN - SCOPUS:57749185476

VL - 84

SP - 18

EP - 22

JO - Life Sciences

JF - Life Sciences

SN - 0024-3205

IS - 1-2

ER -