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Am J Physiol Gastrointest Liver Physiol (February 21, 2008). doi:10.1152/ajpgi.00202.2007
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Submitted on May 5, 2007
Accepted on February 17, 2008

Probiotics Ameliorate the Hydrogen Peroxide-Induced Epithelial Barrier Disruption by a PKC and MAP Kinase-Dependent Mechanism

Ankur Seth1, Fang Yan2, D. Brent Polk3, and R.K. Rao4*

1 Physiology, University of Tennessee Health Science Center, Memphis, Tennessee, United States
2 Pediatrics, Vanderbilt University, Nashville, Tennessee, United States
3 Vanderbilt University; Pediatrics, Vanderbilt University, Nashville, Tennessee, United States
4 Physiology, University of Tennessee, Memphis, Tennessee, United States

* To whom correspondence should be addressed. E-mail: rkrao{at}physio1.utmem.edu.

Probiotics promote intestinal epithelial integrity and reduce infection and diarrhea. We evaluated the effect of Lactobacillus Rhamnosus GG-produced soluble proteins (P40 and P75) on the hydrogen peroxide-induced disruption of tight junctions and barrier function in Caco-2 cell monolayers. Pretreatment of cell monolayers with P40 or P75 attenuated the hydrogen peroxide-induced decrease in TER and increase in inulin permeability in a time and dose-dependent manner. P40 and P75 also prevented hydrogen peroxide-induced redistribution of occludin, ZO-1, E-cadherin and {beta}-catenin from the intercellular junctions and their dissociation from the detergent-insoluble fractions. Both P40 and P75 induced a rapid increase in the membrane translocation of PKC{beta}I and PKC{epsilon}. The attenuation of hydrogen peroxide-induced inulin permeability and redistribution of tight junction proteins by P40 and P75 was abrogated by Ro-32-0432, a PKC inhibitor. P40 and P75 also rapidly increased the levels of phospho-ERK1/2 in the detergent-insoluble fractions. U0126 (a MAP kinase inhibitor) attenuated the P40 and P75-mediated reduction of hydrogen peroxide-induced tight junction disruption and inulin permeability. These studies demonstrate that probiotic-secretory proteins protect the intestinal epithelial tight junctions and the barrier function from hydrogen peroxide-induced insult by a PKC and MAP kinase-dependent mechanism.




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