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Phorbol Ester Treatment Increases Paracellular Permeability Across IEC-18 Gastrointestinal Epithelium In Vitro

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Abstract

The phorbol ester, TPA, transiently increases the transepithelial permeability across the gastrointestinal epithelium formed by IEC-18. There was a significant decrease in transepithelial resistance (RT) between 0 and 1.5 hr, accompanied by increased flux of polyethylene glycol (4000 MW), suggesting that the increase was across the tight junction. By 2 hr, the decrease in RT reversed and maintained control level. The transepithelial permeability increase was prevented by coincubation with the protein kinase C (PKC) inhibitor bisindolylmaleimide. There was a rapid (within 15 min) translocation of PKC-α from the cytosolic to the “membrane-associated” compartment, followed by a down-regulation that was detectable within 60 min of TPA treatment. The down-regulation of PKC-α from the membrane was prevented by either calpain inhibitor I or MG-132 and resulted in a sustained permeability increase. The permeability changes were not accompanied by significant effects on the amount or localization of the tight junctional proteins, occludin and ZO-1. However, occludin did show a reversible increase in phosphorylation with TPA treatment. Together these data support a role for PKC-α-mediated regulation of barrier permeability in an in vitro model of small intestinal epithelium, perhaps through modulation of the phosphorylation state of the tight junctional protein, occludin.

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  1. Lankenau Institute for Medical Research, Wynnewood, Pennsylvania

    C.W. Marano, L.A. Garulacan, N. Ginanni & J.M. Mullin

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  1. C.W. Marano
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  2. L.A. Garulacan
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  3. N. Ginanni
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  4. J.M. Mullin
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Marano, C., Garulacan, L., Ginanni, N. et al. Phorbol Ester Treatment Increases Paracellular Permeability Across IEC-18 Gastrointestinal Epithelium In Vitro. Dig Dis Sci 46, 1490–1499 (2001). https://doi.org/10.1023/A:1010696005958

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