AJP - Gastrointestinal and Liver Physiology, Vol 272, Issue 5 923-G934, Copyright © 1997 by American Physiological Society

ARTICLES

Hyperpermeability of intestinal epithelial monolayers is induced by NO: effect of low extracellular pH

N. Unno, M. J. Menconi, M. Smith, D. E. Aguirre and M. P. Fink
Department of Surgery, Beth Israel Deaconess Medical Center, Boston, Massachusetts, USA.

Nitric oxide (NO.) increases the permeability of Caco-2BBe enterocytic monolayers. Many of the toxic effects of NO. are thought to be mediated by the peroxynitrite anion (ONOO-), which, under mildly acidic conditions, can rearrange to yield an intermediate with reactivity similar to toxic OH.. Accordingly, we assessed the permeability of Caco-2BBe cells grown on permeable supports for 24 h in media titrated to normal or acidic extracellular pH (pHo) with or without the NO. donors 3-morpholinosydnonimine (SIN-1) or sodium nitroprusside (SNP). Incubation with 2 mM SIN-1 at pHo 6.8 or 0.6 mM SNP at pHo 6.5 increased permeability (apical-to-basolateral flux of fluorescein sulfonic acid), whereas at pHo 7.4 permeability was unaffected by these concentrations of NO. donors. Accumulation of NO2/NO3 in medium (index of NO. release) was not increased by incubation of cells with SIN-1 or SNP under mildly acidic conditions. Under acidic but not control conditions, incubation with SIN-1 caused disruption of perijunctional actin filaments as assessed by fluorescence microscopy. At pHo 6.8 and 6.5 (but not 7.4), SIN-1 significantly decreased intracellular levels of both ATP and glutathione. Incubation with 5 mM deferoxamine or 500 uM ascorbic acid (ONOO- scavengers) abrogated SIN-1-induced hyperpermeability. We conclude that mild acidosis augments NO.-induced intestinal epithelial permeability, possibly by promoting oxidant-mediated cytoskeletal damage and/or ATP depletion.

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