Effect of Silymarin on Cathepsin Activity and Oxidative Stress in TNBS-induced Colitis in Rats

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Gokhan Bayramoglu
Hakan Senturk
Gungor Kanbak
Mediha Canbek
Aysegul Bayramoglu
Eda Dokumacioglu
Selin Engur


The purpose of the this study was to defined protective effects of silymarin on the experimental colitis induced by intra-colonic application of 2,4,6-trinitrobenzene sulfonic acid (TNBS).  The twenty eight Sprague-Dawley rats were randomly seperated into four groups, each group containing seven rats represent as follows: group 1 determined as control; group 2 determined as colitis-untreated; group 3 determined as colitis rats administered silymarin (50 mg/kg) and group 4 determined as colitis rats administered silymarin (100 mg/kg). Doses of 50 mg/kg and 100 mg/kg silymarin decreased tissue levels of malondialdehyde (MDA), cathepsin L and cathepsin B and activity of myleperoxidase (MPO) enzyme with respect to the colitis group (p<0.05). Based on the results of the study, it can be said that silymarin can be used as an effective treatment agent in inflammatory bowel diseases.

Cathepsin, colitis, myleperoxidase, oxidative stress, silymarin.

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How to Cite
Bayramoglu, G., Senturk, H., Kanbak, G., Canbek, M., Bayramoglu, A., Dokumacioglu, E., & Engur, S. (2019). Effect of Silymarin on Cathepsin Activity and Oxidative Stress in TNBS-induced Colitis in Rats. Asian Journal of Biochemistry, Genetics and Molecular Biology, 2(4), 1-9. https://doi.org/10.9734/ajbgmb/2019/v2i430072
Original Research Article


Adams SM, Bornemann PH. Ulcerative colitis. Am Fam Physician. 2013;87(10): 699-705.

Galvez J, de Souza Gracioso J, Camuesco D, Galvez J, Vilegas W, Monteiro Souza Brito AR, Zarzuelo A. Intestinal antiinflammatory activity of a lyophilized infusion of Turnera ulmifolia in TNBS rat colitis. Fitoterapia. 2006;77(7-8):515-20.

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(3):483-91.

Zerin M, Karakilcik AZ, Bitiren M, Musa D, Ozgonul A, Selek S, Nazligul Y, Uzunköy A. Vitamin C modulates oxidative stress-induced colitis in rats. Turk J Med Sci. 2010;40(6):871-879.

Pravda J. Radical induction theory of ulcerative colitis. World J Gastroenterol. 2005;11:2371-84.

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;103:177-185.

Wirtz S, Popp V, Kindermann M, Gerlach K, Weigmann B, Fichtner-Feigl S, Neurath MF. Chemically induced mouse models of intestinal inflammation. Nat Protoc. 2007;2: 541-46.

Morris GP, Beck PL, Herridge MS, Depew WT, Szewczuk MR, Wallace JL. Hapten-induced model of chronic inflammation and ulceration in the rat colon. Gastroenterology. 1989;96:795-803.

Sener G, Aksoy H, Sehirli O, Yüksel M, Aral C, Gedik N, Cetinel S, Yeğen BC. Erdosteine prevents colonic inflammation through its antioxidant and free radical scavenging activities. Dig Dis Sci. 2007;52: 2122-32.

Niki E. Lipid peroxidation products as oxidative stress biomarkers. Biofactors. 2008;34(2):171-80.

Pisoschi AM, Pop A. The role of antioxidants in the chemistry of oxidative stress: A review. Eur J Med Chem. 2015; 97:55-74.

Klebanoff SJ. Myeloperoxidase: Friend and foe. J Leukoc Biol. 2005;77:598-625.

Lavelli V, Peri C, Rizzola A. Antioxidant activity of tomato products as studied by model reactions using xanthine oxidase, myeloperoxidase, and copperinduced lipid peroxidation. J Agric Food Chem. 2000;48 (5):1442-1448.

Chapman AL, Mocatta TJ, Shiva S, Seidel A, Chen B, Khalilova I, Paumann-Page ME, Jameson GN, Winterbourn CC, Kettle AJ. Ceruloplasmin is an endogenous inhibitor of myeloperoxidase. J Biol Chem. 2013;288(9):6465-77.

Zavosnik-Bergant T, Turk B. Cysteine cathepsins in the immune response. Tissue Antigens. 2006;67(5):349-55.

Vasiljeva O, Reinheckel T, Peters C, Turk D, Turk V, Turk B. Emerging roles of cysteine cathepsins in disease and their potential as drug targets. Curr Pharm Des. 2007;13:387-403.

Turk V, Turk B, Turk D.Lysosomal cysteine proteases: Facts and opportunities. EMBOJ. 2001;20(17):4629-33.

Brix K, Dunkhorst A, Mayer K, Jordans S. Cysteine cathepsins: Cellular roapmap to different functions. Biochimie. 2008;90(2): 194-207.

Berdowska I. Cysteine proteses as disease markers. Clin Chim Acta. 2004;342(1-2): 41-69.

Siczek K, Zatorski H, Pawlak W, Fichna J. Rhenium-coated glass beads for intracolonic administration attenuate TNBS-induced colitis in mice: Proof-of-Concept Study. Folia Med Cracov. 2015; 55(4):49-57.

Xie Y, Zhang D, Zhang J, Yuan J. Metabolism, Transport and Drug-Drug Interactions of Silymarin. Molecules. 2019; 24(20).

Polyak SJ, Ferenci P, Pawlotsky JM. Hepatoprotective and Antiviral Functions of Silymarin Components in HCV Infection. Hepatology. 2013;57(3):1262–1271.

Abenavoli L, Izzo AA, Milić N, Cicala C, Santini A, Capasso R. Milk thistle (Silybum marianum): A concise overview on its chemistry, pharmacological and nutraceutical uses in liver diseases. Phytother Res. 2018;32(11):2202-2213.

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(13): 146-52.

Wang L, Huang QH, Li YX, Huang YF, Xie JH, Xu LQ, Dou YX, Su ZR, Zeng HF, Chen JN. Protective effects of silymarin on triptolide-induced acute hepatotoxicity in rats. Mol Med Rep. 2018;17(1):789-800.

Ohkawa H, Oshishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbuturic acid reaction. Anal Biochem. 1979;95:351-358.

Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem. 1976; 72:248-254.

Bradley PP, Preibat D, Christerser RD, Rothstein G. Measurement of cutaneous inflammation: Estimation of neutrophil content with an enzyme marker. J Invest Dermatol. 1982;78:206-209.

Barrett AJ, Kirschke H. Methods Enzymol. 1981;80:535-538.

Hanauer SB. Inflammatory bowel disease: Epidemiology, pathogenesis, and therapeutic opportunities. Inflamm Bowel Dis. 2006;12:3-9.

Dijkstra B, Bagshaw PF, Frizelle FA. Protective effect of appendectomy against the development of ulcerative colitis: matched, case-control study. Dis Colon Rectum. 1999;42(3):334-6.

Bernstein CN. Assessing environmental risk factors affecting the inflammatory bowel diseases: A joint workshop of the Crohn's & Colitis Foundations of Canada and the USA. Inflamm Bowel Dis. 2008;14: 1139-46.

Rana SV, Sharma S, Prasad KK, Sinha SK. Singh K. Role of oxidative stress & antioxidant defence in ulcerative colitis patients from north India. Indian J Med Res. 2014;139(4):568-71.

Freeman BA, Crago JD. Biology of the disease free radicals and tissue injury. Lab Invest. 1982;47:412-26.

Drapper H, Hadley M. Malondialdehyde determination as index of lipit peroxidation. Methods Enzymol. 1990;186: 421-431.

Menekse E, Aydın S, Sepici Dinçel A, Eroğlu A, Dolapçı M, Yıldırım O, Cengiz Ö. Effect of 3-aminobenzamide on perforation an experimental colitis model. Turk J Gastroenterol. 2014;25(1):86-91.

Lee IA, Hyun YJ, Kim DH. Berberine ameliorates TNBS-induced colitis by inhibiting lipid peroxidation, enterobacterial growth and NF-κB activation. Eur J Pharmacol. 2010;648(1-3):162-70.

Nencini C, Giorgi G, Micheli L. Protective effect of silymarin on oxidative stress in rat brain. Phytomedicine. 2007;14(2-3):129–135.

Jacobs BP, Dennehy C, Ramirez G, Sapp J, Lawrence VA. Milk thistle for the treatment of liver disease: a systematic review and meta-analysis. Am J Med. 2002;113(6):506-15.

Nieto N, Torres MI, Fernández MI, Girón MD, Ríos A, Suárez MD, Gil A. Experimental Ulcerative Colitis Impairs Antioxidant Defense System in Rat Intestine. Dig Dis Sci. 2000;45(9):1820-7.

Maruyama Y, Lindholm B, Stenvinkel P. Inflammation and oxidative stress in ESRD-the role of myeloperoxidase. J Nephrol 2004;17(8):72-6.

Raab Y, Hällgren R, Knutson L, Krog M, Gerdin B. A technique for segmental rectal and colonic perfusion in humans. Am J Gastroenterol. 1992;87(10):1453-9.

Masoodi I, Tijjani BM, Wani H, Hassan NS, Khan AB, Hussain S. Biomarkers in the management of ulcerative colitis: A brief review. Ger Med Sci. 2011;9:Doc03.

Videla S, Vilaseca J, Medina C, Mourelle M, Guarner F, Salas A, Malagelada JR. Selective Inhibition of Phosphodiesterase-4 Ameliorates Chronic Colitis and Prevents Intestinal Fibrosis. J Pharmacol Exp Ther. 2006;316(2) 940-5.

Jahovic N, Ercan F, Gedik N, Yüksel M, Sener G, Alican I. The effect of angiotensin-converting enzyme inhibitors on experimental colitis in rats. Regul Pept. 2005;130(1-2):67-74.

Vaziri ND. Oxidative stres in uremia: Nature, mechanisms and potential cosequences. Semin Nephrol. 2004;24(5): 469-73.

Miroliaee AE, Esmaily H, Vaziri-Bami A, Baeeri M, Shahverdi AR, Abdollahi M. Amelioration of experimental colitis by a novel nanoselenium-silymarin mixture. Toxicol Mech Methods. 2011;21(3):200-8.

Esmaily H, Hosseini-Tabatabaei A, Rahimian R, Khorasani R, Baeeri M, Barazesh-Morgani A, Yasa N, Khademi Y, Abdollahi M. On the benefits of silymarin in murine colitis by improving balance of destructive cytokines and reduction of toxic stress in the bowel cells. Cent Eur J Biol. 2009;4:204–213.

Shahbazi F, Khavidaki DS, Khalili H, Pezeshki LM. Potential Renoprotective Effects of Silymarin Against Nephrotoxic Drugs: A Review of Literature. J Pharm Pharmaceut. 2012;15(1):112-123.

Burczynski FJ, Wang G, Nguyen D, Chen Y, Smith HJ, Gong Y. Silymarin and hepatoprotection. Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2012;37(1):6-10.

Stoka V, Turk V, Turk B. Lysosomal cysteine cathepsins: signaling pathways in apoptosis. Biol Chem. 2007;388:555–60.

Ivanova S, Repnik U, Bojic L, Petelin A, Turk V, Turk B. Lysosomes in apoptosis. Methods Enzymol. 2008;442:183-99.

Gondi CS, Rao JS. Cathepsin B as a Cancer Target. Expert Opin Ther Targets. 2013;17(3):281–291.

McGrath ME. The lysosomal cysteine proteases. Annu Rev Biophys Biomol Struct. 1999;28:181–204.

Menzel K, Hausmann M, Obermeier F, Schreiter K, Dunger N, Bataille F, Falk W, Scholmerich J, Herfarth H, Rogler G. Cathepsins B, L and D in inflammatory bowel disease macrophages and potential therapeutic effects of cathepsin inhibition in vivo. Clin Exp Immunol. 2006;146(1):169-80.

Cattaruzza F, Lyo V, Jones E, Pham D, Hawkins J, Kirkwood K, Valdez-Morales E, Ibeakanma C, Vanner SJ, Bogyo M, Bunnett NW. Cathepsin S is activated during colitis and causes visceral hyperalgesia by a PAR2-dependent mechanism in mice. Gastroenterology. 2011;141(5):1864-74.

Moura FA, de Andrade KQ, Dos Santos JCF, Araújo ORP, Goulart MOF. Antioxidant therapy for treatment of inflammatory bowel disease: Does it work? Redox Biol. 2015;6:617-39.