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Journal of Lipid Research, Vol. 47, 1416-1423, July 2006
Copyright © 2006 by American Society for Biochemistry and Molecular Biology

Arachidonic acid cascade and epithelial barrier function during Caco-2 cell differentiation

Raquel Martín-Venegas, Sònia Roig-Pérez, Ruth Ferrer and Juan José Moreno¹

Departament de Fisiologia, Facultat de Farmàcia, Universitat de Barcelona, 08028 Barcelona, Spain

Published, JLR Papers in Press, April 3, 2006.

¹ To whom correspondence should be addressed. e-mail: jjmoreno{at}ub.edu

The small intestinal epithelium is a highly dynamic system continuously renewed by a process involving cell proliferation and differentiation. The intestinal epithelium constitutes a permeability barrier regulating the vectorial transport of ions, water, and solutes. Morphological changes during cell differentiation, as well as changes in the activity of brush-border enzymes and the expression of transport proteins, are well established. However, little is known about the arachidonic acid (AA) cascade underlying epithelial cell differentiation or its role in the development of epithelial barrier function. The main purpose of this study was to examine the activity of the high-molecular-weight phospholipases A₂ (PLA₂) and cyclooxygenase (COX) pathway during differentiation, with particular emphasis on paracellular permeability. PLA₂ activity, AA release, COX-2 expression, prostaglandin E₂ (PGE₂) production, and paracellular permeability were studied in preconfluent, confluent, and differentiated Caco-2 cell cultures. Our results show that Caco-2 differentiation induces a decrease in both calcium-independent PLA₂ activity and COX-2 expression and, consequently, a decrease in AA release and PGE₂ synthesis in parallel with a reduction in paracellular permeability. Moreover, the addition of PGE₂ to differentiated cells, at concentrations similar to those detected in nondifferentiated cultures, induces the disruption of epithelial barrier function. These results suggest that AA release by calcium-independent PLA₂, COX-2 expression, and subsequent PGE₂ release are important for the maintenance of paracellular permeability in differentiated Caco-2 cells.

Abbreviations: AA, arachidonic acid; BEL, bromoenol lactone; COX, cyclooxygenase; cPLA₂, cytosolic phospholipase A₂; iPLA₂, Ca²⁺-independent phospholipase A₂; PGE₂, prostaglandin E₂; PLA₂, phospholipase A₂; sPLA₂, secretory phospholipase A₂; TEER, transepithelial electrical resistance

Supplementary key words intestine • tight junctions • phospholipase A₂ • cyclooxygenase • prostaglandins

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