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

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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.

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Acknowledgments

This work was funded by Irish Health Research Board Grant RP/86/2002 and Science Foundation Investigator Grant 04/IN3/B575 to David Brayden. Thanks also to Professor Paul Moynagh at National University of Ireland, Maynooth, for assistance with the MLC Western blots.

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Authors and Affiliations

  1. School of Agriculture, Food Science and Veterinary Medicine, University College Dublin, Belfield, Dublin 4, Ireland

    Linda M. Feighery, Sean W. Cochrane, Teresa Quinn, Alan W. Baird, Daniel O’Toole & David J. Brayden

  2. St. Vincent’s University Teaching Hospital, Elm Park, Dublin 4, Ireland

    Diarmuid O’Donoghue

  3. Welsh School of Pharmacy, Cardiff University, King Edward VII Avenue, Cardiff, Wales, UK

    Sian-Eleri Owens & Randall J. Mrsny

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  1. Linda M. Feighery
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Correspondence to David J. Brayden.

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Feighery, L.M., Cochrane, S.W., Quinn, T. et al. Myosin Light Chain Kinase Inhibition: Correction of Increased Intestinal Epithelial Permeability In Vitro . Pharm Res 25, 1377–1386 (2008). https://doi.org/10.1007/s11095-007-9527-6

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  • DOI: https://doi.org/10.1007/s11095-007-9527-6

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