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High glucose concentration in isotonic media alters Caco-2 cell permeability

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Abstract

Caco-2 cell permeability was evaluated in isotonic media containing high (25mM) or physiological (5.5mM) glucose concentrations. Transepithelial electrical resistance (TEER) and membrane fluidity were measured to assess glucose-induced alterations in physical barrier properties. In parallel, distribution of the actin filament (F-actin) and zonula occludens-1 (ZO-1) proteins was assessed by confocal microscopy. Transepithelial fluxes of mannitol, hydrocortisone, digoxin, and glycyl sarcosine (Gly-Sar) that permeate the intestinal mucosa by various pathways were measured to quantify the effect of glucose-induced changes on Caco-2 cell permeability. High glucose decreased maximum TEER of cell monolayers by 47%, whereas membrane fluidity at the hydrophobic core and lipid/polar head interphase was significantly increased. F-actin distribution in high glucose cells appeared more diffuse while ZO-1 was unchanged. Mannitol and hydrocortisone fluxes across Caco-2 cells cultured in high glucose increased by 65% and 24%, respectively. In addition, high glucose decreased the maximum transport capacity (Vmax) of PepT-1. P-glycoprotein activity, however, was unchanged. In conclusion, high extracellular glucose concentration in isotonic media significantly alters physical barrier properties of Caco-2 cell monolayers, which predominantly affects transepithelial transport of solutes permeating the cell barrier by paracellular and transcellular passive diffusion and facilitated transport mediated by the proton-dependent oligopeptide transporter (PepT-1).

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Authors and Affiliations

  1. Division of Pharmaceutical Sciences, College of Pharmacy, University of Cincinnati, Cincinnati, OH

    Vanessa M. D Souza & Giovanni M. Pauletti

  2. Department of Environmental Health and Center for Environmental Genetics, University of Cincinnati Medical Center, Cincinnati, OH

    Howard G. Shertzer

  3. Department of Molecular Genetics, Biochemistry, and Microbiology, University of Cincinnati, College of Medicine, Cincinnati, OH

    Anil G. Menon

Authors
  1. Vanessa M. D Souza
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  2. Howard G. Shertzer
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  3. Anil G. Menon
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  4. Giovanni M. Pauletti
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Correspondence to Giovanni M. Pauletti.

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Published: August 14, 2003

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Souza, V.M.D., Shertzer, H.G., Menon, A.G. et al. High glucose concentration in isotonic media alters Caco-2 cell permeability. AAPS PharmSci 5, 24 (2003). https://doi.org/10.1208/ps050324

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  • DOI: https://doi.org/10.1208/ps050324

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