title: Chemiluminescence assay of mucosal reactive oxygen metabolites in inflammatory bowel disease.
abstract: Previous studies suggesting increased reactive oxygen metabolite (ROM) production in inflammatory bowel disease have been restricted to peripheral blood and isolated intestinal phagocytes. In the current study, chemiluminescence and the effect of various scavengers, enzymes, and enzyme inhibitors were used to show that ROMs account for the increased production of oxidants by colorectal mucosal biopsy specimens in inflammatory bowel disease. Luminol-amplified chemiluminescence was increased in active ulcerative colitis [macroscopic grade 1: 25 photons.mg-1.min.10(-3) (median), 8-47 (95% confidence intervals), n = 40; grade 2: 89, 65-156, n = 30; grade 3: 247, 133-562, n = 13] and Crohn's disease [mild: 9, 3-84, n = 6; severe: 105, 25-789 (range), n = 5] compared with normal-looking mucosa (ulcerative colitis: 0.8, 0.4-1.4, n = 22, P less than 0.01; Crohn's disease: 0.8, 0.1-2, n = 6, P less than 0.05) and controls (0.6, 0.04-1.4, n = 52, P less than 0.01). In ulcerative colitis, luminol chemiluminescence correlated with microscopic inflammation (Spearman's p = 0.74, P = 0.0001) and was decreased by sodium azide (-89%, P less than 0.05), taurine (-31%, P less than 0.05), catalase (-23%, P less than 0.05), and dimethyl sulfoxide (-29%, P less than 0.05). Superoxide dismutase and oxypurinol decreased lucigenin chemiluminescence in ulcerative colitis by -63% (P less than 0.05) and -27% (P less than 0.05), respectively. Luminol chemiluminescence correlated with lucigenin chemiluminescence (Spearman's rho = 0.72, P = 0.003). These results suggest that neutrophil-derived oxidants (superoxide, hydrogen peroxide, hydroxyl radical, and hypochlorite) are generated in colorectal mucosa in active inflammatory bowel disease and support the hypothesis that production of such metabolites by neutrophils is of major pathogenetic importance.
authors.affiliation: Gastrointestinal Science Research Unit, London Hospital Medical College, England.
authors.forename: N J
authors.initials: NJ
authors.lastname: Simmonds
authors.forename: R E
authors.initials: RE
authors.lastname: Allen
authors.forename: T R
authors.initials: TR
authors.lastname: Stevens
authors.forename: R N
authors.initials: RN
authors.lastname: Van Someren
authors.forename: D R
authors.initials: DR
authors.lastname: Blake
authors.forename: D S
authors.initials: DS
authors.lastname: Rampton
mesh_terms: D000166:Acridines; D000328:Adult; D000368:Aged; D005260:Female; D016166:Free Radical Scavengers; D006801:Humans; D015212:Inflammatory Bowel Diseases / Q000378:metabolism* / Q000473:pathology; D007413:Intestinal Mucosa / Q000378:metabolism*; D008163:Luminescent Measurements; D008165:Luminol; D008297:Male; D008875:Middle Aged; D016877:Oxidants / Q000378:metabolism*; D009195:Peroxidase / Q000037:antagonists & inhibitors / Q000378:metabolism; D012907:Smoking
chemical_list: D000166:Acridines; D016166:Free Radical Scavengers; D016877:Oxidants; C033472:10,10'-dimethyl-9,9'-biacridinium; D008165:Luminol; D009195:Peroxidase