Korean J Gastroenterol.  2013 Jul;62(1):33-41. 10.4166/kjg.2013.62.1.33.

Benzoxazole Derivative B-98 Ameliorates Dextran Sulfate Sodium-induced Acute Murine Colitis and the Change of T Cell Profiles in Acute Murine Colitis Model

• Song EM
• Jung SA
• Lee JS
• Kim SE
• Shim KN
• Jung HK
• Yoo K
• Park HY

• KMID: 1711277
• DOI: http://doi.org/10.4166/kjg.2013.62.1.33

Abstract

BACKGROUND/AIMS
The unique role of enzyme 5-lipoxygenase (5-LO) in the production of leukotrienes makes it a therapeutic target for inflammatory bowel disease (IBD). The aim of this study was to evaluate the effects of B-98, a newly synthesized benzoxazole derivatives and a novel 5-LO inhibitor, in a mouse model of IBD induced by dextran sulfate sodium (DSS).
METHODS
C57BL/6 mice were randomly assigned to four groups: normal control, DSS colitis (DSS+saline), low dose B-98 (DSS+B-98 20 mg/kg) and high dose B-98 (DSS+B-98 100 mg/kg). B-98 was administered with 3% DSS intraperitoneally. The severity of the colitis was assessed via the disease activity index (DAI), colon length, and histopathologic grading. The production of inflammatory cytokines interleukin (IL)-6 was determined by RT-PCR. Th cells were examined for the proportion of Th1 cell, Th2 cell, Th9 cell, Th17 cell and Treg cell using intracellular cytometry.
RESULTS
The B-98 group showed lower DAI, less shortening of the colon length and lower histopathologic grading compared with the DSS colitis group (p<0.01). The expression of IL-6 in colonic tissue was significantly lower in the B-98 groups than the DSS colitis group (p<0.05). The cellular profiles revealed that the Th1, Th9 and Th17 cells were increased in the DSS colitis group compared to the B-98 group (p<0.05).
CONCLUSIONS
Our results suggest that acute intestinal inflammation is reduced in the group treated with B-98 by Th1, Th9 and Th17 involved cellular immunity.

Keyword

5-Lipoxygenase inhibitors; Inflammatory bowel diseases; Colitis

MeSH Terms

Acute Disease
Animals
Arachidonate 5-Lipoxygenase/chemistry/metabolism
Benzoxazoles/chemistry/*pharmacology
Colitis/chemically induced/pathology/*prevention & control
Colon/drug effects/pathology/physiology
Dextran Sulfate/toxicity
Disease Models, Animal
Forkhead Transcription Factors/metabolism
Injections, Intraperitoneal
Interleukin-6/genetics/metabolism
Lipoxygenase Inhibitors/chemistry/*pharmacology
Male
Mice
Mice, Inbred C57BL
Severity of Illness Index
T-Lymphocytes/classification/*drug effects/metabolism
Arachidonate 5-Lipoxygenase
Benzoxazoles
Dextran Sulfate
Forkhead Transcription Factors
Interleukin-6
Lipoxygenase Inhibitors

Figure

• 

  Fig. 1. Chemical structure of B-98 and proposed affinity for the iron of zileuton and B-98. B-98 showed enhanced affinity of 5-lipoxygenase iron than zileuton.

• 

  Fig. 2. Schematic diagram describing type, dose, and timing of each treatment of all four groups. B-98 was injected intraperitoneally from day 1 to day 7. DSS, dextran sulfate sodium; po, per oral; ip, intraperitoneal.

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  Fig. 3. Effect of B-98 on the course of disease activity index (DAI). At day 4 after dextran sulfate sodium (DSS) administration, the gross rectal bleeding appeared. Administration of B-98 significantly attenuated the severity of clinical DAI compared with the corresponding DSS colitis group at day 7 after DSS administration (*p＜0.05). Data are expressed as means.

• 

  Fig. 4. (A) Histological appearance of the colon. a: Colon section from the normal control group; H&E, ×40 and ×100 for a-1 and a-2, respectively. b: Colon section from the dextran sulfate sodium (DSS) colitis group. Extensive erosions with predominantly inflammatory cell infiltrates in mucosa and submucosal layer were evident; H&E, ×40 and ×100 for b-1 and b-2, respectively. c: Colon section from the low dose B-98 (20 mg/kg) group. The mucosa was essentially unremarkable. There were fewer inflammatory cell infiltrates, erosions, or crypt distortion compared with b; H&E, ×40 and ×100 for c-1 and c-2, respectively. d: Colon section from the high dose B-98 (100 mg/kg) group. There were fewer inflammatory infiltrates, erosions, or crypt distortion compared with b. Occasionally, inflammatory cell infiltrates were also seen in the submucosal layer d-2; H&E, ×40 and ×100 for d-1 and d-2, respectively. (B) Histological scores of the DSS colitis and B-98 groups. Histological score levels were significantly lower in the low dose B-98 group in comparison with the DSS colitis group (*p＜0.05).

• 

  Fig. 5. Expression of inflammatory cytokine. Interleukin-6 (IL-6) concentration of the colonic tissue in the dextran sulfate sodium (DSS) colitis group was higher than that of the normal control group. And IL-6 level in the B-98 groups was significantly lower than that of the DSS colitis group (*p＜0.05). Results are presented as fold- increase relative to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) expression. Bold line in the box means median values.

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  Fig. 6. Flow cytometric characteristics of CD4＋ T cell in the B-98 group. The proportion of CD4＋ T cells from lamina propria were determined according to intracellular levels of cytokines for interferon (IFN)-γ＋ Th1 cell, interleukin (IL)-4＋ Th2 cells, IL-9＋ Th9 cells, IL-17＋ Th17 cells, and Foxp3＋ Treg cells. The numbers of inside quadrant indicate the percentage of the cell population (*p＜0.05). DSS, dextran sulfate sodium.

Reference

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