Abstract
Objectives
Ulcerative colitis (UC) is a non-specific intestinal inflammatory disease. Several studies demonstrated that inflammation and oxidative stress play significant role in the pathogenesis of this disease. This study aimed to determine the protective effect and possible mechanism by which stevia affects the course of experimentally induced colitis.
Methods
Male rats were received stevia 20, 40, 80 mg/kg/day before induction of colitis by intra-rectal administration of 2 mL of 4% acetic acid, AA. Macroscopic and histopathological examination of the colon were done. Colonic content of catalase (CAT), superoxide dismutase (SOD), reduced glutathione (GSH), myeloperoxidase (MPO) and thiobarbituric acid reactive substances (TBARS) activities and serum levels of interleukin (IL)1- β and tumor necrosis factor (TNF)-α were assessed. Real time-PCR (RT-PCR) was done to determine the expression of NF-κB, Nrf2 and PPARγ genes. Spontaneous contraction and effects of increasing concentrations of acetylcholine and stevia have been studied on the isolated colonic segments.
Results
Stevia ameliorated colitis not only histopathologically but also it decreased the level of TNF-α, IL-1β, TBARS, MPO and the expression of NF-κB which were significantly increased in the AA group. The concentration of GSH, SOD, CAT and expression of Nrf2 and PPARγ were significantly increased with stevia. Moreover, stevia showed a relaxant effect on the colonic contractility which was increased in AA group. These all effects of stevia were more prominent with its highest dose.
Conclusion
Our results explored that, stevia acts protectively against UC by its anti-inflammatory and antioxidant properties which mediated by up-regulation of Nrf2 and PPARγ with downregulation of NF-κB. We suggest that stevia has the potential for treatment of chronic inflammatory diseases, such as UC.
Acknowledgments
The Physiology Department, Faculty of Medicine, Mansoura University and Department of Forensic Medicine, and Toxicology, Faculty of Veterinary Medicine, Mansoura University acknowledged for their contribution in the experimental part of the present study.
-
Research funding: There was no fund to support the current work .
-
Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.
-
Competing interests: Authors state no conflict of interest.
-
Ethical approval: Our Local Committee of Animal Care and Used approved this protocol (Code number R.19.11.663).
References
1. Liu, Y, Wang, X, Hu, CAA. Therapeutic potential of amino acids in inflammatory bowel disease. Nutrients 2017;9:1–18. https://doi.org/10.3390/nu9090920.Search in Google Scholar
2. Reiff, C, Kelly, D. Inflammatory bowel disease, guts bacteria and probiotic therapy. Int J Med Microbiol 2010;300:25–33. https://doi.org/10.1016/j.ijmm.2009.08.004.Search in Google Scholar
3. Seidelin, JB. Regulation of antiapoptotic and cytoprotective pathways in colonic epithelial cells in ulcerative colitis. Scand J Gastroenterol 2015;50(1 Suppl):1–29. https://doi.org/10.3109/00365521.2016.1101245.Search in Google Scholar
4. Cetinkaya, A, Bulbuloglu, E, Kantarceken, B, Ciralik, H, Kurutas, EB, Buyukbese, MA, et al. Effects of L-carnitine on oxidant/antioxidant status in acetic acid-induced colitis. Dig Dis Sci 2006;51:488–94. https://doi.org/10.1007/s10620-006-3160-9.Search in Google Scholar
5. Qiu, W, Wu, B, Wang, X, Buchanan, ME, Regueiro, MD, Hartman, DJ, et al. PUMA-mediated intestinal epithelial apoptosis contributes to ulcerative colitis in humans and mice. J Clin Invest 2011;121:1722–32. https://doi.org/10.1172/jci42917.Search in Google Scholar
6. Lemus-Mondaca, R, Vega-Galvez, A, Zura-Bravo, L, Ah-Hen, K. Stevia rebaudiana Bertoni, source of a high-potency natural sweetener: a comprehensive review on the biochemical, nutritional and functional aspects. Food Chem 2012;132:1121–32. https://doi.org/10.1016/j.foodchem.2011.11.140.Search in Google Scholar
7. Shiozaki, K, Nakano, T, Yamaguchi, T, Sato, M, Sato, N. The protective effect of stevia extract on the gastric mucosa of rainbow trout Oncorhynchus mykiss (Walbaum) fed dietary histamine. Aquacult Res 2004;35:1421–8. https://doi.org/10.1111/j.1365-2109.2004.01164.x.Search in Google Scholar
8. Zhu, Y, Alvares, K, Huang, Q, Rao, MS, Reddy, JK. Cloning of a new member of the peroxisome proliferator-activated receptor gene family from mouse liver. J Biol Chem 1993;268:26817–20.10.1016/S0021-9258(19)74184-2Search in Google Scholar
9. Matera, S, Piersante, MV, Ragone, MI, Consolini, AE. Sedative and antispasmodic effects of Stevia rebaudiana and noncompetitive inhibition of intestinal contractility by stevioside. Pharmacologyonline 2012;1:1–8.Search in Google Scholar
10. Lefebvre, AM, Chen, I, Desreumaux, P, Najib, J, Fruchart, JC, Geboes, K, et al. Activation of the peroxisome proliferator-activated receptor gamma promotes the development of colon tumors in C57BL/6J-APCMin/+ mice. Nat Med 1998;4:1053–7. https://doi.org/10.1038/2036.Search in Google Scholar PubMed
11. Lu, MC, Ji, JA, Jiang, YL, Chen, ZY, Yuan, ZW, You, QD, et al. An inhibitor of the Keap1-Nrf2 protein-protein interaction protects NCM460 colonic cells and alleviates experimental colitis. Sci Rep [Internet] 2016;6:1–13. Available from: https://doi.org/10.1038/srep26585.Search in Google Scholar PubMed PubMed Central
12. Aleksunes, LM, Manautou, JE. Emerging role of Nrf2 in protecting against hepatic and gastrointestinal disease. Toxicol Pathol 2007;35:459–73. https://doi.org/10.1080/01926230701311344.Search in Google Scholar
13. Copple, IM, Goldring, CE, Kitteringham, NR, Park, BK. The Nrf2-Keap1 defence pathway: role in protection against drug-induced toxicity. Toxicology 2008;246:24–33. https://doi.org/10.1016/j.tox.2007.10.029.Search in Google Scholar
14. Khor, TO, Huang, M-T, Kwon, KH, Chan, JY, Reddy, BS, Kong, A-N. Nrf2-deficient mice have an increased susceptibility to dextran sulfate sodium-induced colitis. Canc Res 2006;66:11580–4. https://doi.org/10.1158/0008-5472.can-06-3562.Search in Google Scholar
15. Huang, J, Tabbi-Anneni, I, Gunda, V, Wang, L. Transcription factor Nrf2 regulates SHP and lipogenic gene expression in hepatic lipid metabolism. Am J Physiol Gastrointest Liver Physiol 2010;299:G1211–21. https://doi.org/10.1152/ajpgi.00322.2010.Search in Google Scholar
16. Cho, H-Y, Gladwell, W, Wang, X, Chorley, B, Bell, D, Reddy, SP, et al. Nrf2-regulated PPAR{gamma} expression is critical to protection against acute lung injury in mice. Am J Respir Crit Care Med 2010;182:170–82. https://doi.org/10.1164/rccm.200907-1047oc.Search in Google Scholar
17. Chorley, BN, Campbell, MR, Wang, X, Karaca, M, Sambandan, D, Bangura, F, et al. Identification of novel NRF2-regulated genes by ChIP-Seq: influence on retinoid X receptor alpha. Nucleic Acids Res 2012;40:7416–29. https://doi.org/10.1093/nar/gks409.Search in Google Scholar
18. Lee, C. Collaborative power of Nrf2 and PPARgamma activators against metabolic and drug-induced oxidative injury. Oxid Med Cell Longev 2017;2017:1378175. https://doi.org/10.1155/2017/1378175.Search in Google Scholar
19. Millar, AD, Rampton, DS, Chander, CL, Claxson, AW, Blades, S, Coumbe, A, et al. Evaluating the antioxidant potential of new treatments for inflammatory bowel disease using a rat model of colitis. Gut 1996;39:407–15. https://doi.org/10.1136/gut.39.3.407.Search in Google Scholar
20. Misra, H, Soni, M, Silawat, N, Mehta, D, Mehta, BK, Jain, DC. Antidiabetic activity of medium-polar extract from the leaves of Stevia rebaudiana Bert. (Bertoni) on alloxan-induced diabetic rats. J Pharm Bio Allied Sci 2011;3:242–8. https://doi.org/10.4103/0975-7406.80779.Search in Google Scholar
21. Sinha, AK. Colorimetric assay of catalase. Anal Biochem 1972;47:389–94. https://doi.org/10.1016/0003-2697(72)90132-7.Search in Google Scholar
22. Kakkar, P, Das, B, Viswanathan, PN. A modified spectrophotometric assay of superoxide dismutase. Indian J Biochem Biophys 1984;21:130–2. PMID: 6490072.Search in Google Scholar
23. Ellman, GL. Tissue sulfhydryl groups. Arch Biochem Biophys 1959;82:70–7. https://doi.org/10.1016/0003-9861(59)90090-6.Search in Google Scholar
24. Krawisz, JE, Sharon, P, Stenson, WF. Quantitative assay for acute intestinal inflammation based on myeloperoxidase activity. Assessment of inflammation in rat and hamster models. Gastroenterology 1984;87:1344–50. https://doi.org/10.1016/0016-5085(84)90202-6.Search in Google Scholar
25. Parlakpinar, H, Ozer, MK, Sahna, E, Vardi, N, Cigremis, Y, Acet, A. Amikacin-induced acute renal injury in rats: protective role of melatonin. J Pineal Res 2003;35:85–90. https://doi.org/10.1034/j.1600-079x.2003.00059.x.Search in Google Scholar
26. Kim, SJ, Park, JH, Song, DK, Park, KS, Lee, JE, Kim, ES, et al. Alterations of colonic contractility in long-term diabetic rat model. J Neurogastroenterol Motil 2011;17:372–80. https://doi.org/10.5056/jnm.2011.17.4.372.Search in Google Scholar
27. Vergara-Galicia, J, Aguirre-Crespo, F, Castillo-Espana, P, Arroyo-Mora, A, Lopez-Escamilla, AL, Villalobos-Molina, R, et al. Micropropagation and vasorelaxant activity of Laelia autumnalis (Orchidaceae). Nat Prod Res 2010;24:106–14. https://doi.org/10.1080/14786410802340820.Search in Google Scholar
28. Gaudio, E, Taddei, G, Vetuschi, A, Sferra, R, Frieri, G, Ricciardi, G, et al. Dextran sulfate sodium (DSS) colitis in rats: clinical, structural, and ultrastructural aspects. Dig Dis Sci 1999;44:1458–75. https://doi.org/10.1023/a:1026620322859.10.1023/A:1026620322859Search in Google Scholar
29. D’Argenio, G, Mazzone, G, Tuccillo, C, Ribecco, MT, Graziani, G, Gravina, AG, et al. Apple polyphenols extract (APE) improves colon damage in a rat model of colitis. Dig Liver Dis 2012;44:555–62. https://doi.org/10.1016/j.dld.2012.01.009.Search in Google Scholar
30. El-Abhar, HS, Hammad, LNA, Gawad, HSA. Modulating effect of ginger extract on rats with ulcerative colitis. J Ethnopharmacol 2008;118:367–72. https://doi.org/10.1016/j.jep.2008.04.026.Search in Google Scholar
31. Harputluoglu, MM, Demirel, U, Yucel, N, Karadag, N, Temel, I, Firat, S, et al. The effects of Gingko biloba extract on acetic acid-induced colitis in rats. Turk J Gastroenterol 2006;17:177–82. PMID: 16941250.Search in Google Scholar
32. Pavlick, KP, Laroux, FS, Fuseler, J, Wolf, RE, Gray, L, Hoffman, J, et al. Role of reactive metabolites of oxygen and nitrogen in inflammatory bowel disease. Free Radic Biol Med 2002;33:311–22. https://doi.org/10.1016/s0891-5849(02)00853-5.Search in Google Scholar
33. Sakthivel, KM, Guruvayoorappan, C. Modulating effect of Biophytum sensitivum extract on rats with acetic acid-induced ulcerative colitis. Pharm Biol 2014;52:1570–80. https://doi.org/10.3109/13880209.2014.908396.Search in Google Scholar PubMed
34. Tahan, G, Aytac, E, Aytekin, H, Gunduz, F, Dogusoy, G, Aydin, S, et al. Vitamin E has a dual effect of anti-inflammatory and antioxidant activities in acetic acid-induced ulcerative colitis in rats. Can J Surg 2011;54:333–8. https://doi.org/10.1503/cjs.013610.Search in Google Scholar PubMed PubMed Central
35. Stucchi, A, Reed, K, O’Brien, M, Cerda, S, Andrews, C, Gower, A, et al. A new transcription factor that regulates TNF-alpha gene expression, LITAF, is increased in intestinal tissues from patients with CD and UC. Inflamm Bowel Dis 2006;12:581–7. https://doi.org/10.1097/01.mib.0000225338.14356.d5.Search in Google Scholar PubMed
36. Boonkaewwan, C, Burodom, A. Anti-inflammatory and immunomodulatory activities of stevioside and steviol on colonic epithelial cells. J Sci Food Agric 2013;93:3820–5. https://doi.org/10.1002/jsfa.6287.Search in Google Scholar PubMed
37. Kruidenier, L, Verspaget, HW. Review article: oxidative stress as a pathogenic factor in inflammatory bowel disease-radicals or ridiculous? Aliment Pharmacol Ther 2002;16:1997–2015. https://doi.org/10.1046/j.1365-2036.2002.01378.x.Search in Google Scholar PubMed
38. Hartmann, RM, Morgan Martins, MI, Tieppo, J, Fillmann, HS, Marroni, NP. Effect of Boswellia serrata on antioxidant status in an experimental model of colitis rats induced by acetic acid. Dig Dis Sci 2012;57:2038–44. https://doi.org/10.1007/s10620-012-2134-3.Search in Google Scholar PubMed
39. Isozaki, Y, Yoshida, N, Kuroda, M, Takagi, T, Handa, O, Kokura, S, et al. Effect of a novel water-soluble vitamin E derivative as a cure for TNBS-induced colitis in rats. Int J Mol Med 2006;17:497–502. https://doi.org/10.3892/ijmm.17.3.497.Search in Google Scholar
40. Aleisa, AM, Al-Rejaie, SS, Abuohashish, HM, Ola, MS, Parmar, MY, Ahmed, MM. Pretreatment of Gymnema sylvestre revealed the protection against acetic acid-induced ulcerative colitis in rats. BMC Compl Alternative Med 2014;14:49. https://doi.org/10.1186/1472-6882-14-49.Search in Google Scholar PubMed PubMed Central
41. Tanida, S, Mizoshita, T, Mizushima, T, Sasaki, M, Shimura, T, Kamiya, T, et al. Involvement of oxidative stress and mucosal addressin cell adhesion molecule-1 (MAdCAM-1) in inflammatory bowel disease. J Clin Biochem Nutr 2011;48:112–6. https://doi.org/10.3164/jcbn.10-41.Search in Google Scholar PubMed PubMed Central
42. Choudhary, S, Keshavarzian, A, Yong, S, Wade, M, Bocckino, S, Day, BJ, et al. Novel antioxidants zolimid and AEOL11201 ameliorate colitis in rats. Dig Dis Sci 2001;46:2222–30. https://doi.org/10.1023/a:1011975218006.10.1023/A:1011975218006Search in Google Scholar
43. Lu, P-D, Zhao, Y-H. Targeting NF-kappaB pathway for treating ulcerative colitis: comprehensive regulatory characteristics of Chinese medicines. Chin Med 2020;15:15. https://doi.org/10.1186/s13020-020-0296-z.Search in Google Scholar PubMed PubMed Central
44. Hagar, HH, El-Medany, A, El-Eter, E, Arafa, M. Ameliorative effect of pyrrolidinedithiocarbamate on acetic acid-induced colitis in rats. Eur J Pharmacol 2007;554:69–77. https://doi.org/10.1016/j.ejphar.2006.09.066.Search in Google Scholar PubMed
45. Wang, A, Keita, AV, Phan, V, McKay, CM, Schoultz, I, Lee, J, et al. Targeting mitochondria-derived reactive oxygen species to reduce epithelial barrier dysfunction and colitis. Am J Pathol 2014;184:2516–27. https://doi.org/10.1016/j.ajpath.2014.05.019.Search in Google Scholar PubMed PubMed Central
46. Sun, JM, Wang, CM, Guo, Z, Hao, YY, Xie, YJ, Gu, J, et al. Reduction of isoproterenol-induced cardiac hypertrophy and modulation of myocardial connexin43 by a KATPchannel agonist. Mol Med Rep 2015;11:1845–50. https://doi.org/10.3892/mmr.2014.2988.Search in Google Scholar PubMed
47. Yang, Y, Cai, X, Yang, J, Sun, X, Hu, C, Yan, Z, et al. Chemoprevention of dietary digitoflavone on colitis-associated colon tumorigenesis through inducing Nrf2 signaling pathway and inhibition of inflammation. Mol Canc 2014;13:48. https://doi.org/10.1186/1476-4598-13-48.Search in Google Scholar PubMed PubMed Central
48. Wen, Z, Liu, W, Li, X, Chen, W, Liu, Z, Wen, J, et al. A protective role of the NRF2-Keap1 pathway in maintaining intestinal barrier function. Oxid Med Cell Longev 2019;2019:1759149. https://doi.org/10.1155/2019/1759149.Search in Google Scholar PubMed PubMed Central
49. Pandurangan, AK, Mohebali, N, Norhaizan, ME, Looi, CY. Gallic acid attenuates dextran sulfate sodium-induced experimental colitis in BALB/c mice. Drug Des Dev Ther 2015;9:3923–34. https://doi.org/10.2147/dddt.s86345.Search in Google Scholar
50. Cuadrado, A, Manda, G, Hassan, A, Alcaraz, MJ, Barbas, C, Daiber, A, et al. Transcription factor NRF2 as a therapeutic target for chronic diseases: a systems medicine approach. Pharmacol Rev 2018;70:348–83. https://doi.org/10.1124/pr.117.014753.Search in Google Scholar PubMed
51. Lu, M-C, Ji, J-A, Jiang, Y-L, Chen, Z-Y, Yuan, Z-W, You, Q-D, et al. An inhibitor of the Keap1-Nrf2 protein-protein interaction protects NCM460 colonic cells and alleviates experimental colitis. Sci Rep 2016;6:26585. https://doi.org/10.1038/srep26585.Search in Google Scholar PubMed PubMed Central
52. Hontecillas, R, Bassaganya-Riera, J. Peroxisome proliferator-activated receptor gamma is required for regulatory CD4+ T cell-mediated protection against colitis. J Immunol 2007;178:2940–9. https://doi.org/10.4049/jimmunol.178.5.2940.Search in Google Scholar PubMed
53. Jiang, J, Qi, L, Lv, Z, Jin, S, Wei, X, Shi, F. Dietary stevioside supplementation alleviates lipopolysaccharide-induced intestinal mucosal damage through anti-inflammatory and antioxidant effects in broiler chickens. Antioxidants 2019;8:575. https://doi.org/10.3390/antiox8120575.Search in Google Scholar PubMed PubMed Central
54. Ramos-Tovar, E, Flores-Beltrán, RE, Galindo-Gómez, S, Camacho, J, Tsutsumi, V, Muriel, P. An aqueous extract of stevia rebaudiana variety Morita II prevents liver damage in a rat model of cirrhosis that mimics the human disease. Ann Hepatol 2019;18:472–9. https://doi.org/10.1016/j.aohep.2018.10.002.Search in Google Scholar PubMed
55. Ramos-Tovar, E, Hernández-Aquino, E, Casas-Grajales, S, Buendia-Montaño, LD, Galindo-Gómez, S, Camacho, J, et al. Stevia prevents acute and chronic liver injury induced by carbon tetrachloride by blocking oxidative stress through Nrf2 upregulation. Oxid Med Cell Longev 2018;2018:1–12 https://doi.org/10.1155/2018/3823426.Search in Google Scholar PubMed PubMed Central
56. Schwab, M, Reynders, V, Loitsch, S, Shastri, YM, Steinhilber, D, Schroder, O, et al. PPARgamma is involved in mesalazine-mediated induction of apoptosis and inhibition of cell growth in colon cancer cells. Carcinogenesis 2008;29:1407–14. https://doi.org/10.1093/carcin/bgn118.Search in Google Scholar PubMed
57. Yamamoto-Furusho, JK, Peñaloza-Coronel, A, Sánchez-Muñoz, F, Barreto-Zuñiga, R, Dominguez-Lopez, A. Peroxisome proliferator-activated receptor-gamma (PPAR-γ) expression is down regulated in patients with active ulcerative colitis. England: Inflammatory bowel diseases; 2011, vol 17:680–1.10.1002/ibd.21322Search in Google Scholar PubMed
58. Saubermann, LJ, Nakajima, A, Wada, K, Zhao, S, Terauchi, Y, Kadowaki, T, et al. Peroxisome proliferator-activated receptor gamma agonist ligands stimulate a Th2 cytokine response and prevent acute colitis. Inflamm Bowel Dis 2002;8:330–9. https://doi.org/10.1097/00054725-200209000-00004.Search in Google Scholar PubMed
59. Bassotti, G, Antonelli, E, Villanacci, V, Baldoni, M, Dore, MP. Colonic motility in ulcerative colitis. United Eur Gastroenterol J 2014;2:457–62. https://doi.org/10.1177/2050640614548096.Search in Google Scholar PubMed PubMed Central
60. Hosseini, JM, Goldhill, JM, Bossone, C, Pineiro-Carrero, V, Shea-Donohue, T. Progressive alterations in circular smooth muscle contractility in TNBS-induced colitis in rats. Neuro Gastroenterol Motil 1999;11:347–56. https://doi.org/10.1046/j.1365-2982.1999.00165.x.Search in Google Scholar PubMed
61. Goldhill, JM, Sanders, KM, Sjogren, R, Shea-Donohue, T. Changes in enteric neural regulation of smooth muscle in a rabbit model of small intestinal inflammation. Am J Physiol 1995;268:G823–30. https://doi.org/10.1152/ajpgi.1995.268.5.g823.Search in Google Scholar
62. Bossone, C, Hosseini, JM, Pineiro-Carrero, V, Shea-Donohue, T. Alterations in spontaneous contractions in vitro after repeated inflammation of rat distal colon. Am J Physiol Gastrointest Liver Physiol 2001;280:G949–57. https://doi.org/10.1152/ajpgi.2001.280.5.g949.Search in Google Scholar
63. Shiozaki, K, Fujii, A, Nakano, T, Yamaguchi, T, Sato, M. Inhibitory effects of hot water extract of the stevia stem on the contractile response of the smooth muscle of the guinea pig ileum. Biosci Biotechnol Biochem 2006;70:489–94. https://doi.org/10.1271/bbb.70.489.Search in Google Scholar PubMed
64. Lee, CN, Wong, KL, Liu, JC, Chen, YJ, Cheng, JT, Chan, P. Inhibitory effect of stevioside on calcium influx to produce antihypertension. Planta Med 2001;67:796–9. https://doi.org/10.1055/s-2001-18841.Search in Google Scholar PubMed
© 2020 Walter de Gruyter GmbH, Berlin/Boston
