Nutr Res Pract.  2018 Apr;12(2):101-109. 10.4162/nrp.2018.12.2.101.

Effect of vitamin C on azoxymethane (AOM)/dextran sulfate sodium (DSS)-induced colitis-associated early colon cancer in mice

• Jeon HJ
• Yeom Y
• Kim YS
• Kim E
• Shin JH
• Seok PR
• Woo MJ
• Kim Y

• KMID: 2407806
• DOI: http://doi.org/10.4162/nrp.2018.12.2.101

Abstract

BACKGROUND/OBJECTIVES
The objective of this study was to investigate the effects of vitamin C on inflammation, tumor development, and dysbiosis of intestinal microbiota in an azoxymethane (AOM)/dextran sulfate sodium (DSS)-induced inflammation-associated early colon cancer mouse model.
MATERIALS/METHODS
Male BALB/c mice were injected intraperitoneally with AOM [10 mg/kg body weight (b.w)] and given two 7-d cycles of 2% DSS drinking water with a 14 d inter-cycle interval. Vitamin C (60 mg/kg b.w. and 120 mg/kg b.w.) was supplemented by gavage for 5 weeks starting 2 d after the AOM injection.
RESULTS
The vitamin C treatment suppressed inflammatory morbidity, as reflected by disease activity index (DAI) in recovery phase and inhibited shortening of the colon, and reduced histological damage. In addition, vitamin C supplementation suppressed mRNA levels of pro-inflammatory mediators and cytokines, including cyclooxygenase-2, microsomal prostaglandin E synthase-2, tumor necrosis factor-α, Interleukin (IL)-1β, and IL-6, and reduced expression of the proliferation marker, proliferating cell nuclear antigen, compared to observations of AOM/DSS animals. Although the microbial composition did not differ significantly between the groups, administration of vitamin C improved the level of inflammation-related Lactococcus and JQ084893 to control levels.
CONCLUSION
Vitamin C treatment provided moderate suppression of inflammation, proliferation, and certain inflammation-related dysbiosis in a murine model of colitis associated-early colon cancer. These findings support that vitamin C supplementation can benefit colonic health. Long-term clinical studies with various doses of vitamin C are warranted.

Keyword

Vitamin C; colitis; inflammation; colonic neoplasm; microbiota

MeSH Terms

Animals
Ascorbic Acid*
Azoxymethane*
Body Weight
Colitis
Colon*
Colonic Neoplasms*
Cyclooxygenase 2
Cytokines
Drinking Water
Dysbiosis
Gastrointestinal Microbiome
Humans
Inflammation
Interleukin-6
Interleukins
Lactococcus
Male
Mice*
Microbiota
Necrosis
Proliferating Cell Nuclear Antigen
RNA, Messenger
Sodium*
Vitamins*
Ascorbic Acid
Azoxymethane
Cyclooxygenase 2
Cytokines
Drinking Water
Interleukin-6
Interleukins
Proliferating Cell Nuclear Antigen
RNA, Messenger
Sodium
Vitamins

Figure

• 

  Fig. 1 Effects of vitamin C supplementation on DAI score. DAI scores reflect weight loss, stool consistency, and fecal bleeding. (A) Bi-daily group DAI scores. (B) Analysis of day 16th, DAI scores among the Control, CC, V60, and V120 groups. Mean ± SEM are shown (Control, n = 9; CC, n = 12; V60 and V120, n = 11 per group). One-way ANOVAs and Newman-Keuls' post hoc tests were performed (P<0.05). CC, AOM/DSS-induced colon cancer; V60, AOM/DSS + 60 mg/kg body weight (b.w.) of vitamin C; V120, AOM/DSS + 120 mg/kg b.w. of vitamin C.

• 

  Fig. 2 Effects of vitamin C supplementation on colon length and histology in colitis-associated colon cancer. (A) Colon length was measured and compared among four groups. Mean ± SEM are shown (Control, n = 9; CC, n = 12; V60 and V120, n = 11 per group). One-way ANOVAs and Newman-Keuls' post hoc tests were performed (P<0.05). (B) Representative histologic sections of distal colon from among four groups. Glandular epithelium destruction (arrow head), neutrophil infiltration (arrow) or submucosa extension (S) were shown. HE stain, bar = 100 µm. Magnification, upper 100x, lower 200x CC, AOM/DSS-induced colon cancer; V60, AOM/DSS + 60 mg/kg body weight (b.w.) of vitamin C; V120, AOM/DSS + 120 mg/kg b.w. of vitamin C.

• 

  Fig. 3 Effect of vitamin C supplementation on proinflammatory mediators and cytokines in middle and distal colon. (A) mRNA expressions of COX-2 and mPGES-2 (B) TNF-α, IL-1β, and IL-6 in middle colon (a) and in the distal colon (b) were analyzed using real-time PCR. β-actin and GAPDH were used as the loading control. Mean ± SEM are shown (Control, n = 9; CC, n = 12; V60 and V120, n = 11 per group). One-way ANOVAs and Newman-Keuls' post hoc tests were performed (P<0.05). CC, AOM/DSS-induced colon cancer; V60, AOM/DSS + 60 mg/kg body weight (b.w.) of vitamin C; V120, AOM/DSS + 120 mg/kg b.w. of vitamin C.

• 

  Fig. 4 The effects of vitamin C on colon tumor number and mRNA expression of PCNA. (A) Number of colon tumor were counted with naked eye when sacrificing the mice. (B) mRNA expressions of PCNA was analyzed in the middle colon (a) and in the distal colon (b) using real-time PCR. β-actin and GAPDH were used as the loading control. Mean ± SEM are shown (Control, n = 9; CC, n = 12; V60 and V120, n = 11 per group). One-way ANOVAs and Newman-Keuls' post hoc tests were performed (P<0.05). CC, AOM/DSS-induced colon cancer; V60, AOM/DSS + 60 mg/kg body weight (b.w.) of vitamin C; V120, AOM/DSS + 120 mg/kg b.w. of vitamin C.

• 

  Fig. 5 Effects of vitamin C on gut microbiome taxonomy. (A) Composition of gut microbacteria at the phylum level. (a) Changes in the proportion of Bacteroidetes and Firmicutes phyla. (b) Relative abundance of Bacteridetes/Firmicutes across the Control, CC, and V60 groups. (B) Relative abundance of genus Lactoccocus. (C) Relative abundance of JQ084893 species. Mean ± SEM are shown (n = 6 per group). One-way ANOVAs and Newman-Keuls' post hoc tests were used (P<0.05). CC, AOM/DSS-induced colon cancer; V60, AOM/DSS + 60 mg/kg body weight (b.w.) of vitamin C.

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