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J. Biol. Chem., Vol. 283, Issue 6, 3574-3583, February 8, 2008
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Role of Phospholipase C
-induced Activation of Protein Kinase C
(PKC) and PKCβI in Epidermal Growth Factor-mediated Protection of Tight Junctions from Acetaldehyde in Caco-2 Cell Monolayers*
From the Department of Physiology, University of Tennessee Health Science Center, Memphis, Tennessee 38163
Epidermal growth factor (EGF) protects the intestinal epithelial tight junctions from acetaldehyde-induced insult. The role of phospholipase C
(PLC) and protein kinase C (PKC) isoforms in the mechanism of EGF-mediated protection of tight junction from acetaldehyde was evaluated in Caco-2 cell monolayers. EGF-mediated prevention of acetaldehyde-induced decrease in transepithelial electrical resistance and an increase in inulin permeability, and subcellular redistribution of occludin and ZO-1 was attenuated by reduced expression of PLC1 by short hairpin RNA. EGF induced a rapid activation of PLC1 and PLC-dependent membrane translocation of PKC
and PKCβI. Inhibition of PKC activity or selective interference of membrane translocation of PKC and PKCβI by RACK interference peptides attenuated EGF-mediated prevention of acetaldehyde-induced increase in inulin permeability and redistribution of occludin and ZO-1. BAPTA-AM and thapsigargin blocked EGF-induced membrane translocation of PKCβI and attenuated EGF-mediated prevention of acetaldehyde-induced disruption of tight junctions. EGF-induced translocation of PKC and PKCβI was associated with organization of F-actin near the perijunctional region. This study shows that PLC-mediated activation of PKC and PKCβI and intracellular calcium is involved in EGF-mediated protection of tight junctions from acetaldehyde-induced insult.
Received for publication, November 7, 2007
* This work was supported by National Institutes of Health Grants R01-DK55532 and R01-AA12307. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
1 To whom correspondence should be addressed: Dept. of Physiology, University of Tennessee, 894 Union Ave., Memphis, TN 38163. Tel.: 901-448-3235; Fax: 901-448-7126; E-mail: rkrao{at}physio1.utmem.edu.
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