The role of the glutathione antioxidant system in gut barrier failure in a rodent model of experimental necrotizing enterocolitis

Surgery
Natasha KellyJeffrey S Upperman

Abstract

Evidence suggests that intestinal barrier failure in necrotizing enterocolitis results in part from overproduction of nitric oxide and other toxic oxidant species that result in enterocyte death and intestinal barrier failure. We hypothesize that the glutathione detoxifying system is important in maintaining intestinal barrier integrity by protecting against nitrosative stress. Newborn rats were subjected to hypoxia (5% O2, tid) and fed formula by gavage (NEC), or were breast-fed without hypoxia (BF). Rats were killed and the distal ilea were harvested for RNA, protein, and morphologic studies. RNA underwent cDNA microarray analysis. To assess glutathione in protecting against nitrosative stress, IEC-6 cells were exposed to SIN-1 and/or L-buthionine-(S,R)-sulfoximine (BSO), a glutathione inhibitor. Cells were analyzed for glutathione-S-transferase activity, apoptosis and mitochondrial function. BF controls developed normal intestinal architecture, whereas NEC animals sustained damage to the intestinal epithelium. Microarray analysis demonstrated that 93 genes were overexpressed in NEC compared with controls. In the array, glutathione-S-transferase pi and alpha 2, GSH-dependent detoxifying enzymes, RNA were upregulated compared ...Continue Reading

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