Abstract | AIM: 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. CONCLUSION:
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Authors | Rima Ramonaite, Jurgita Skieceviciene, Simonas Juzenas, Violeta Salteniene, Juozas Kupcinskas, Paulius Matusevicius, Vilmante Borutaite, Limas Kupcinskas |
Journal | World journal of gastroenterology
(World J Gastroenterol)
Vol. 20
Issue 35
Pg. 12533-41
(Sep 21 2014)
ISSN: 2219-2840 [Electronic] United States |
PMID | 25253955
(Publication Type: Journal Article)
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Chemical References |
- Acetophenones
- Anti-Inflammatory Agents
- Enzyme Inhibitors
- Inflammation Mediators
- Tumor Necrosis Factor-alpha
- Dextran Sulfate
- acetovanillone
- Hydrogen Peroxide
- NADH, NADPH Oxidoreductases
- NADPH Oxidase 1
- NADPH Oxidases
- NOX1 protein, mouse
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Topics |
- Acetophenones
(pharmacology)
- Acute Disease
- Animals
- Anti-Inflammatory Agents
(pharmacology)
- Cells, Cultured
- Chronic Disease
- Colitis
(chemically induced, enzymology, pathology, prevention & control)
- Colon
(drug effects, enzymology, pathology)
- Dextran Sulfate
- Disease Models, Animal
- Enzyme Inhibitors
(pharmacology)
- Epithelial Cells
(drug effects, enzymology, pathology)
- Hydrogen Peroxide
(metabolism)
- Inflammation Mediators
(metabolism)
- Intestinal Mucosa
(drug effects, enzymology, pathology)
- Male
- Mice, Inbred BALB C
- NADH, NADPH Oxidoreductases
(antagonists & inhibitors, metabolism)
- NADPH Oxidase 1
- NADPH Oxidases
(antagonists & inhibitors, metabolism)
- Signal Transduction
(drug effects)
- Tumor Necrosis Factor-alpha
(metabolism)
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