J Nutr Health.  2019 Apr;52(2):139-148. 10.4163/jnh.2019.52.2.139.

Anti-inflammatory effects of mulberry twig extracts on dextran sulfate sodium-induced colitis mouse model

• Cui X
• Kim E

• KMID: 2444937
• DOI: http://doi.org/10.4163/jnh.2019.52.2.139

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.

Keyword

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

MeSH Terms

Animals
Colitis*
Colitis, Ulcerative
Colon
Colorectal Neoplasms
Dextran Sulfate*
Dextrans*
Drinking Water
Far East
Humans
Inflammatory Bowel Diseases
Interleukin-6
Male
Mice*
Mice, Inbred ICR
Morus*
Peroxidase
Plasma
Risk Factors
RNA, Messenger
Water
Dextran Sulfate
Dextrans
Drinking Water
Interleukin-6
Peroxidase
RNA, Messenger
Water

Figure

• 

  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

• 

  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

• 

  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

• 

  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

• 

  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

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