Pharmaceutical Activation or Genetic Absence of ClC-2 Alters Tight Junctions During Experimental Colitis

Inflamm Bowel Dis. 2015 Dec;21(12):2747-57. doi: 10.1097/MIB.0000000000000550.

Abstract

Background: We have previously reported that the ClC-2 chloride channel has an important role in regulation of tight junction barrier function during experimental colitis, and the pharmaceutical ClC-2 activator lubiprostone initiates intestinal barrier repair in ischemic-injured intestine. Thus, we hypothesized that pharmaceutical ClC-2 activation would have a protective and therapeutic effect in murine models of colitis, which would be absent in ClC-2 mice.

Methods: We administered lubiprostone to wild-type or ClC-2 mice with dextran sulfate sodium (DSS) or 2, 4, 5-trinitrobenzene sulfonic acid-induced colitis. We determined the severity of colitis and assessed intestinal permeability. Selected tight junction proteins were analyzed by Western blotting and immunofluorescence/confocal microscopy, whereas proliferative and differentiated cells were examined with special staining and immunohistochemistry.

Results: Oral preventive or therapeutic administration of lubiprostone significantly reduced the severity of colitis and reduced intestinal permeability in both DSS and trinitrobenzene sulfonic acid-induced colitis. Preventive treatment with lubiprostone induced significant recovery of the expression and distribution of selected sealing tight junction proteins in mice with DSS-induced colitis. In addition, lubiprostone reduced crypt proliferation and increased the number of differentiated epithelial cells. Alternatively, when lubiprostone was administered to ClC-2 mice, the protective effect against DSS colitis was limited.

Conclusions: This study suggests a central role for ClC-2 in restoration of barrier function and tight junction architecture in experimental murine colitis, which can be therapeutically targeted with lubiprostone.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western
  • CLC-2 Chloride Channels
  • Chloride Channel Agonists / pharmacology*
  • Chloride Channels / deficiency
  • Chloride Channels / drug effects*
  • Colitis / chemically induced
  • Colitis / drug therapy*
  • Colitis / genetics
  • Dextran Sulfate
  • Fluorescent Antibody Technique
  • Intestinal Mucosa / metabolism
  • Intestines / drug effects
  • Lubiprostone / pharmacology*
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Confocal
  • Permeability / drug effects
  • Tight Junction Proteins / metabolism
  • Tight Junctions / drug effects*
  • Tight Junctions / metabolism
  • Trinitrobenzenes

Substances

  • CLC-2 Chloride Channels
  • Chloride Channel Agonists
  • Chloride Channels
  • Tight Junction Proteins
  • Trinitrobenzenes
  • Lubiprostone
  • Dextran Sulfate