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Zinc Supplementation Modifies Tight Junctions and Alters Barrier Function of CACO-2 Human Intestinal Epithelial Layers

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Abstract

Background

Zinc deficiency is known to result in epithelial barrier leak in the GI tract. Precise effects of zinc on epithelial tight junctions (TJs) are only beginning to be described and understood. Along with nutritional regimens like methionine-restriction and compounds such as berberine, quercetin, indole, glutamine and rapamycin, zinc has the potential to function as a TJ modifier and selective enhancer of epithelial barrier function.

Aims

The purpose of this study was to determine the effects of zinc-supplementation on the TJs of a well-studied in vitro GI model, CACO-2 cells.

Methods

Barrier function was assessed electrophysiologically by measuring transepithelial electrical resistance (Rt), and radiochemically, by measuring transepithelial (paracellular) diffusion of 14C-D-mannitol and 14C-polyethyleneglycol. TJ composition was studied by Western immunoblot analyses of occludin, tricellulin and claudins-1 to -5 and -7.

Results

Fifty- and 100-μM zinc concentrations (control medium is 2 μM) significantly increase Rt but simultaneously increase paracellular leak to D-mannitol. Claudins 2 and 7 are downregulated in total cell lysates, while occludin, tricellulin and claudins-1, -3, -4 and -5 are unchanged. Claudins-2 and -7 as well as tricellulin exhibit decreased cytosolic content as a result of zinc supplementation.

Conclusions

Zinc alters CACO-2 TJ composition and modifies TJ barrier function selectively. Zinc is one of a growing number of “nutraceutical” substances capable of enhancing epithelial barrier function, and may find use in countering TJ leakiness induced in various disease states.

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Abbreviations

TJ:

Tight junction

PD:

Potential difference

Iscc:

Short circuit current

Rt :

Transepithelial electrical resistance

MT 1/2:

Metallothionein 1/2

GI:

Gastrointestinal

FBS:

Fetal bovine serum

PBS:

Phosphate buffered saline

RDA:

Recommended daily allowance

SEM:

Standard error of the mean

LDH:

Lactate dehydrogenase

XTT:

Tetrazolium dye, sodium 2,3,-bis(2-methoxy-4-nitro-5-sulfophenyl-5-[(phenylamino)-carbonyl]-2H-tetrazolium)

LSC:

Liquid scintillation counting

PEG:

Polyethyleneglycol

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Acknowledgments

This research was funded in part by a grant award from the Sharpe-Strumia Foundation. We are very appreciative of the editorial assistance of Ms. Kristin M. Hayden in preparation of this manuscript.

Conflict of interest

None.

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Authors and Affiliations

  1. Lankenau Institute for Medical Research, 100 W Lancaster Ave, Wynnewood, PA, 19096, USA

    Xuexuan Wang, Mary Carmen Valenzano, Joanna M. Mercado & James M. Mullin

  2. Department of Chemistry, Saint Joseph’s University, 5600 City Avenue, Philadelphia, PA, 19131, USA

    E. Peter Zurbach

  3. Division of Gastroenterology, Lankenau Medical Center, Wynnewood, PA, 19096, USA

    James M. Mullin

Authors
  1. Xuexuan Wang
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  2. Mary Carmen Valenzano
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  3. Joanna M. Mercado
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  5. James M. Mullin
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Correspondence to James M. Mullin.

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Wang, X., Valenzano, M.C., Mercado, J.M. et al. Zinc Supplementation Modifies Tight Junctions and Alters Barrier Function of CACO-2 Human Intestinal Epithelial Layers. Dig Dis Sci 58, 77–87 (2013). https://doi.org/10.1007/s10620-012-2328-8

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