NF-kappa B activation by oxidative stress and inflammation suppresses contractility in colonic circular smooth muscle cells.

BACKGROUND & AIMS: Transcription factor nuclear factor kappa B (NF-kappa B) plays a critical role in transcriptional changes in several diseases, including inflammation. The aim of this study was to investigate whether NF-kappa B is activated by inflammation and oxidative stress in colonic circular smooth muscle cells and whether that leads to suppression of their contractility.
METHODS: The experiments were performed on freshly dissociated single cells using electrophoretic mobility shift assay, Western immunoblotting, and immunofluorescence imaging.
RESULTS: The NF-kappa B DNA binding was approximately 6-fold greater in cells from the inflamed colon vs. those from the normal colon. Supershift assay indicated that the antibodies to p65, p50, and c-Rel, but not that to p52, shifted the NF-kappa B band. Western immunoblotting and immunofluorescence imaging also demonstrated the presence of p65, p50, and c-Rel proteins in the cytoplasm and their translocation to the nucleus by H(2)O(2)-induced oxidative stress. H(2)O(2) treatment degraded I kappa B(beta), but not I kappa B(alpha), to translocate NF-kappa B to the nucleus. Hydrogen peroxide concentration and time dependently activated NF-kappa B DNA binding and suppressed cell contraction to acetylcholine. NF-kappa B inhibitors significantly inhibited these effects. Inhibition of NF-kappa B prior to and during inflammation in intact dogs also reversed the suppression of contractility.
CONCLUSIONS: Transcription factor NF-kappa B is activated in colonic circular muscle cells by inflammation and oxidative stress. This activation of NF-kappa B mediates the suppression of cell contractility.