Cystic fibrosis gene mutation reduces epithelial cell acidification and injury in acid-perfused mouse duodenum.
OBJECTIVE
Dysfunction of the cystic fibrosis transmembrane regulator (CFTR) is associated with diminished duodenal HCO3- secretion, despite a reported lack of clinical duodenal ulceration in affected subjects. We hypothesized that duodenal epithelial cells expressing a mutant CFTR have enhanced resistance to acid-induced injury. To test this hypothesis, we measured duodenal epithelial cell intracellular pH (pHi), injury, and acid back-diffusion in response to a luminal acid challenge in transgenic mice.


METHODS
A murine colony was established for the CFTR DeltaF508 (DeltaF) mutation. Epithelial cell pH i was measured by microscopy with a trapped, fluorescent pH-sensitive dye in living C57BL/6 and DeltaF/DeltaF, +/DeltaF, and +/+ mice. In vivo confocal microscopy confirmed the localization of the dye in the cytoplasm of the epithelial cells. Epithelial injury was measured fluorometrically using propidium iodide. Duodenal epithelial bicarbonate secretion and proton permeability were measured by back-titration. Bicarbonate secretion and acid back-diffusion were measured in a perfused duodenal loop.


RESULTS
Basal and post-acid exposure bicarbonate secretion were reduced in DeltaF/DeltaF mice, although acid back-diffusion was similar to controls. Epithelial pHi of CFTR DeltaF/DeltaF mice during luminal acid exposure was significantly higher than pHi in +/DeltaF, +/+, or C57BL/6 mice. Acid-related epithelial injury was markedly less in DeltaF/DeltaF mice in comparison with the other groups.


CONCLUSIONS
Increased cellular buffering power of the epithelial cells of DeltaF/DeltaF mice likely protects against acidification and injury during acid exposure. We speculate that this protective mechanism partially underlies the perceived relative lack of peptic ulceration in patients affected by cystic fibrosis.
Hirokawa
Masahiko
M
Greater Los Angeles Veterans Affairs Healthcare System, California 90073, USA.
Takeuchi
Tetsu
T
Chu
Sahaoyou
S
Akiba
Yasutada
Y
Wu
Vincent
V
Guth
Paul H
PH
Engel
Eli
E
Montrose
Marshall H
MH
Kaunitz
Jonathan D
JD
D000818:Animals; D001639:Bicarbonates / Q000378:metabolism; D002712:Chlorides / Q000378:metabolism; D003550:Cystic Fibrosis / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D004058:Diffusion; D004386:Duodenum / Q000378:metabolism* / Q000473:pathology; D006863:Hydrogen-Ion Concentration; D007413:Intestinal Mucosa / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D009154:Mutation; D000074083:Sodium-Hydrogen Exchanger 3; D017923:Sodium-Hydrogen Exchangers / Q000037:antagonists & inhibitors
D001639:Bicarbonates; D002712:Chlorides; D000074083:Sodium-Hydrogen Exchanger 3; D017923:Sodium-Hydrogen Exchangers; D019005:Cystic Fibrosis Transmembrane Conductance Regulator