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Investigating Intestinal Inflammation in DSS-induced Model of IBD
Published on: February 1, 2012
Evidence for oxidative stress in NSAID-induced colitis in IL10-/- mice
Seiko Narushima1, Douglas R Spitz, Larry W Oberley
1Department of Internal Medicine, University of Iowa College of Medicine, Iowa City 52242, USA.
Abstract:
The goal of this study was to evaluate for evidence of oxidative stress in colonic inflammation in a novel model of inflammatory bowel disease, nonsteroidal anti-inflammatory drug- (NSAID-) treated interleukin-10-deficient (IL10(-/-)) mice. IL10(-/-) and wild-type (wt) mice were treated with a nonselective NSAID (piroxicam, 200 ppm in the diet) for 2 weeks to induce colitis, and parameters for oxidative stress in the colonic tissues were evaluated. Mean chemiluminescence enhanced with lucigenin in the colons from IL10(-/-) mice treated with piroxicam was more than 5-fold higher than that of the control wt group. Chemiluminescence was inhibited with diphenylethylene iodinium, but not allopurinol, indomethacin, or N-omega-nitro-L-arginine, indicating that flavin-containing enzymes were the source of the reactive oxygen species. Colonic aconitase activity in NSAID-treated IL10(-/-) mice decreased to 50% of the activity of control mice. There was no difference in the total glutathione levels in the colonic mucosa among the groups; however, glutathione disulfide levels were approximately 2-fold greater in the colon of NSAID-treated IL10(-/-) mice as compared with control groups. Immunohistochemistry studies of colons from NSAID-treated IL10(-/-) mice demonstrated intense staining with two antibodies that recognize advanced glycation endproducts formed through glycation and oxidation: anticarboxymethylysine and antipentosidine. The epithelial cells and lamina propria cells in the colons of NSAID-treated IL10(-/-) mice showed immunostaining with antinitrotyrosine, indicating the presence of reactive nitrogen species. Colonic epithelium of IL10(-/-) mice with colitis showed moderate immunostaining for 8-hydroxy-2'-deoxyguanosine in the nuclei. NSAID-treated IL10(-/-) mice treated with diphenylene idodonium chloride (DPI), an irreversible inhibitor of flavoprotein enzymes, experienced significantly reduced inflammation. Taken together, these results strongly indicate the presence of oxidative stress in the inflammatory bowel disease in NSAID-treated IL10(-/-) mice and suggests a role for oxidative stress in the pathophysiology of this model of inflammatory bowel disease.
Insights
This study shows oxidative stress in a novel inflammatory bowel disease model using NSAID-treated IL10(-/-) mice. Evidence suggests reactive oxygen and nitrogen species contribute to the disease, with flavin-containing enzymes being a key source.
Area of Science:
- Gastroenterology
- Immunology
- Biochemistry
Background:
- Inflammatory bowel disease (IBD) pathogenesis is complex.
- Oxidative stress is implicated in various inflammatory conditions.
- Interleukin-10 deficient (IL10(-/-)) mice are a model for IBD, but NSAID-induced colitis offers a novel approach.
Purpose of the Study:
- To investigate oxidative stress markers in NSAID-induced colitis in IL10(-/-) mice.
- To identify the source of reactive oxygen species (ROS) in this model.
- To explore the role of oxidative and nitrosative stress in IBD pathophysiology.
Main Methods:
- Induction of colitis in IL10(-/-) and wild-type mice using piroxicam (a non-selective NSAID).
- Measurement of chemiluminescence, enzyme activities (aconitase), and glutathione levels in colonic tissues.
- Immunohistochemical analysis for advanced glycation endproducts, nitrotyrosine, and 8-hydroxy-2'-deoxyguanosine.
- Inhibition studies using diphenylene iodonium chloride (DPI).
Main Results:
- NSAID-treated IL10(-/-) mice exhibited significantly increased colonic chemiluminescence, indicating heightened ROS production.
- ROS production was linked to flavin-containing enzymes, as shown by inhibition with diphenylene iodonium chloride.
- Decreased aconitase activity and increased glutathione disulfide levels suggested oxidative damage.
- Immunohistochemistry revealed the presence of advanced glycation endproducts, nitrotyrosine, and 8-hydroxy-2'-deoxyguanosine, indicating oxidative and nitrosative stress.
- DPI treatment significantly reduced inflammation in NSAID-treated IL10(-/-) mice.
Conclusions:
- NSAID-induced colitis in IL10(-/-) mice presents significant evidence of oxidative and nitrosative stress.
- Flavin-containing enzymes are a likely source of ROS contributing to inflammation in this IBD model.
- These findings highlight the critical role of oxidative stress in the pathophysiology of this novel IBD model.
