In addition, GSH also reduced the posttranslational modification of FOXA2. The levels of nitrosylated FOXA2 decreased sig nificantly by 40% and 76% www.selleckchem.com/products/CHIR-258.html in the presence of 0. 4 mM and 1 mM GSH, respectively. Collectively, these results suggest that higher levels of antioxidants in airway epithelial cells can reduce posttranslational modifi cation and inactivation of FOXA2 mediated by PCN generated ROS RNS. Furthermore, antioxidant treatment may enhance the expression of FOXA2. GSH treatment relieves the suppression of FOXA2 and represses mucin production induced by PCN Because the epithelial cells treated with GSH overcome the repression of FOXA2 expression and reduce post translational modification by PCN, we postulated that restored FOXA2 in turn, could inhibit the expression of MUC5AC and MUC5B mucins in the NCI H292 cells.

Western blot analyses showed that in the absence of GSH, PCN reduced the expression of FOXA2 by 50%, with corresponding 5 fold increase in MUC5AC and MUC5B expression. Addition of GSH significantly increased the expression of FOXA2, with cor responding decrease in the expression of MUC5AC and MUC5B mucins. Restored expression of FoxA2 is also associated with repression of MUC5AC and MUC5B transcription. Collectively, these results suggest that GSH effectively neutralizes PCN toxicity, restoring FOXA2 function, which in turn, may re press the transcription of MUC5AC and MUC5B genes as well as the expression of both mucins in airway epithelial cells. Discussion PCN is a redox active virulence factor of PA.

We have pre viously shown that PCN inhibits the expression of FOXA2 through the activation of pro GCHM signaling pathway Stat6 and EGFR. In this study, we demonstrate PCN generated ROS RNS causes posttranslational modifi cations nitrosylation, acetylation, and Batimastat ubiquitination of FOXA2, resulting in degradation of the transcriptional repressor of GCHM. Furthermore, FOXA2 modified by PCN generated ROS RNS has reduced binding capacity to the promoter of the MUC5B gene. The loss of FOXA2 ex pression is positively correlated to derepression of MUC5AC and MUC5B transcripts, as well as the overexpression of both mucins. Importantly, the antioxidant GSH neutralizes PCN mediated toxicity and reduces the nitrosylation and suppression of FOXA2 by PCN generated ROS RNS. Res toration of FOXA2 expression is positively correlated to the repression of both MUC5AC and MUC5B genes and mu cins.

Collectively, these results suggest that posttranslational modification and inactivation of FOXA2 induced by PCN generated ROS RNS may also contribute to GCHM and mucus hypersecretion. There has been a free copy continual debate as to the import ance of PCN to the pathogenesis of diseases in human airways, especially in CF. This is primarily because of conflicting levels of PCN that were recovered within a limited number of sputum samples from CF and non CF bronchiectatic patients.