Vasoactive intestinal peptide ameliorates intestinal barrier disruption associated with Citrobacter rodentium-induced colitis.
Attaching and effacing bacterial pathogens attach to the apical surface of epithelial cells and disrupt epithelial barrier function, increasing permeability and allowing luminal contents access to the underlying milieu. Previous in vitro studies demonstrated that the neuropeptide vasoactive intestinal peptide (VIP) regulates epithelial paracellular permeability, and the high concentrations and close proximity of VIP-containing nerve fibers to intestinal epithelial cells would support such a function in vivo. The aim of this study was to examine whether VIP treatment modulated Citrobacter rodentium-induced disruption of intestinal barrier integrity and to identify potential mechanisms of action. Administration of VIP had no effect on bacterial attachment although histopathological scoring demonstrated a VIP-induced amelioration of colitis-induced epithelial damage compared with controls. VIP treatment prevented the infection-induced increase in mannitol flux a measure of paracellular permeability, resulting in levels similar to control mice, and immunohistochemical studies demonstrated that VIP prevented the translocation of tight junction proteins: zonula occludens-1, occludin, and claudin-3. Enteropathogenic Escherichia coli (EPEC) infection of Caco-2 monolayers confirmed a protective role for VIP on epithelial barrier function. VIP prevented EPEC-induced increase in long myosin light chain kinase (MLCK) expression and myosin light chain phosphorylation (p-MLC). Furthermore, MLCK inhibition significantly attenuated bacterial-induced epithelial damage both in vivo and in vitro. In conclusion, our results indicate that VIP protects the colonic epithelial barrier by minimizing bacterial-induced redistribution of tight junction proteins in part through actions on MLCK and MLC phosphorylation.
Conlin
V S
VS
Division of Gastroenterology, Child and Family Research Institute, BC Children's Hospital, Vancouver, BC, Canada.
Wu
X
X
Nguyen
C
C
Dai
C
C
Vallance
B A
BA
Buchan
A M J
AM
Boyer
L
L
Jacobson
K
K
D000818:Animals; D000893:Anti-Inflammatory Agents / Q000008:administration & dosage*; D001381:Azepines / Q000494:pharmacology; D001422:Bacterial Adhesion; D018988:Bacterial Translocation / Q000187:drug effects*; D018938:Caco-2 Cells; D044082:Citrobacter rodentium / Q000472:pathogenicity*; D062465:Claudin-3; D003092:Colitis / Q000378:metabolism / Q000382:microbiology / Q000473:pathology / Q000517:prevention & control*; D003106:Colon / Q000187:drug effects* / Q000378:metabolism / Q000382:microbiology / Q000473:pathology; D004195:Disease Models, Animal; D004756:Enterobacteriaceae Infections / Q000150:complications / Q000188:drug therapy* / Q000378:metabolism / Q000382:microbiology; D006801:Humans; D007274:Injections, Intraperitoneal; D007413:Intestinal Mucosa / Q000187:drug effects* / Q000378:metabolism / Q000382:microbiology; D008353:Mannitol / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018994:Myosin Light Chains / Q000378:metabolism; D009219:Myosin-Light-Chain Kinase / Q000037:antagonists & inhibitors / Q000378:metabolism; D009281:Naphthalenes / Q000494:pharmacology; D062793:Occludin; D010539:Permeability; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D019108:Tight Junctions / Q000187:drug effects / Q000378:metabolism / Q000382:microbiology; D013997:Time Factors; D014660:Vasoactive Intestinal Peptide / Q000008:administration & dosage*; D062826:Zonula Occludens-1 Protein
D000893:Anti-Inflammatory Agents; D001381:Azepines; C568122:CLDN3 protein, human; D062465:Claudin-3; C568123:Cldn3 protein, mouse; D008565:Membrane Proteins; D018994:Myosin Light Chains; D009281:Naphthalenes; C568985:OCLN protein, human; D062793:Occludin; C568986:Ocln protein, mouse; D010750:Phosphoproteins; D047428:Protein Kinase Inhibitors; C569042:TJP1 protein, human; C569043:Tjp1 protein, mouse; D062826:Zonula Occludens-1 Protein; C070571:ML 7; D014660:Vasoactive Intestinal Peptide; D008353:Mannitol; D009219:Myosin-Light-Chain Kinase