Thymol has beneficial effects on the experimental model of ulcerative colitis

Document Type : Original Research Article


Department of Microbiology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran.


Objective: Thymol, a natural aromatic monoterpene phenol derived from thymus, possesses anti-inflammatory benefits. Here, we evaluated the potential of thymol therapy in improving an animal model of ulcerative colitis.
Materials and Methods: Luminal instillation of acetic acid was used to induce colitis in male Wistar rats. Treatment groups daily received prednisolone (2 mg/kg, orally) or thymol (100 mg/kg, orally) for 10 consecutive days. Then, the rats were euthanized and tissue specimens were collected for evaluation of cyclooxygenase-2 (COX-2) expression by immunohistochemistry. Furthermore, the levels of total protein, nitric oxide, myeloperoxidase, malondialdehyde, IL-1, IL-6, and TNF-α were monitored in colonic homogenates. Eventually, the relative mRNA expression of IκBα and NF-κBp65 was investigated using reverse-transcriptase PCR (RT-PCR) in colonic homogenates.
Results: Both medications could reduce the mortality rate and the clinical scores of ulcerative colitis. The COX-2 expression was significantly reduced in the colons of thymol-treated animals compared to the prednisolone group. Also, the myeloperoxidase activity, nitric oxide level and malondialdehyde intensity were decreased in the colons of thymol-treated animals to a greater extent compared to the prednisolone group. Moreover, the total protein content of guts showed significant increases in the guts of thymol-treated animals in comparison to the prednisolone group. Nonetheless, thymol significantly reduced the levels of IL-6, and IL-1 compared to prednisolone. Both medications caused a significant decrease in the mRNA level of NF-κBp65, though the mRNA level of IκBα did not show significant changes between the groups.
Conclusion: Thymol may be a promising agent to ameliorate ulcerative colitis.


Main Subjects

Abtahi Froushani SM, Mashouri S. 2018. The Beneficial effects of hypiran in ameliorating rat model of ulcerative colitis. Zahedan J Res Med Sci, 20:e58919.
Abtahi Froushani SM, Zarei L, Esmaeili Gouvarchin Ghaleh H, Mansori Motlagh B. 2016. Estragole and methyl-eugenol-free extract of Artemisia dracunculus possesses immunomodulatory effects. Avicenna J Phytomed, 6:526-534.
Ahmad A, Khan A, Akhtar F, Yousuf S, Xess I, Khan LA, Manzoor N. 2011. Fungicidal activity of thymol and carvacrol by disrupting ergosterol biosynthesis and membrane integrity against Candida. Eur J Clin Microbiol Infect Dis, 30:41-50.
Al-Rejaie SS, Abuohashish HM, Al-Enazi MM, Al-Assaf AH, Parmar MY, Ahmed MM .2013a. Protective effect of naringenin on acetic acid-induced ulcerative colitis in rats. WJG, 19:5633-5644.
Al-Rejaie, Salem S, Abdulaziz M. Aleisa, Mohamed M. Sayed-Ahmed, Othman A. AL-Shabanah, Hatem M. Abuohashish, Mohammed M. Ahmed, Khaled A. Al-Hosaini, and Mohamed M. Hafez . 2013b. Protective effect of rutin on the antioxidant genes expression in hypercholestrolemic male Westar rat. BMC Complement Altern Med. 13:136-136.
Auphan N, DiDonato JA, Rosette C, Helmberg A, Karin M. 1995. Immunosuppression by glucocorticoids: inhibition of NF-kappa B activity through induction of I kappa B synthesis. Science , 270:286-290.
Barbosa R, Cruz-Mendes Y, Silva-Alves KS, Ferreira-da-Silva FW, Ribeiro NM, Morais LP, Leal-Cardoso JH. 2017. Effects of Lippia sidoides essential oil, thymol, p-cymene, myrcene and caryophyllene on rat sciatic nerve excitability. B Braz J Med Biol Res, 50:e6351.
Blumberg RS, Saubermann LJ, Strober W .1999. Animal models of mucosal inflammation and their relation to human inflammatory bowel disease. Curr Opin Immunol, 11:648-656
Braga PC, Dal Sasso M, Culici M, Bianchi T, Bordoni L, Marabini L. 2006. Anti-inflammatory activity of thymol: inhibitory effect on the release of human neutrophil elastase, Pharmacology, 77:130-136.
Bryan NS, Grisham MB. 2007. Methods to detect nitric oxide and its metabolites in biological samples. Methods Mol Biol, 43:645-657.
Cho JH. 2008. The genetics and immunopathogenesis of inflammatory bowel disease. Nat Rev Immunol, 8:458-466
Consalvi S, Biava M, Poce G .2015. COX inhibitors: a patent review . 2014. Expert Opin Ther Pat, 25:1357-1371.
D'haens G. 2016. Systematic review: second‐generation vs. conventional corticosteroids for induction of remission in ulcerative colitis. Aliment Pharmacol Ther, 44:1018-1029.
Derry S, Moore RA. 2013. Single dose oral celecoxib for acute postoperative pain in adults. Cochrane Database Syst Rev, 22:CD004233.
Liang D, Li F, Fu Y, Cao Y, Song X, Wang T, Wang W, Guo M, Zhou E, Li D, Yang Z, Zhang N. 2014. Thymol inhibits LPS-stimulated inflammatory response via down-regulation of NF-κB and MAPK signaling pathways in mouse mammary epithelial cells Inflammation. Inflammation, 37:214-222.
Elson CO, Sartor RB, Tennyson GS, Riddell RH. 1995. Experimental models of inflammatory bowel disease. Gastroenterology, 109:1344-1367
Fabia R1, Ar'Rajab A, Johansson ML, Andersson R, Willén R, Jeppsson B, Molin G, Bengmark S. 1993. Impairment of bacterial flora in human ulcerative colitis and experimental colitis in the rat. Digestion, 54:248-255.
Fachini-Queiroz FC, Kummer R, Estevão-Silva CF, Carvalho MD, Cunha JM, Grespan R, Bersani-Amado CA, Cuman RK. .2012. Effects of thymol and carvacrol, constituents of Thymus vulgaris L. essential oil, on the inflammatory response. Evid Based Complement Alternat Med, 2012:657026.
Gao Y , Huang Y, Zhao Y, Hu Y, Li Z, Guo Q, Zhao K, Lu N. 2016. LL202 protects against dextran sulfate sodium-induced experimental colitis in mice by inhibiting MAPK/AP-1 signaling. Oncotarget, 7:63981-63994.
 Gholijani N, Amirghofran Z. 2016. Effects of thymol and carvacrol on T-helper cell subset cytokines and their main transcription factors in ovalbumin-immunized mice. J Immunotoxicol, 13:729-737.
Gholijani N, Gharagozloo M, Farjadian S, Amirghofran Z .2015. Modulatory effects of thymol and carvacrol on inflammatory transcription factors in lipopolysaccharide-treated macrophages. J Immunotoxicol, 13:157-64.
Gorgulu S, Yagci G, Kaymakcioglu N, Ozkara M, Kurt B, Ozcan A, Kaya O, Sadir S, Tufan T .2006. Hyperbaric oxygen enhances the efficiency of 5-aminosalicylic acid in acetic acid–induced colitis in rats. Dig Dis Sci, 51:480-487.
Hauso Ø, Martinsen TC, Waldum H. 2015. 5-Aminosalicylic acid, a specific drug for ulcerative colitis. Scand J Gastroenterol, 50:933-941.
Hawes M, Riddle A, Kirk J, Jergens A, Allenspach K. 2018. Interleukin-1beta expression is increased in the duodenum of dogs with idiopathic inflammatory bowel disease. Vet Rec, 3;183:536.
Hindryckx P, Jairath V, D'Haens G. 2016. Acute severe ulcerative colitis: from pathophysiology to clinical management. Nat Rev Gastroenterol Hepatol, 13:654-664
Huang N, Rizshsky L, Hauck CC, Nikolau BJ, Murphy PA, Birt DF .2012. The inhibition of lipopolysaccharide-induced macrophage inflammation by 4 compounds in Hypericum perforatum extract is partially dependent on the activation of SOCS3. Phytochemistry, 76:106-116 .
Joshi R, Kumar S, Unnikrishnan M, Mukherjee T .2005. Free radical scavenging reactions of sulfasalazine, 5-aminosalicylic acid and sulfapyridine: mechanistic aspects and antioxidant activity. Free Radic Res. 39:1163-1172.
Keshavarzian A, Morgan G, Sedghi S, Gordon J, Doria M. 1990. Role of reactive oxygen metabolites in experimental colitis. Gut, 31:786-790
Kim H-S, Berstad A .1992. Experimental colitis in animal models. Scand J Gastroenterol, 27:529-537
Kolios G, Valatas V, Ward SG. 2004. Nitric oxide in inflammatory bowel disease: a universal messenger in an unsolved puzzle. Immunology, 113:427-437 .
Kondamudi PK, Kovelamudi H, Nayak PG, Rao MC, Shenoy RR. 2015. Curcumin half analog modulates interleukin-6 and tumor necrosis factor-alpha in inflammatory bowel disease. Pharmacogn Mag., 11:S296-S302.
Ribeiro AR, Diniz PB, Pinheiro MS, Albuquerque-Júnior RL, Thomazzi SM. 2016. Gastroprotective effects of thymol on acute and chronic ulcers in rats: the role of prostaglandins, ATP-sensitive K+ channels, and gastric mucus secretion. Chemico-biological interactions, 244:121-8.
Rodrigues de Carvalho L, de Brito TV, Simião da C Júnior J, José Dias Júnior G, de Aguiar Magalhãres D, Guimarães Sousa S, Oliveira Silva R, Rodolfo Pereira da Silva F, Fernando Pereira Vasconcelos D, Maria Costa Véras L, de Sousa de Almeida Leite JR, Santos Martins D, da Silva Martins C, Soares de Oliveira J, Barbosa ALDR. 2018. Epiisopiloturine, an imidazole alkaloid, reverses inflammation and lipid peroxidation parameters in the Crohn disease model induced by trinitrobenzenosulfonic acid in Wistar rats. Biomed Pharmacother, 102:278-285.
Li Z, Zheng J, Zhang N, Li C .2016. Berberine improves airway inflammation and inhibits NF-kappaB signaling pathway in an ovalbumin-induced rat model of asthma. Transl Behav Med, 53:999-1005.
Liang D, Li F, Fu Y, Cao Y, Song X, Wang T, Wang W, Guo M, Zhou E, Li D, Yang Z.  .2014. Thymol inhibits LPS-stimulated inflammatory response via down-regulation of NF-kappaB and MAPK signaling pathways in mouse mammary epithelial cells. Inflammation, 37:214-222 .
Liu B, Li S, Sui X, Guo L, Liu X, Li H, Gao L, Cai S, Li Y, Wang T, Piao X. 2018. Root extract of polygonum cuspidatum siebold and zucc. ameliorates dss-induced ulcerative colitis by affecting nf-kappab signaling pathway in a mouse model via synergistic effects of polydatin, resveratrol, and emodin. Front Pharmacol, 9:347.
Low D, Nguyen DD, Mizoguchi E. 2013. Animal models of ulcerative colitis and their application in drug research. Drug Des Devel Ther, 7:1341-1357.
Marchese A, Orhan IE, Daglia M, Barbieri R, Di Lorenzo A, Nabavi SF, Gortzi O, Izadi M, Nabavi SM. 2016. Antibacterial and antifungal activities of thymol: A brief review of the literature. Food Chem, 210:402-414.
Millar AD, Rampton DS, Chander CL, Claxson AW, Blades S, Coumbe A, Panetta J, Morris CJ, Blake DR. 1996. Evaluating the antioxidant potential of new treatments for inflammatory bowel disease using a rat model of colitis. Gut, 39:407-415
Molodecky NA, Soon S, Rabi DM, Ghali WA, Ferris M, Chernoff G, Benchimol EI, Panaccione R, Ghosh S, Barkema HW, Kaplan GG. 2012. Increasing incidence and prevalence of the inflammatory bowel diseases with time, based on systematic review. Gastroenterology, 142:46-54.
Motlagh BM, Ahangaran NA, Abtahi Froushani SM. 2015. Calcitriol modulates the effects of bone marrow-derived mesenchymal stem cells on macrophage functions. IJBMS, 18:672-676.
Gholijani N, Gharagozloo M, Farjadian S, Amirghofran Z .2016. Modulatory effects of thymol and carvacrol on inflammatory transcription factors. J Immunotoxicol, 13:157-164.
Niu X, Zhang H, Li W, Wang Y, Mu Q, Wang X, He Z, Yao H .2015. Protective effect of cavidine on acetic acid-induced murine colitis via regulating antioxidant, cytokine profile and NF-kappaB signal transduction pathways. Chem Biol Interact, 239:34-45.
Orsonneau JL, Douet P, Massoubre C, Lustenberger P, Bernard S. 1989. An improved pyrogallol red-molybdate method for determining total urinary protein. Clin Chem, 35:2233-2236
Pandey A, Verma S, Kumar VL. 2017. Metformin maintains mucosal integrity in experimental model of colitis by inhibiting oxidative stress and pro-inflammatory signaling. Biomed Pharmacother, 94:1121-1128.
Pulli B, Ali M, Forghani R, Schob S, Hsieh KL, Wojtkiewicz G, Linnoila JJ, Chen JW. 2013. Measuring myeloperoxidase activity in biological samples. PloS one, 8:e67976.
Gupta RA, Motiwala MN, Mahajan UN, Sabre SG.2018. Protective effect of Sesbania grandiflora on acetic acid induced ulcerative. J Ethnopharmacol, 219:222-232
Ribeiro ARS, Diniz PB, Pinheiro MS, Albuquerque-Júnior RL, Thomazzi SM .2016. Gastroprotective effects of thymol on acute and chronic ulcers in rats: The role of prostaglandins, ATP-sensitive K+ channels, and gastric mucus secretion. Chem Biol Interact, 244:121-128
Riella K, Marinho R, Santos J, Pereira-Filho R, Cardoso J, Albuquerque-Junior R, Thomazzi S. 2012. Anti-inflammatory and cicatrizing activities of thymol, a monoterpene of the essential oil from Lippia gracilis, in rodents. J Ethnopharmacol, 143:656-663.
Sanei MH, Hadizadeh F, Adibi P, Alavi SA .2014. Inflammatory cells' role in acetic acid-induced colitis. Ad Biomed Res, 3:193.
Strober W. 1985. Animal models of inflammatory bowel disease—an overview. Dig Dis Sci, 30:3S-10S.
Nakazato T, Sagawa M, Yamato K, Xian M, Yamamoto T, Suematsu M, Ikeda Y, Kizaki M. 2007. Myeloperoxidase is a key regulator of oxidative stress mediated apoptosis in myeloid leukemic cells. Clin Cancer Res, 13:5436-5445
Tahan G, Aytac E, Aytekin H, Gunduz F, Dogusoy G, Aydin S, Tahan V, Uzun H .2011. Vitamin E has a dual effect of anti-inflammatory and antioxidant activities in acetic acid-induced ulcerative colitis in rats. Can J Surg, 54:333-338.
Tang X, Chen S, Wang L. 2011. Purification and identification of carvacrol from the root of Stellera chamaejasme and research on its insecticidal activity. Nat Prod Res, 25:320-325.
Wan L, Meng D, Wang H, Wan S, Jiang S, Huang S, Wei L, Yu P. 2018. Preventive and therapeutic effects of thymol in a lipopolysaccharide-induced acute lung injury in mice model. Inflammation, 41:183-192.
Wang HF, Yih KH, Yang CH, Huang KF .2017. Anti-oxidant activity and major chemical component analyses of twenty-six commercially available essential oils. J Food Drug Anal, 25:881-889 .
Wang Y, Parker CE, Feagan BG, MacDonald JK. 2016. Oral 5‐aminosalicylic acid for maintenance of remission in ulcerative colitis. Cochrane Database Syst Rev., 9:CD000544.
Winterbourn CC, Kettle AJ, Hampton MB. 2016. Reactive oxygen species and neutrophil function. Annual review of biochemistry, 85:765-92.
Wu H, Jiang K, Yin N, Ma X, Zhao G, Qiu C, Deng G. 2017. Thymol mitigates lipopolysaccharide-induced endometritis by regulating the TLR4- and ROS-mediated NF-kappaB signaling pathways. Oncotarget, 8:20042-20055.
Xiao HB, Wang CR, Liu ZK, Wang JY. 2015. LPS induces pro-inflammatory response in mastitis mice and mammary epithelial cells: Possible involvement of NF-κB signaling and OPN. Pathologie Biologie, 63:11-6.
Xu XM, Yu JP, He XF, Li JH, Yu LL, Yu HG .2005. Effects of garlicin on apoptosis in rat model of colitis. WJG, 11:4579-4582.
Yu Y-M, Chao T-Y, Chang W-C, Chang MJ, Lee M-F.2016. Thymol reduces oxidative stress, aortic intimal thickening, and inflammation-related gene expression in hyperlipidemic rabbits. J Food Drug Anal, 24:556-563.