Protective effect of royal jelly in 2,4,6 trinitrobenzene sulfonic acid-induced colitis in rats

Document Type : Original Article


1 University of Trakya, Faculty of Medicine, Department of Histology- Embryology, Balkan Campus, 22030, Edirne, Turkey

2 University of Trakya, Faculty of Medicine, Department of Gastroenterolgy, Balkan Campus, 22030, Edirne, Turkey


Objective(s):In the present study, we evaluated immunological and immunomodulatory properties of royal jelly (RJ) in 2,4,6 trinitrobenzene sulfonic acid (TNBS)-induced colitis in rats.
Materials and Methods: Eighteen adult female Wistar albino rats were divided into three groups of six animals each: a control group that received only saline solution, a TNBS-induced colitis group, and a TNBS-colitis+RJ group that received 250 mg/kg/day of RJ for seven days before the induction of colitis, following by the same treatment for an additional seven days. At the end of the experiment, cardiac blood and colon samples were obtained under deep anaesthesia from the animals in all groups. Serum interleukin-1β (IL-1β), tumour necrosis factor-alpha (TNF-α) and IL-10 levels were analyzed with an enzyme-linked immunosorbent assay (ELISA). Five-micrometre-thick sections were stained with haematoxylin-eosin (H&E) for microscopic evaluations. For immunohistochemical evaluations, the paraffin sections were stained with anti-CD3 (cluster of differentiation), anti-CD5, anti-CD8 and anti-CD45.
Results: The results showed that the oral RJ treatment inhibited proinflammatory cytokines, IL-1β and TNF-α secretion, while increasing anti-inflammatory cytokine IL-10 production in the TNBS-induced colitis+RJ group compared with the colitis group not treated with RJ. The colitis was not as severe in the colitis+RJ group, with ulcerative damage, weight loss and inflammatory scores suggesting that impaired CD3-, CD5-, CD8- and CD45-positive T cell immune responses likely mediated the anti-inflammatory effect.
Conclusion: The antioxidant and anti-inflammatory properties of RJ protected colon mucosa against TNBS-induced colitis in rats orally treated with RJ.


1.Abboud PA, Hake PW, Burroughs TJ, Odoms K, O'Connor M, Mangeshkar P, et al. Therapeutic effect of epigallocatechin-3-gallate in a mouse model of colitis. Eur J Pharmacol 2008; 579:411–417.
2.Safarpour AR, Hosseini SV, Mehrabani D. Epidemiology of inflammatory bowel diseases in Iran and Asia; A mini review. Iran J Med Sci 2013; 38:140-149.  
3.Carvalho AT, Souza H, Carneiro AJ, Castelo-Branco M, Madi K, Schanaider A, et al. Therapeutic and prophy-lactic thalidomide in TNBS-induced colitis: Synergistic effects on TNF-α, IL-12 and VEGF production. World J Gastroenterol 2007; 13:2166-2173.
4. Ko JK, Auyeung KK. Inflammatory bowel disease: etiology, pathogenesis and current therapy. Curr Pharm Des 2014; 20(7):1082-1096.
5.Mazzon E, Muia C, Di Paola R, Genovese T, De Sarro A, Cuzzocrea S. Thalidomide Treatment reduces colon injury induced by experimental colitis. Shock 2005; 23(6):556–564.
6.Prakash A, Medhi B, Avti PK, Saikia UN, Pandhi P, Khanduja KL. Effect of different doses of manuka honey in experimentally induced inflammatory bowel disease in rats. Phytother Res 2008; 22:1511–1519.
7.Münstedt K, Bargello M, Hauenschild A. Royal jelly reduces the serum glucose levels in healthy subjects. J Med Food 2009; 12(5):1170-1172.
8.Viuda-Martos M, Ruiz-Navajas Y, Fernandez-Lopez J, Perez-Alvarez JA. Functional properties of honey, propolis, and royal jelly. J Food Sci 2008; 73(9):R117–R124.
9.Kohno K, Okamoto I, Sano O, Arai N, Iwaki K, Ikeda M, et al. Royal jelly inhibits the production of proinflammatory cytokines by activated macrophages. Biosci Biotechnol Biochem 2004; 68:138–145.
10.Karaca T, Şimşek N, Uslu S, Kalkan Y, Can I, Kara A, et al. The effect of royal jelly on CD3(+), CD5(+), CD45(+) T-cell and CD68(+) cell distribution in the colon of rats with acetic acid-induced colitis. Allergol Immunopathol (Madr) 2012; 40:357-361.
11.Gasic S, Vucevic D, Vasilijic S, Antunovic M, Chinou I, Colic M. Evaluation of the immunomodula-tory activities of royal jelly components in vitro. Immunopharmacol Immunotoxicol 2007; 29:521–536.
12.Karaca T, Bayiroglu F, Yoruk M, Kaya MS, Uslu S, Comba B, et al. Effect of royal jelly on experimental colitis induced by acetic acid and alteration of mast cell distribution in the colon of rats. Eur J Histochem 2010; 54(4): e.35.
13.Mehrabani D, Bahrami F, Hosseini SV, Ashraf MJ, Tanideh N, Rezaianzadeh A, et al. The healing effect of Teucrium polium in acetic acid induced ulcerative colitis in the dog as an animal model. Middle East J Dig Dis 2012;4:41-48.
14.Tanideh N, Akbari Baseri F, Jamshidzadeh A, Ashraf MJ, Kuhi O, Mehrabani D. The healing effect of strawberry extract on acetic acid-induced ulcerative colitis in rat. World Appl Sci J 2014; 31(3):281-288.
15.Mazzon E, Muià C, Paola RD, Genovese T, Menegazzi M, De Sarro A et al. Green tea polyphenol extract attenuates colon injury induced by experimental colitis. Free Radic Res 2005; 39(9):1017–1025.
16.McCafferty DM, Sharkey KA, Wallace JL. Beneficial effects of local or systemic lidocaine in experimental colitis. Am J Physiol - Gas Liv Physiol 1994; 266:G560-G567.
17.Obermeier F, Kojouharoff G, Hans W, Schölmerich J, Gross V, Falk W. Interferon-gamma (IFN-γ)- and tumour necrosis factor (TNF)-induced nitric oxide as toxic effector molecule in chronic dextran sulphate sodium (DSS)-induced colitis in mice. Clin Exp Immunol 1999; 116:238–245.
18.Keshavarz A,  Minaiyan M, Ghannadi A, Mahzouni P. Effects of Carum carvi L. (Caraway) extract and essential oil on TNBS-induced colitis in rats. Res Pharm Sci 2013; 8:1-8.
19.Socca EA, Luiz-Ferreira A, de Faria FM, de Almeida AC, Dunder RJ, Manzo LP, Brito AR. Inhibition of tumor necrosis factor-alpha and cyclooxigenase-2 by Isatin: A molecular mechanism of protection against TNBS-induced colitis in rats. Chem Biol Interact 2014; 209:48–55.
20.Paiotti AP, Miszputen SJ, Oshima CT, de Oliveira Costa H, Ribeiro DA, Franco M. Effect of COX-2 inhibitor after TNBS-induced colitis in Wistar rats. J Mol Histol 2009; 40:317–324.
21.Bilsel Y, Bugra D, Yamaner S, Bulut Y, Cevikbas U, Turkoglu U. Could honey have a place in colitis therapy? Effects of honey, prednisolone, and disulfiram on inflammation, nitric oxide, and free radical formation. Dig Surg 2002; 19:306–311.
22.Oka H, Emori Y, Kobayashi N, Hayashi Y, Nomoto K. Suppression of allergic reactions by royal jelly in association with the restoration of macrophage function and the improvement of Th1/Th2 cell responses. Int Immunopharmacol 2001; 1:521-532.
23.Mehrabani D, Ziaei M, Hosseini SV, Ghahramani L, Bananzadeh AM, Ashraf MJ, et al. The effect of calendula officinalis in therapy of acetic acid induced ulcerative colitis in dog as an animal model. Iran Red Crescent Med J 2011; 13(12):884-890.
24.Shen W, Durum S. Synergy of IL-23 and Th17 cytokines: New light on inflammatory bowel disease.  Neurochem Res 2010; 35:940–946.
25.Elson CO, Cong Y, Weaver CT, Schoeb TR, McClanahan TK, Fick RB, et al. Monoclonal anti-interleukin 23 reverses active colitis in a T cell-mediated model in mice. Gastroenterology 2007; 132:2359–2370.
26.Fazio L, Cavazza E, Spisni E, Strillacci A, Centanni M, Candela M, et al. Longitudinal analysis of inflam-mation and microbiota dynamics in a model of mild chronic dextran sulfate sodium-induced colitis in mice. World J Gastroenterol 2014; 20:2051-2061.
27.Celinski K, Dworzanski T, Fornal R, Korolczuk A, Madro A, Slomka M. Comparison of the anti-inflammatory and therapeutic actions of ppar-Gammaagonists rosiglitazone and troglitazone in experimental colitis. J Physiol Pharmacol 2012; 63:631-640.
28.Celinski K, Dworzanski T, Fornal R, Korolczuk A, Madro A, Slomka M. Comparsion of anti-nflammatory properties of peroxisome proliferator-activated recep-tor gammaagonists rosiglitazone and troglitazone in prophylactic treatment of experimental colitis. J Physiol Pharmacol 2013; 64:587-595.
29.Nagai T, Inoue R, Suzuki N, Nagashima T. Anti-oxidant properties of enzymatic hydrolysates from royal jelly. J Med Food 2006; 9:363-367.
30.Peng X, Li X, Wang W, Li N, Ma J, Shen S. Expre-ssion of Th1/Th2 inflammatory cytokines in rat treatment model of ulcerative colitis (Abstract). J Central South Univ 2013; 38:1020-1028.
31.Wang K, Xuan X, Wang L, Tong L, Huang Q, Zhu L, et al. Expression and correlation analysis between inflammatory cytokines and calprotectin in the rat model of ulcerative colitis. Chinese J Cell Mol Immunol 2014; 30:278-280.
32.Fujii A, Kobayashi S, Kuboyama N, Furukawa Y, Kaneko Y, Ishihama S, et al. Augmentation of wound healing by royal jelly (RJ) in streptozotocin-diabetic rats. Jpn J Pharmacol 1990; 53:331–337.
33.Okamoto I, Taniguchi Y, Kunikata T, Kohno K, Iwaki K, Ikeda M, et al. Major royal jelly protein 3 modulates immune responses in vitro and in vivo. Life Sci J 2003; 73:2029-2045.
34.Vucevic D, Melliou E, Vasilijic S, Gasic S, Ivanovski P, Chinou I, et al. Fatty acids isolated from royal jelly modulate dendritic cell-mediated immune response in vitro. Int Immunopharmacol 2007; 7:1211–1220.
35.Fiorani M, Accorsi A, Blasa M, Diamantini G, Piatti E. Flavonoids from Italian multifloral honeys reduce the extracellular ferricyanide in human red blood cells. J Agric Food Chem 2006; 54:8328-8334.
36.Mehrabani D, Almasi-Hashiani A, Moshfeghi K, Khedmati E. Survival rate and its predictors in colorectal cancer patients, Southern Iran. Middle East J Sci Res 2012; 12:1072-1077.