title: Protective action of NADPH oxidase inhibitors and role of NADPH oxidase in pathogenesis of colon inflammation in mice.
abstract: OBJECTIVE
  To investigate the role of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in colon epithelial cells in the pathogenesis of acute and chronic colon inflammation in a mouse model of dextran sulphate sodium (DSS)-induced colitis.
  
  
  METHODS
  Balb/c mice were divided into three groups: 8 mice with acute DSS-induced colitis (3.5% DSS solution; 7 d), 8 mice with chronic DSS-induced colitis (3.5% DSS solution for 5 d + water for 6 d; 4 cycles; total: 44 d) and 12 mice without DSS supplementation as a control group. Primary colonic epithelial cells were isolated using chelation method. The cells were cultivated in the presence of mediators (lipopolysaccharide (LPS), apocynin or diphenyleneiodonium). Viability of cells was assessed by fluorescent microscopy. Production of reactive oxygen species (ROS) by the cells was measured fluorometrically using Amplex Red. Production of tumour necrosis factor-alpha (TNF-α) by the colonic epithelial cells was analysed by ELISA. Nox1 gene expression was assessed by real-time PCR.
  
  
  RESULTS
  Our study showed that TNF-α level was increased in unstimulated primary colonic cells both in the acute and chronic colitis groups, whereas decreased viability, increased ROS production, and expression of Nox1 was characteristic only for chronic DSS colitis mice when compared to the controls. The stimulation by LPS increased ROS generation via NADPH oxidase and decreased cell viability in mice with acute colitis. Treatment with NADPH oxidase inhibitors increased cell viability and decreased the levels of ROS and TNF-α in the LPS-treated cells isolated from mice of both acute and chronic colitis groups.
  
  
  CONCLUSIONS
  Our study revealed the importance of NADPH oxidase in the pathogenesis of both acute and chronic inflammation of the colon.
authors.affiliation: Rima Ramonaite, Jurgita Skieceviciene, Simonas Juzenas, Violeta Salteniene, Juozas Kupcinskas, Limas Kupcinskas, Institute for Digestive Research, Academy of Medicine, Lithuanian University of Health Sciences, Kaunas LT 44307, Lithuania.
authors.forename: Rima
authors.initials: R
authors.lastname: Ramonaite
authors.affiliation: Rima Ramonaite, Jurgita Skieceviciene, Simonas Juzenas, Violeta Salteniene, Juozas Kupcinskas, Limas Kupcinskas, Institute for Digestive Research, Academy of Medicine, Lithuanian University of Health Sciences, Kaunas LT 44307, Lithuania.
authors.forename: Jurgita
authors.initials: J
authors.lastname: Skieceviciene
authors.affiliation: Rima Ramonaite, Jurgita Skieceviciene, Simonas Juzenas, Violeta Salteniene, Juozas Kupcinskas, Limas Kupcinskas, Institute for Digestive Research, Academy of Medicine, Lithuanian University of Health Sciences, Kaunas LT 44307, Lithuania.
authors.forename: Simonas
authors.initials: S
authors.lastname: Juzenas
authors.affiliation: Rima Ramonaite, Jurgita Skieceviciene, Simonas Juzenas, Violeta Salteniene, Juozas Kupcinskas, Limas Kupcinskas, Institute for Digestive Research, Academy of Medicine, Lithuanian University of Health Sciences, Kaunas LT 44307, Lithuania.
authors.forename: Violeta
authors.initials: V
authors.lastname: Salteniene
authors.affiliation: Rima Ramonaite, Jurgita Skieceviciene, Simonas Juzenas, Violeta Salteniene, Juozas Kupcinskas, Limas Kupcinskas, Institute for Digestive Research, Academy of Medicine, Lithuanian University of Health Sciences, Kaunas LT 44307, Lithuania.
authors.forename: Juozas
authors.initials: J
authors.lastname: Kupcinskas
authors.affiliation: Rima Ramonaite, Jurgita Skieceviciene, Simonas Juzenas, Violeta Salteniene, Juozas Kupcinskas, Limas Kupcinskas, Institute for Digestive Research, Academy of Medicine, Lithuanian University of Health Sciences, Kaunas LT 44307, Lithuania.
authors.forename: Paulius
authors.initials: P
authors.lastname: Matusevicius
authors.affiliation: Rima Ramonaite, Jurgita Skieceviciene, Simonas Juzenas, Violeta Salteniene, Juozas Kupcinskas, Limas Kupcinskas, Institute for Digestive Research, Academy of Medicine, Lithuanian University of Health Sciences, Kaunas LT 44307, Lithuania.
authors.forename: Vilmante
authors.initials: V
authors.lastname: Borutaite
authors.affiliation: Rima Ramonaite, Jurgita Skieceviciene, Simonas Juzenas, Violeta Salteniene, Juozas Kupcinskas, Limas Kupcinskas, Institute for Digestive Research, Academy of Medicine, Lithuanian University of Health Sciences, Kaunas LT 44307, Lithuania.
authors.forename: Limas
authors.initials: L
authors.lastname: Kupcinskas
mesh_terms: D000098:Acetophenones / Q000494:pharmacology*; D000208:Acute Disease; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D002478:Cells, Cultured; D002908:Chronic Disease; D003092:Colitis / Q000139:chemically induced / Q000201:enzymology / Q000473:pathology / Q000517:prevention & control*; D003106:Colon / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D016264:Dextran Sulfate; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004847:Epithelial Cells / Q000187:drug effects / Q000201:enzymology / Q000473:pathology; D006861:Hydrogen Peroxide / Q000378:metabolism; D018836:Inflammation Mediators / Q000378:metabolism; D007413:Intestinal Mucosa / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D008297:Male; D008807:Mice, Inbred BALB C; D009247:NADH, NADPH Oxidoreductases / Q000037:antagonists & inhibitors / Q000378:metabolism; D000074624:NADPH Oxidase 1; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D000098:Acetophenones; D000893:Anti-Inflammatory Agents; D004791:Enzyme Inhibitors; D018836:Inflammation Mediators; D014409:Tumor Necrosis Factor-alpha; D016264:Dextran Sulfate; C056165:acetovanillone; D006861:Hydrogen Peroxide; D009247:NADH, NADPH Oxidoreductases; D000074624:NADPH Oxidase 1; D019255:NADPH Oxidases; C000614711:NOX1 protein, mouse
keywords: Apocynin; Dextran sulphate sodium-induced colitis; Diphenyleneiodonium; Inflammatory bowel disease, Nicotinamide adenine dinucleotide phosphate oxidase; Reactive oxygen species; Tumour necrosis factor-α