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

Affiliations
  • 1Department of Internal Medicine, Ewha Womans University School of Medicine, Ewha Womans University, Seoul, Korea. jassa@ewha.ac.kr
  • 2College of Pharmacy, Ewha Womans University, Seoul, Korea.

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.

  • 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.

  • 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.


Cited by  1 articles

Anti-inflammatory effects of DA-9601, an extract of Artemisia asiatica, on aceclofenac-induced acute enteritis
Ju Hwan Kim, Chang Yell Shin, Sun Woo Jang, Dong-Seok Kim, Wonae Lee, Hyung-Gun Kim, Hak Rim Kim
Korean J Physiol Pharmacol. 2021;25(5):439-448.    doi: 10.4196/kjpp.2021.25.5.439.


Reference

References

1. Yang SK, Yun S, Kim JH, et al. Epidemiology of inflammatory bowel disease in the Songpa-Kangdong district, Seoul, Korea, 1986–2005: a KASID study. Inflamm Bowel Dis. 2008; 14:542–549.
Article
2. Podolsky DK. Inflammatory bowel disease. N Engl J Med. 2002; 347:417–429.
Article
3. Neurath MF, Finotto S, Glimcher LH. The role of Th1/Th2 polar-ization in mucosal immunity. Nat Med. 2002; 8:567–573.
Article
4. Duerr RH, Taylor KD, Brant SR, et al. A genome-wide association study identifies IL23R as an inflammatory bowel disease gene. Science. 2006; 314:1461–1463.
5. Mudter J, Neurath MF. Il-6 signaling in inflammatory bowel disease: pathophysiological role and clinical relevance. Inflamm Bowel Dis. 2007; 13:1016–1023.
Article
6. Olsen T, Rismo R, Cui G, Goll R, Christiansen I, Florholmen J. TH1 and TH17 interactions in untreated inflamed mucosa of inflammatory bowel disease, and their potential to mediate the inflammation. Cytokine. 2011; 56:633–640.
Article
7. Sakaguchi S, Sakaguchi N, Asano M, Itoh M, Toda M. Immunologic self-tolerance maintained by activated T cells expressing IL-2 receptor alpha-chains (CD25). Breakdown of a single mechanism of self-tolerance causes various autoimmune diseases. J Immunol. 1995; 155:1151–1164.
8. Henderson WR Jr. The role of leukotrienes in inflammation. Ann Intern Med. 1994; 121:684–697.
Article
9. Dahlén SE, Björk J, Hedqvist P, et al. Leukotrienes promote plasma leakage and leukocyte adhesion in postcapillary venules: in vivo effects with relevance to the acute inflammatory response. Proc Natl Acad Sci U S A. 1981; 78:3887–3891.
10. Rachmilewitz D, Simon PL, Schwartz LW, Griswold DE, Fonda-caro JD, Wasserman MA. Inflammatory mediators of experimental colitis in rats. Gastroenterology. 1989; 97:326–337.
Article
11. Boughton-Smith NK, Hawkey CJ, Whittle BJ. Biosynthesis of lip-oxygenase and cyclooxygenase products from [14C]-arachi-donic acid by human colonic mucosa. Gut. 1983; 24:1176–1182.
Article
12. Berger W, De Chandt MT, Cairns CB. Zileuton: clinical implications of 5-Lipoxygenase inhibition in severe airway disease. Int J Clin Pract. 2007; 61:663–676.
Article
13. Song H, Oh SR, Lee HK, et al. Synthesis and evaluation of benzoxazole derivatives as 5-lipoxygenase inhibitors. Bioorg Med Chem. 2010; 18:7580–7585.
Article
14. Stevceva L, Pavli P, Husband A, Ramsay A, Doe WF. Dextran sulphate sodium-induced colitis is ameliorated in interleukin 4 deficient mice. Genes Immun. 2001; 2:309–316.
Article
15. Kihara N, de la Fuente SG, Fujino K, Takahashi T, Pappas TN, Mantyh CR. Vanilloid receptor-1 containing primary sensory neurones mediate dextran sulphate sodium induced colitis in rats. Gut. 2003; 52:713–719.
Article
16. Jang MH, Sougawa N, Tanaka T, et al. CCR7 is critically important for migration of dendritic cells in intestinal lamina propria to mesenteric lymph nodes. J Immunol. 2006; 176:803–810.
Article
17. Lauritsen K, Laursen LS, Bukhave K, Rask-Madsen J. Effects of topical 5-aminosalicylic acid and prednisolone on prostaglandin E2 and leukotriene B4 levels determined by equilibrium in vivo dialysis of rectum in relapsing ulcerative colitis. Gastroenterology. 1986; 91:837–844.
Article
18. Lauritsen K, Laursen LS, Bukhave K, Rask-Madsen J. In vivo effects of orally administered prednisolone on prostaglandin and leucotriene production in ulcerative colitis. Gut. 1987; 28:1095–1099.
Article
19. Cuzzocrea S, Rossi A, Mazzon E, et al. 5-Lipoxygenase modulates colitis through the regulation of adhesion molecule expression and neutrophil migration. Lab Invest. 2005; 85:808–822.
Article
20. Bertrán X, Mañé J, Fernández-Bañares F, et al. Intracolonic administration of zileuton, a selective 5-lipoxygenase inhibitor, accelerates healing in a rat model of chronic colitis. Gut. 1996; 38:899–904.
Article
21. Singh VP, Patil CS, Kulkarni SK. Effect of 5-lipoxygenase inhibition on events associated with inflammatory bowel disease in rats. Indian J Exp Biol. 2004; 42:667–673.
22. Zarif A, Eiznhamer D, Callaghan C, Doria MI, Broutman L, Keshavarzian A. The effect of a selective 5-lipoxygenase inhibitor, zileuton, on tissue damage in acute colonic inflammation in rats. Inflammation. 1996; 20:217–227.
Article
23. Hawkey CJ, Dube LM, Rountree LV, Linnen PJ, Lancaster JF. A trial of zileuton versus mesalazine or placebo in the maintenance of remission of ulcerative colitis. The European Zileuton Study Group For Ulcerative Colitis. Gastroenterology. 1997; 112:718–724.
Article
24. Holma R, Salmenperä P, Riutta A, Virtanen I, Korpela R, Vapaatalo H. Acute effects of the cys-leukotriene-1 receptor antagonist, montelukast, on experimental colitis in rats. Eur J Pharmacol. 2001; 429:309–318.
Article
25. Hawthorne AB, Boughton-Smith NK, Whittle BJ, Hawkey CJ. Colorectal leukotriene B4 synthesis in vitro in inflammatory bowel disease: inhibition by the selective 5-lipoxygenase inhibitor BWA4C. Gut. 1992; 33:513–517.
Article
26. Ito R, Kita M, ShinYa M, et al. Involvement of IL-17A in the pathogenesis of DSS-induced colitis in mice. Biochem Biophys Res Commun. 2008; 377:12–16.
Article
27. Leppkes M, Becker C, Ivanov II, et al. RORgamma-expressing Th17 cells induce murine chronic intestinal inflammation via re-dundant effects of IL-17A and IL-17F. Gastroenterology. 2009; 136:257–267.
28. Alex P, Zachos NC, Nguyen T, et al. Distinct cytokine patterns identified from multiplex profiles of murine DSS and TNBS-induced colitis. Inflamm Bowel Dis. 2009; 15:341–352.
Article
29. Stassen M, Schmitt E, Bopp T. From interleukin-9 to T helper 9 cells. Ann N Y Acad Sci. 2012; 1247:56–68.
Article
30. Mottet C, Uhlig HH, Powrie F. Cutting edge: cure of colitis by CD4+CD25+ regulatory T cells. J Immunol. 2003; 170:3939–3943.
Article
Full Text Links
  • KJG
Actions
Cited
CITED
export Copy
Close
Share
  • Twitter
  • Facebook
Similar articles
Copyright © 2024 by Korean Association of Medical Journal Editors. All rights reserved.     E-mail: koreamed@kamje.or.kr