title: Oxidative stress disrupts purinergic neuromuscular transmission in the inflamed colon.
abstract: Colitis, induced by trinitrobenzene sulfonic acid (TNBS) in guinea pig, leads to decreased purinergic neuromuscular transmission resulting in a reduction in inhibitory junction potentials (IJPs) in colonic circular muscle. We explored possible mechanisms responsible for this inflammation-induced neurotransmitter plasticity. Previous studies have suggested that the deficit in inflamed tissue involves decreased ATP release. We therefore hypothesized that decreased purinergic transmission results from inflammation-induced free radical damage to mitochondria, leading to decreased purine synthesis and release. Stimulus-induced release of purines was measured using high-performance liquid chromatography, and quantities of all purines measured were significantly reduced in the inflamed colons as compared to controls. To test whether decreased mitochondrial function affects the IJP, colonic muscularis preparations were treated with the mitochondrial ATP synthase inhibitors oligomycin or dicyclohexylcarbodiimide, which resulted in a significant reduction of IJP amplitude. Induction of oxidative stress in vitro, by addition of H2O2 to the preparation, also significantly reduced IJP amplitude. Purinergic neuromuscular transmission was significantly restored in TNBS-inflamed guinea pigs, and in dextran sodium sulfate-inflamed mice, treated with a free radical scavenger. Furthermore, propulsive motility in the distal colons of guinea pigs with TNBS colitis was improved by in vivo treatment with the free radical scavenger. We conclude that oxidative stress contributes to the reduction in purinergic neuromuscular transmission measured in animal models of colitis, and that these changes can be prevented by treatment with a free radical scavenger, resulting in improved motility.
authors.affiliation: Department of Neurological Sciences, University of Vermont, Burlington, VT 05405, USA.
authors.forename: Jane A
authors.initials: JA
authors.lastname: Roberts
authors.forename: Leonie
authors.initials: L
authors.lastname: Durnin
authors.forename: Keith A
authors.initials: KA
authors.lastname: Sharkey
authors.forename: Violeta N
authors.initials: VN
authors.lastname: Mutafova-Yambolieva
authors.forename: Gary M
authors.initials: GM
authors.lastname: Mawe
mesh_terms: D000818:Animals; D003092:Colitis / Q000139:chemically induced / Q000378:metabolism / Q000503:physiopathology*; D003497:Cyclic N-Oxides / Q000494:pharmacology; D016264:Dextran Sulfate; D005260:Female; D016166:Free Radical Scavengers / Q000494:pharmacology; D005769:Gastrointestinal Motility / Q000187:drug effects; D006168:Guinea Pigs; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009130:Muscle, Smooth / Q000502:physiology*; D018384:Oxidative Stress; D011687:Purines / Q000378:metabolism*; D013113:Spin Labels; D009435:Synaptic Transmission; D014302:Trinitrobenzenesulfonic Acid
chemical_list: D003497:Cyclic N-Oxides; D016166:Free Radical Scavengers; D011687:Purines; D013113:Spin Labels; D014302:Trinitrobenzenesulfonic Acid; D016264:Dextran Sulfate; C001803:tempol