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Articles in PresS, published online ahead of print September 25, 2002
Am J Physiol Gastrointest Liver Physiol, 10.1152/ajpgi.00121.2002
Submitted on March 27, 2002
Accepted on September 10, 2002
1 Department of Clinical Sciences, North Carolina State University, Raleigh, NC, USA
2 Department of Molecular and Biological Sciences, North Carolina State University, Raleigh, NC, USA
* To whom correspondence should be addressed. E-mail: Anthony_Blikslager{at}ncsu.edu.
We have previously shown that PGE2 and PGI2 induce recovery of transepithelial resistance (TER) in ischemia-injured porcine ileal mucosa, associated with initial increases in Cl- secretion. We believe that the latter generates an osmotic gradient that stimulates re-sealing of tight junctions. Because of evidence implicating PI3'K in regulating tight junction assembly, we postulated that this signaling pathway is involved in PG-induced mucosal recovery. Porcine ileum was subjected to 45-minutes ischemia, after which TER was monitored for a 180-minute recovery period. Endogenous prostaglandin production was inhibited with indomethacin (5µM). PGE2 (1µM) and PGI2 (1µM) stimulated recovery of TER, which was inhibited by serosal application of the osmotic agent urea (300mOsm). The PI3'K inhibitor wortmannin (10nM) blocked recovery of TER in response to PGs or mucosal urea. Immunofluorescence imaging of recovering epithelium revealed that PGs restored occludin and ZO-1 distribution to inter-epithelial junctions, and this pattern was disrupted by pre-treatment with wortmannin. These experiments suggest PGs stimulate recovery of paracellular resistance via a mechanism involving transepithelial osmotic gradients and PI3'K-dependent restoration of tight junction protein distribution.
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