The effect of betulinic acid on TNBS-induced experimental colitis

Abstract

ABSTRACT: In this study we have investigated the possible protective effect of betulinic acid
(BA) on colonic inflammation in rats. Colitis was induced in Sprague-Dawley rats of both
sexes by intracolonic administration of 1 ml trinitrobenzene sulphonic acid (TNBS). Colitisinduced
rats received orogastrically either betulinic acid (50 mg/kg/day) or vehicle (0.05%
DMSO) for 3 days. At the 72nd hour of colitis induction, the rats were decapitated and trunk
blood was collected for the measurement of TNF-, IL-1, lactate dehydrogenase (LDH) levels
and total antioxidant capacity (AOC). The distal 8 cm of colon were scored macroscopically,
and the degree of oxidant damage was evaluated by malondialdehyde (MDA) and
glutathione (GSH) levels, myeloperoxidase activity (MPO), collagen content and by histological
analysis. Generation of oxidants was evaluated by tissue luminol and lucigenin
chemiluminescences (CL). Colitis caused significant increases in the colonic CL values,
macroscopic damage scores, MDA, MPO and collagen levels, along with a significant decrease
in tissue GSH level. Similarly, serum TNF-, IL-1, as well as LDH were elevated and
AOC was reduced in the vehicle-treated colitis group as compared to control group. On the
other hand, betulinic acid treatment reversed all these biochemical indices, as well as histopathological
alterations induced by TNBS, suggesting that betulinic acid protects the colonic
tissue via its radical scavenging and antioxidant activities.
KEY WORDS: betulinic acid; colitis; oxidative damage; inflammation;
trinitrobenzene sulphonic acid

Keywords

• betulinic acid; colitis; oxidative damage; inflammation; trinitrobenzene sulphonic acid

The effect of betulinic acid on TNBS-induced experimental colitis

Abstract

References

1. Shanahan F. Pathogenesis of ulcerative colitis. Lancet 1993;342:407–11.
2. Keshavarzian A, Sedghi S, Kanofsky J, List T, Robinson C, Ibrahim C, Winship D. Excessive production of reactive oxygen metabolites by inflamed colon: analysis by chemiluminescence probe. Gastroenterology 1992;103: 177-85.
3. Podolsky DK. Inflammatory bowel disease. N Engl J Med 2002;347:417-29.
4. Ghosh P, Mandal A, Chakraborty P, Rasul MG, Chakraborty M, Saha A. Triterpenoids from Psidium guajava with Biocidal Activity. Indian J Pharm Sci 2010;72:504-7.
5. Isik F, Tunali Akbay T, Yarat A, Genc Z, Pisiriciler R, Caliskan-Ak E, Cetinel S, Altıntas A, Sener G. Protective Effects of Black Cumin (Nigella sativa) Oil on TNBS-Induced Experimental Colitis in Rats. Dig Dis Sci 2011;256:721-30.
6. Genc Z, Yarat A, Tunali-Akbay T, Sener G, Cetinel S, Pisiriciler R, Caliskan-Ak E, Altıntas A, Demirci B. The Effect of Stinging Nettle (Urtica dioica) Seed Oil on Experimental Colitis in Rats. J Med Food 2011; 14:1554-61.
7. Sami A, Taru MK, Salme K, Jari YK. Pharmacological properties of the ubiquitous natural product betulin. Eur J Pharm Sci 2006;29:1-13.
8. Aiken C, Chen CH. Betulinic acid derivatives as HIV-1 antivirals. Trends Mol Med 2005;11: 31-6.

9. Ekşioğlu-Demiralp E, Kardaş ER, Ozgül S, Yağcı T, Bilgin H, Sehirli O, Ercan F, Sener G. Betulinic acid protects against ischemia/reperfusion-induced renal damage and inhibits leukocyte apoptosis. Phytother Res 2010; 24:325-32.
10. Wallace JL, Braquet P, Ibbotson GC, MacNaugton WK, Cirino G. Assessment of the role of platelet activating factor in an animal model of inflammatory bowel disease. J Lipid Med 1989;1: 13-23.
11. Martinek RG. A rapid ultraviolent spectrophotomeetric lactic dehydrogenase assay. Clin Chem Acta 1972;40:91-9.
12. Haklar G, Ulukaya-Durakbaşa C, Yüksel M, Daglı T, Yalcin AS. Oxygen radicals and nitric oxide in rat mesenteric ischemia-reperfusion: modulation by L-arginine and N-nitro-L-arginine methyl ester. Clin Exp Pharmacol Physiol 1998;25: 908-12.
13. Beuge JA, Aust SD. Microsomal lipid peroxidation. Meth Enzymol 1978;52:302-11.
14. Beutler E. Glutathione in red blood cell metabolism. A Manuel of Biochemical Methods, Grune&Stratton, New York 1975,112-114.
15. Hillegass LM, Griswold DE, Brickson B, AlbrightsonWinslow C. Assessment of myeloperoxidase activity in whole rat kidney. J Pharmacol Meth 1990; 24:285-295.
16. Bradley PP, Priebat DA, Christersen RD, Rothstein G. Measurement of cutaneous inflammation. Estimation of neutrophil content with an enzyme marker. J Invest Dermatol 1982;78:206–9.
17. Lopez De Leon A, Rojkind M. A simple micromethod for collagen and total protein determination in formalinfixed parraffin-embedded sections. J Histochem Cytochem 1985;33: 737-43.
18. O’Shea JJ, Murray PJ. Cytokine signaling modules in inflammatory responses. Immunity 2008;28:477–87.
19. Amini-Shirazi N, Hoseini A, Ranjbar A, Mohammadirad A, Khoshakhlagh P, Yasa N, Abdollahi M. Inhibition of tumor necrosis factor and nitrosative/oxidative stresses by Ziziphora clinopoides (Kahlioti); a molecular mechanism of protection against dextran sodium sulfate-induced colitis in mice. Toxicol Mech Methods 2009;19:183–9.
20. Cetinel S, Hancioğlu S, Sener E, Uner C, Kiliç M, Sener G, Yeğen BC . Oxytocin treatment alleviates stress-aggravated colitis by a receptor-dependent mechanism. Regul Pept 2010;160:146-52.
21. Emekli-Alturfan E, Yarat A, Tunali-Akbay T, Isik F, Yenidogan G, Sener G, Sehirli O, Pisiriciler R, Ak E, Altintas A. Effect of Black Cumin (Nigella Sativa) Seed Oil on Gastric Tissue in Experimental Colitis. Adv Environ Biol 2011;5:483-90.
22. Şehirli AÖ, Tatlıdede E, Yüksel M, Çetinel Ş, Erzik C, Yeğen B, Şener G. Protective effects of alpha-lipoic acid against oxidative injury in TNBS-induced colitis. Erciyes Med J 2009;31:15-26.
23. Ross D. Glutathione, free radicals and chemotherapeutic agents. Mechanisms of free-radical induced toxicity and glutathione-dependent protection. Pharmacol Ther 1988;37:231–49.
24. Reiter RJ, Tan DX, Osuna C, Gitto E. Actions of melatonin in the reduction of oxidative stress. A review. J Biomed Sci 2000;7:444–58.
25. Elson CO, Sartor RB, Tennyson GS, Riddell RH. Experimental models of inflammatory bowel disease. Gastroenterology 1995;109:1344–67.
26. Işeri SO, Sener G, Sağlam B, Gedik N, Ercan F, Yeğen BC. Oxytocin ameliorates oxidative colonic inflammation by a neutrophil-dependent mechanism. Peptides. 2005;26:483-91.
27. Jahovic N, Gedik N, Ercan F, Sirvanci S, Yüksel M, Sener G, Alican I.Effects of statins on experimental colitis in normocholesterolemic rats. Scand J Gastroenterol. 2006;41:954-62.