Exogenous S-nitrosoglutathione attenuates inflammatory response and intestinal epithelial barrier injury in endotoxemic rats.

Abstract

Gut barrier injury in sepsis is a major contributor to distant organ dysfunction and bad clinical outcomes. Enteric glia-derived S-nitrosoglutathione (GSNO) has been recognized as a novel modulator of gut barrier integrity. In this study, we tested the potential therapeutic effect and mechanism of exogenous GSNO on endotoxin-induced inflammatory response and intestinal barrier injury in a rat model of endotoxemia. Male Sprague-Dawley rats were randomly assigned to four groups as follows: control (saline only), GSNO, lipopolysaccharide (LPS), and LPS + GSNO. Femoral venous injection of LPS (10 mg/kg) or saline was followed by GSNO (1 mg/kg) or saline injection 15 minutes later. Distal ileum tissues and blood were harvested after 3 hours of LPS/saline injection. The intestinal barrier function was measured histologically and by intestinal permeability to fluorescein isothiocyanate dextran. The ultrastructural change of the epithelial tight junction was observed using transmission electron microscope, and the expression level of tight junction protein ZO-1 was analyzed using immunofluorescence and Western blot. Systemic and intestinal inflammation was measured by analyzing the tumor necrosis factor and interleukin 1β levels in plasma and distal ileum tissue, respectively. The levels of nuclear factor κB (NF-κB) and myosin light-chain kinase in the distal ileum were measured by Western blot. Compared with the endotoxemic rats, the addition of GSNO reduced the intestinal injury observed in histologic sections, decreased permeability to fluorescein isothiocyanate dextran, attenuated damage of the junction between epithelia, and protected against the LPS-induced expression decrease of ZO-1. Furthermore, addition of GSNO reduced plasma and intestinal tumor necrosis factor and interleukin 1β levels as well as inhibited the LPS-induced up-regulation of myosin light-chain kinase expression and NF-κB p65 level in the intestine. The data indicate that GSNO protects against the LPS-induced systemic inflammatory response and attenuated intestinal inflammation and epithelial barrier injury in rats, possibly through the inhibition of the NF-κB pathway.