Pharmaceutical Research

, Volume 25, Issue 6, pp 1377–1386 | Cite as

Myosin Light Chain Kinase Inhibition: Correction of Increased Intestinal Epithelial Permeability In Vitro

  • Linda M. Feighery
  • Sean W. Cochrane
  • Teresa Quinn
  • Alan W. Baird
  • Daniel O’Toole
  • Sian-Eleri Owens
  • Diarmuid O’Donoghue
  • Randall J. Mrsny
  • David J. Brayden
Research Paper

Abstract

Purpose

To examine whether myosin light chain kinase (MLCK) inhibitors can reduce intestinal epithelial permeability increases in vitro.

Materials and Methods

Isolated rat, mouse and human colonic tissue mucosae and Caco-2 monolayers were exposed to cytochalasin D (cD) and sodium caprate (C10), in the absence and presence of the MLCK inhibitors, ML-9 and D PIK. Transepithelial electrical resistance (TEER) and Papp of [14C]-mannitol or FITC-dextran 4000 (FD-4) were measured. Western blots were used to measure MLC phosphorylation.

Results

Increases in Papp of [14C]-mannitol and decreases in TEER were induced by tight junction openers. These changes were attenuated by ML-9. D-PIK offset the FD-4 Papp increase induced by C10 in Caco-2 only, while ML-9 and PIK inhibited MLC directly. cD induced constriction of peri-junctional actin in Caco-2 monolayers, but this was prevented by ML-9. Although mannitol fluxes across colonic mucosae from dextran-sulphate (DSS)-treated mice were higher than control, they were not ameliorated by either ML-9 or PIK in vitro.

Conclusions

ML-9 inhibits paracellular permeability increases in several intestinal epithelial models. D-PIK reduced stimulated paracellular fluxes in Caco-2 monolayers, but not in tissue. Pre-established increases were not modified by two MLCK inhibitors in a mouse model of IBD.

Key words

caco-2 monolayers cytochalasins dextran sodium sulphate inflammatory bowel disease mouse models mannitol fluxes myosin light chain kinase transepithelial electrical resistance 

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Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Linda M. Feighery
    • 1
  • Sean W. Cochrane
    • 1
  • Teresa Quinn
    • 1
  • Alan W. Baird
    • 1
  • Daniel O’Toole
    • 1
  • Sian-Eleri Owens
    • 3
  • Diarmuid O’Donoghue
    • 2
  • Randall J. Mrsny
    • 3
  • David J. Brayden
    • 1
  1. 1.School of Agriculture, Food Science and Veterinary MedicineUniversity College DublinDublin 4Ireland
  2. 2.St. Vincent’s University Teaching HospitalDublin 4Ireland
  3. 3.Welsh School of PharmacyCardiff UniversityCardiffUK

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