Acute lung injury (ALI) is a state of inflammation that breaks down the lung endothelial and epithelial cell barriers. In the current study, we investigated the role of resveratrol (RES) in regulating the expression and functions of tight junction proteins (TJP) in epithelial cell responses following exposure to this superantigen. To this end, C3H mice were given resveratrol orally twice prior to intranasal challenge with lethal SEB doses. 16S rRNA results showed that there were microbes transported in the blood in addition to the lung and colonic tissues. For this purpose, we used a reporter E. coli-GFP labeled bacterium to monitor and examine the viability of this bacterium in case of leak from lung into blood and other body compartments during the peak of ALI. The flow cytometry results showed significant reduction in copy numbers of this reporter in the blood in RES-treated mice in comparison to vehicle-treated mice. However, bacteriological examination for viability by culture of the blood showed all were negative for all experimental groups. Mouse transcriptome array showed increase in TJP gene expression in colonic and alveolar epithelial cells of RES-treated mice. Some of these genes were validated with qPCR. We found that there was an increase in the expression of some TJP genes following RES treatment. Commensals and probiotics are also known to decrease intestinal barrier dysfunction caused by inflammatory cytokines. Therefore, mass spectrometry (MS) analysis showed increase in the metabolome in colons of RES-treated group that help in strengthening of intestinal barrier. In conclusion, RES treatment increased directly or indirectly the rigidity of cell boundaries resulting in amelioration of ALI-dependent complications.