Differential effect of ethanol and hydrogen peroxide on barrier function and prostaglandin E2 release in differentiated Caco-2 cells: selective prevention by growth factors.
The present study investigates the effects of ethanol and hydrogen peroxide (H(2)O(2)) on the barrier function and prostaglandin E(2) (PGE(2)) release in differentiated Caco-2 cells. Epithelial barrier integrity was estimated by measuring transepithelial electrical resistance (TEER), the transport of reference compounds and lactate dehydrogenase leakage, the PGE(2) release by enzyme immunoassay. Ethanol and H(2)O(2) decreased TEER and increased the transport of lucifer yellow without affecting that of propranolol and phenylalanine. Only the effects of ethanol were accompanied by PGE(2) production and were reversible without causing long-term cytotoxicity. The cyclooxygenase-2 inhibitor, NS-398, prevented the effect of ethanol on both PGE(2) release and TEER, while inhibition of both cyclooxygenase-2 and tyrosine kinase drastically compromised cell viability and TEER recovery. Hepatocyte growth factor, keratinocyte growth factor or insulin prevented the effect of ethanol on cell permeability, but not on PGE(2) release. Their combination prevented the effect of H(2)O(2). In conclusion, ethanol and H(2)O(2) increased paracellular permeability in differentiated Caco-2 cells without affecting transcellular and active transport. Cyclooxygenase-2 stimulated PGE(2) release mediated the reversible effect of ethanol on tight junctions and, meanwhile, contributed to cell survival. Growth factors, normally present in the intestine, exerted a selective protective effect toward paracellular permeability increase induced by irritants.
Catalioto
Rose-Marie
RM
Pharmacology Department, Menarini Ricerche SpA, Florence, Italy. chimfarm@menarini-ricerche.it
Festa
Carla
C
Triolo
Antonio
A
Altamura
Maria
M
Maggi
Carlo Alberto
CA
Giuliani
Sandro
S
D018938:Caco-2 Cells; D002454:Cell Differentiation; D002470:Cell Survival; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D015232:Dinoprostone / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D017097:Electric Impedance; D004847:Epithelial Cells / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D000431:Ethanol / Q000633:toxicity*; D051523:Fibroblast Growth Factor 7 / Q000378:metabolism; D005346:Fibroblast Growth Factors / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D017228:Hepatocyte Growth Factor / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity*; D007328:Insulin / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism*; D007509:Irritants / Q000633:toxicity*; D007546:Isoquinolines / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D010539:Permeability; D010649:Phenylalanine / Q000378:metabolism; D011433:Propranolol / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology
D016861:Cyclooxygenase Inhibitors; D005456:Fluorescent Dyes; D007328:Insulin; D036341:Intercellular Signaling Peptides and Proteins; D007509:Irritants; D007546:Isoquinolines; D047428:Protein Kinase Inhibitors; D051523:Fibroblast Growth Factor 7; D000431:Ethanol; D010649:Phenylalanine; D005346:Fibroblast Growth Factors; D017228:Hepatocyte Growth Factor; C017475:lucifer yellow; D011433:Propranolol; D006861:Hydrogen Peroxide; D007770:L-Lactate Dehydrogenase; D015232:Dinoprostone