Journal List > J Nutr Health > v.52(2) > 1122025

J Nutr Health. 2019 Apr;52(2):139-148. Korean.
Published online Apr 30, 2019.  https://doi.org/10.4163/jnh.2019.52.2.139
© 2019 The Korean Nutrition Society
Anti-inflammatory effects of mulberry twig extracts on dextran sulfate sodium-induced colitis mouse model
Xuelei Cui and Eunjung Kim
Department of Food Science and Nutrition, Daegu Catholic University, Gyeongsan, Gyeongbuk 38430, Korea.

To whom correspondence should be addressed. tel: +82-53-850-3523, Email: kimeunj@cu.ac.kr
Received Mar 01, 2019; Revised Mar 15, 2019; Accepted Mar 18, 2019.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.


Abstract

Purpose

Ulcerative colitis is a common inflammatory bowel disease. Prolonged colitis can be a risk factor for the development of colorectal cancer. Mulberry twig (MT, Sangzhi), a dry branch of Morus alba L., which is widely distributed throughout East Asia, has been shown to have anti-inflammatory activities in the cells. However, the effects of MT extracts on colitis in in vivo are limited. Therefore, in this study, we investigated the anti-inflammatory effects of MT extracts in the dextran sulfate sodium (DSS)-induced mouse colitis model.

Methods

Six week-old, male ICR mice were divided into 3 groups: Control (n = 5), DSS (n = 7), and DSS+MT (n = 7) groups. Mice in the DSS and DSS+MT groups were administrated 3% DSS in drinking water for 5 days to induce colitis. At the same time, water extracts of MT (5 g/kg body weight/day) were orally administered to mice in the DSS+MT groups for 5 days.

Results

The MT extracts significantly reduced the clinical and pathological characteristics of colitis. Disease activity index, mucosal thickness, and colonocyte proliferation were significantly reduced in the DSS+MT group compared with the DSS group. Furthermore, MT administration reduced the levels of plasma TNF-α, IL-6, and the colonic myeloperoxidase activity as well as mRNA expression of TNF-α, IL-6, Cox-2, and iNOS.

Conclusion

Taken together, these results suggest that MT water extracts have potent anti-colitis activities in the mouse colitis model.

Keywords: mulberry twig; Morus alba L.; colitis; dextran sulfate sodium; mouse

Figures


Fig. 1
Effects of MT on disease activity index, colon length and colon weight. (A) Disease activity index was measured daily during DSS administration. Large intestine was obtained after the DSS administration and their lengths (B) and weight (C) were measured. Value are presented as the mean ± SE. Means with different letters are significantly different at p < 0.05 by Duncan's multiple range test. Control, group received water without DSS; DSS, group received 3% DSS in drinking water for 5 days; DSS+MT, group received 3% DSS and oral administration with MT water extracts (5 g/kg/day) for 5 days; DSS, dextran sulfate sodium; MT, mulberry twig; NS, not significant
Click for larger image


Fig. 2
Histopathological change of large intestine. The rectum of mouse was stained with hematoxylin and eosin. (A) Representative histological images of Control, DSS, and DSS+MT groups are shown. (B) Mucosal thickness was measured microscopically. Value are presented as the mean ± SE. Means with different letters are significantly different at p < 0.05 by Duncan's multiple range test. Control, group received water without DSS; DSS, group received 3% DSS in drinking water for 5 days; DSS+MT, group received 3% DSS and oral administration with MT water extracts (5 g/kg/day) for 5 days; DSS, dextran sulfate sodium; MT, mulberry twig
Click for larger image


Fig. 3
Proliferation rate of colonocytes. BrdU incorporation was demonstrated immunohistochemically after i.p. injection of BrdU. The number of BrdU-labeled cells were counted and labeling index of BrdU was determined (BrdU-labeled cells/300 cells). Value are presented as the mean ± SE. Means with different letters are significantly different at p < 0.05 by Duncan's multiple range test. Control, group received water without DSS; DSS, group received 3% DSS in drinking water for 5 days; DSS+MT, group received 3% DSS and oral administration with MT water extracts (5 g/kg/day) for 5 days; DSS, dextran sulfate sodium; MT, mulberry twig
Click for larger image


Fig. 4
The levels of inflammatory mediators in colorectal tissue and in plasma. (A) TNF-α and IL-6 mRNA expression in colorectal region were measured by qRT-PCR. (B) The levels of plasma NO, TNF-α and IL-6 were determined in experimental groups. Value are presented as the mean ± SE. Means with different letters are significantly different at p < 0.05 by Duncan's multiple range test. Control, group received water without DSS; DSS, group received 3% DSS in drinking water for 5 days; DSS+MT, group received 3% DSS and oral administration with MT water extracts (5 g/kg/day) for 5 days; DSS, dextran sulfate sodium; MT, mulberry twig
Click for larger image


Fig. 5
Colorectal inflammatory gene expression and MPO activity. (A) Cox-2 and iNOS mRNA expression in the distal part of large intestine was analyzed by qRT-PCR. (B) Total protein lysates from the distal part of large intestine were subjected to western analysis with their relevant antibodies. Relative abundance of each band to β-actin or Erk was quantified. (C) MPO activities in colorectal tissues of each group of mice were determined. Value are presented as the mean ± SE. Means with different letters are significantly different at p < 0.05 by Duncan's multiple range test. Control, group received water without DSS; DSS, group received 3% DSS in drinking water for 5 days; DSS+MT, group received 3% DSS and oral administration with MT water extracts (5 g/kg/day) for 5 days; DSS, dextran sulfate sodium; MT, mulberry twig; MPO, myeloperoxidase
Click for larger image

Notes

This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIP) (No. NRF-2015R1C1A2A01054514).

References
1. Abraham C, Cho JH. Inflammatory bowel disease. N Engl J Med 2009;361(21):2066–2078.
2. Eaden JA, Abrams KR, Mayberry JF. The risk of colorectal cancer in ulcerative colitis: a meta-analysis. Gut 2001;48(4):526–535.
3. Gillen CD, Walmsley RS, Prior P, Andrews HA, Allan RN. Ulcerative colitis and Crohn's disease: a comparison of the colorectal cancer risk in extensive colitis. Gut 1994;35(11):1590–1592.
4. van Hogezand RA, Eichhorn RF, Choudry A, Veenendaal RA, Lamers CB. Malignancies in inflammatory bowel disease: fact or fiction. Scand J Gastroenterol Suppl 2002;37(236):48–53.
5. Yang SK, Loftus EV Jr, Sandborn WJ. Epidemiology of inflammatory bowel disease in Asia. Inflamm Bowel Dis 2001;7(3):260–270.
6. Ng WK, Wong SH, Ng SC. Changing epidemiological trends of inflammatory bowel disease in Asia. Intest Res 2016;14(2):111–119.
7. Yang DH, Yang SK. Trends in the incidence of ulcerative colitis in Korea. Korean J Med 2009;76(6):637–642.
8. Medzhitov R, Janeway CA Jr. Innate immunity: impact on the adaptive immune response. Curr Opin Immunol 1997;9(1):4–9.
9. Coussens LM, Werb Z. Inflammation and cancer. Nature 2002;420(6917):860–867.
10. Jang YJ, Leem HH, Jeon YH, Lee DH, Choi SW. Isolation and identification of α-glucosidase inhibitors from morus root bark. J Korean Soc Food Sci Nutr 2015;44(7):1090–1099.
11. Choi SW, Jang YJ, Lee YJ, Leem HH, Kim EO. Analysis of functional constituents in mulberry (Morus alba L.) twigs by different cultivars, producing areas, and heat processings. Prev Nutr Food Sci 2013;18(4):256–262.
12. Zhang Z, Shi L. Anti-inflammatory and analgesic properties of cis-mulberroside A from Ramulus mori. Fitoterapia 2010;81(3):214–218.
13. Chung KO, Kim BY, Lee MH, Kim YR, Chung HY, Park JH, et al. In-vitro and in-vivo anti-inflammatory effect of oxyresveratrol from Morus alba L. J Pharm Pharmacol 2003;55(12):1695–1700.
14. Cooper HS, Murthy SN, Shah RS, Sedergran DJ. Clinicopathologic study of dextran sulfate sodium experimental murine colitis. Lab Invest 1993;69(2):238–249.
15. Hendrickson BA, Gokhale R, Cho JH. Clinical aspects and pathophysiology of inflammatory bowel disease. Clin Microbiol Rev 2002;15(1):79–94.
16. Kornbluth A, Sachar DB. Practice Parameters Committee of the American College of Gastroenterology. Ulcerative colitis practice guidelines in adults: American College of Gastroenterology, Practice Parameters Committee. Am J Gastroenterol 2010;105(3):501–523.
17. Su C, Lichtenstein GR. Ulcerative colitis. In: Feldman M, Friedman LS, Brandt LJ, editors. Sleisenger and Fordtran's Gastrointestinal and Liver Disease: Pathophysiology, Diagnosis, Management. Volume 2. 8th edition. Philadelphia (PA): Saunders; 2006. pp. 2499-2548.
18. Choi CH, Moon W, Kim YS, Kim ES, Lee BI, Jung Y, et al. Second Korean guideline for the management of ulcerative colitis. Korean J Gastroenterol 2017;69(1):1–28.
19. Okayasu I, Hatakeyama S, Yamada M, Ohkusa T, Inagaki Y, Nakaya R. A novel method in the induction of reliable experimental acute and chronic ulcerative colitis in mice. Gastroenterology 1990;98(3):694–702.
20. Walsh-Reitz MM, Huang EF, Musch MW, Chang EB, Martin TE, Kartha S, et al. AMP-18 protects barrier function of colonic epithelial cells: role of tight junction proteins. Am J Physiol Gastrointest Liver Physiol 2005;289(1):G163–G171.
21. Araki Y, Sugihara H, Hattori T. In vitro effects of dextran sulfate sodium on a Caco-2 cell line and plausible mechanisms for dextran sulfate sodium-induced colitis. Oncol Rep 2006;16(6):1357–1362.
22. Johansson ME, Gustafsson JK, Sjöberg KE, Petersson J, Holm L, Sjövall H, et al. Bacteria penetrate the inner mucus layer before inflammation in the dextran sulfate colitis model. PLoS One 2010;5(8):e12238
23. Morgan ME, Zheng B, Koelink PJ, van de, Haazen LC, van Roest M, et al. New perspective on dextran sodium sulfate colitis: antigen-specific T cell development during intestinal inflammation. PLoS One 2013;8(7):e69936
24. Forbes E, Murase T, Yang M, Matthaei KI, Lee JJ, Lee NA, et al. Immunopathogenesis of experimental ulcerative colitis is mediated by eosinophil peroxidase. J Immunol 2004;172(9):5664–5675.
25. Choi IY, Lee KT, Kim MC, Kim SJ, Kim DS, Jeon YD, et al. Anti-inflammatory effects of Cheongilppong on DSS-induced ulcerative colitis in mice. Orient Pharm Exp Med 2011;11(1):35–39.
26. Scott RJ, Hall PA, Haldane JS, van Noorden S, Price Y, Lane DP, et al. A comparison of immunohistochemical markers of cell proliferation with experimentally determined growth fraction. J Pathol 1991;165(2):173–178.
27. Min HY, Chung HJ, Kim EH, Kim S, Park EJ, Lee SK. Inhibition of cell growth and potentiation of tumor necrosis factor-α (TNF-α)-induced apoptosis by a phenanthroindolizidine alkaloid antofine in human colon cancer cells. Biochem Pharmacol 2010;80(9):1356–1364.
28. Hoffmann A, Leung TH, Baltimore D. Genetic analysis of NF-κ B/Rel transcription factors defines functional specificities. EMBO J 2003;22(20):5530–5539.
29. Kinoshita T, Ito H, Miki C. Serum interleukin-6 level reflects the tumor proliferative activity in patients with colorectal carcinoma. Cancer 1999;85(12):2526–2531.
30. Janssen-Heininger YM, Poynter ME, Baeuerle PA. Recent advances towards understanding redox mechanisms in the activation of nuclear factor κB. Free Radic Biol Med 2000;28(9):1317–1327.
31. Schulze-Osthoff K, Ferrari D, Riehemann K, Wesselborg S. Regulation of NF-κ B activation by MAP kinase cascades. Immunobiology 1997;198(1-3):35–49.
32. Costa F, Mumolo MG, Ceccarelli L, Bellini M, Romano MR, Sterpi C, et al. Calprotectin is a stronger predictive marker of relapse in ulcerative colitis than in Crohn's disease. Gut 2005;54(3):364–368.