Inflammatory bowel disease (IBD) is a chronic inflammation of the GI tract that includes Crohn's disease and ulcerative colitis. The major enzymes involved in mediating the inflammatory response in IBD are cyclooxygenase (COX), lipoxygenase (LOX) and nitric oxide synthase (NOS). The exact mechanisms by which these enzymes are induced and inflammation is caused are still not fully understood. The objective of this proposal is to develop in vivo microdialysis sampling in the colonic mucosa to be used in conjunction with microanalytical techniques to monitor the activity of these three enzyme systems in an animal model of IBD. Three analytical methods will be developed as part of this project. First, the COX products prostaglandin E2 and thromboxane B2 and the LOX products the hydroxyeicosatetraenoic acids, 5-HETE and 15-HETE, and leukotriene B4 will be determined by CE-LIF after derivitization with either 9- anthryldiazomethane (ADAM) or 2-(2, 3-naphthalimino) ethyltrifluoromethanesulphonate (NE-OTf). Second, the NOS and arginase products citrulline and ornithine will be determined using CE-LIF after derivatization by naphthalene-2,3-dicarboxaldehyde/cyanide (NDA/CN). Third, nitrate and nitrite will be used to monitor NOS activity and determined directly by CE-UV or by CE-LIF following derivatization with 2,3-diaminoaphtalene (DAN). The animal model of IBD to be used will be by chemical induction using 2,4,6-trinitrobenzene sulfonic acid in the rat. Implementation of this model will involve development of microdialysis probe implantation protocols, evaluation of any tissue response due to probe implantation, and verification that the probe is located in the inflamed region. Microdialysis sampling and the developed analytical methods will then be used to continuously monitor the enzyme activity in the model as the animal develops IBD and in control animals.

Public Health Relevance

Statement Inflammatory bowel disease (IBD) affects 1.4 million people in the United States and 2.2 million people in Europe. The development of fully effective treatments is hindered by the complexity of the enzyme systems involved in the inflammatory response. The goal of this project is to develop analytical and in vivo sampling methods to investigate the interactions of the enzyme systems implicated in IBD.

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Research Project (R01)
Project #
2R01EB000247-18A2
Application #
7731772
Study Section
Enabling Bioanalytical and Biophysical Technologies Study Section (EBT)
Program Officer
Korte, Brenda
Project Start
1991-06-01
Project End
2011-06-30
Budget Start
2009-07-15
Budget End
2010-06-30
Support Year
18
Fiscal Year
2009
Total Cost
$333,666
Indirect Cost
Name
University of Kansas Lawrence
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
076248616
City
Lawrence
State
KS
Country
United States
Zip Code
66045
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Hoque, Mohammed E; Arnett, Stacy D; Lunte, Craig E (2005) On-column preconcentration of glutathione and glutathione disulfide using pH-mediated base stacking for the analysis of microdialysis samples by capillary electrophoresis. J Chromatogr B Analyt Technol Biomed Life Sci 827:51-7

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