Nitric oxide dilates tight junctions and depletes ATP in cultured Caco-2BBe intestinal epithelial monolayers.
We tested the hypothesis that nitric oxide (NO) modulates the permeability of tight junctions in a model intestinal epithelium (Caco-2BBe monolayers). Incubation with sodium nitroprusside (SNP) resulted in time- and concentration-dependent decreases in transepithelial resistance. Permeability to fluorescein sulfonic acid increased during incubation for 24 h in the presence of 1.25 mM SNP, 5 mM S-nitroso-N-acetylpenicillamine (SNAP), or 1% NO gas. SNP-induced hyperpermeability was not due to loss of cell viability, as confirmed by intact ultrastructure, unaltered lactate dehydrogenase release, and ability to recover baseline permeability. Incubation with SNP increased permeability but only minimally increased intracellular levels of guanosine 3',5'-cyclic monophosphate (cGMP). Incubation with Escherichia coli heat-stable enterotoxin greatly increased cGMP levels with only a minimal effect on permeability. Cellular ATP levels decreased after incubation with SNP, SNAP, or gaseous NO. Incubation with SNP led to diminished fluoresceinphalloidin staining of junctional actin (confocal microscopy) and widened tight junctions (electron microscopy). We conclude that NO reduces ATP levels and reversibly increases the permeability of tight junctions in cultured Caco-2BBe cells.
Salzman
A L
AL
Department of Surgery, Beth Israel Hospital, Harvard Medical School, Boston, Massachusetts 02215.
Menconi
M J
MJ
Unno
N
N
Ezzell
R M
RM
Casey
D M
DM
Gonzalez
P K
PK
Fink
M P
MP
D000255:Adenosine Triphosphate / Q000037:antagonists & inhibitors*; D006152:Cyclic GMP / Q000378:metabolism; D003847:Deoxyglucose / Q000494:pharmacology; D004553:Electric Conductivity; D006801:Humans; D007365:Intercellular Junctions / Q000187:drug effects / Q000502:physiology*; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D007425:Intracellular Membranes / Q000378:metabolism; D046911:Macromolecular Substances; D009569:Nitric Oxide / Q000494:pharmacology / Q000502:physiology*; D009599:Nitroprusside / Q000494:pharmacology; D010396:Penicillamine / Q000031:analogs & derivatives / Q000494:pharmacology; D010539:Permeability / Q000187:drug effects; D026423:S-Nitroso-N-Acetylpenicillamine; D014407:Tumor Cells, Cultured; D014665:Vasodilator Agents / Q000494:pharmacology
D046911:Macromolecular Substances; D014665:Vasodilator Agents; D009599:Nitroprusside; D009569:Nitric Oxide; D026423:S-Nitroso-N-Acetylpenicillamine; D000255:Adenosine Triphosphate; D003847:Deoxyglucose; D010396:Penicillamine; D006152:Cyclic GMP