Intermittent fasting prompted recovery from dextran sulfate sodium-induced colitis in mice - PubMed (original) (raw)
. 2017 Sep;61(2):100-107.
doi: 10.3164/jcbn.17-9. Epub 2017 Jul 28.
Affiliations
- PMID: 28955126
- PMCID: PMC5612824
- DOI: 10.3164/jcbn.17-9
Intermittent fasting prompted recovery from dextran sulfate sodium-induced colitis in mice
Toshihiko Okada et al. J Clin Biochem Nutr. 2017 Sep.
Abstract
Fasting-refeeding in mice induces transient hyperproliferation of colonic epithelial cells, which is dependent on the lactate produced as a metabolite of commensal bacteria. We attempted to manipulate colonic epithelial cell turnover with intermittent fasting to prompt recovery from acute colitis. Acute colitis was induced in C57BL/6 mice by administration of dextran sulfate sodium in the drinking water for 5 days. From day 6, mice were fasted for 36 h and refed normal bait, glucose powder, or lactylated high-amylose starch. On day 9, colon tissues were subjected to analysis of histology and cytokine expression. The effect of lactate on the proliferation of colonocytes was assessed by enema in vivo and primary culture in vitro. Intermittent fasting resulted in restored colonic crypts and less expression of interleukin-1β and interleukin-17 in the colon than in mice fed ad libitum. Administration of lactate in the colon at refeeding time by enema or by feeding lactylated high-amylose starch increased the number of regenerating crypts. Addition of lactate but not butyrate or acetate supported colony formation of colonocytes in vitro. In conclusion, intermittent fasting in the resolution phase of acute colitis resulted in better recovery of epithelial cells and reduced inflammation.
Keywords: NDRG3; colitis; interleukin-17; interleukin-1β; lactate.
Conflict of interest statement
No potential conflicts of interest were disclosed.
Figures
Fig. 1
Intermittent fasting restored crypts and decreased cytokine production in colitis. (A) Body weight changes. Mice were fed CE-2 ad libitum without intermittent fasting (Ad lib); mice were fed CE-2, fasted at the indicated period and refed CE-2 (F + CE2); mice were fed CE-2, fasted and fed glucose and salt (F + Glu). Data are shown as mean ± SD of 6 mice for each group. A statistically significant difference the between F + CE2 and F + Glu groups (p<0.01) was seen on day 9 (two-way ANOVA with Bonferroni multiple comparisons). (B) Typical histological findings of the colon on day 9. Frozen sections were stained with hematoxylin & eosin. (C) Numbers of crypts were determined from the images as panel (B). Each dot represents an individual mouse and the short bar indicates the mean value (*p<0.02, Mann–Whitney U test). (D) Levels of cytokine transcripts in the colonic tissue on day 9. Data are shown as relative expression to naïve mice without induction of colitis. Each dot represents an individual mouse, and the short bar indicates the mean value. (*p<0.02, Mann–Whitney test). DSS, dextran sulfate sodium; IL, interleukin; TNF, tumor necrosis factor.
Fig. 2
Delivery of lactate into the colon ameliorated colitis. (A) Body weight changes. Mice were fed CE-2, fasted at the indicated period and refed lactylated high-amylose maize starches (LHA, n = 7) or control high-amylose maize starches (HA, n = 6). Data are shown as mean ± SD. There was no statistically significant difference (two-way ANOVA). (B) Numbers of crypts were determined on day 9 from the hematoxylin & eosin (H&E)-stained frozen sections. Each dot represents an individual mouse, and the short bar indicates the mean value. *p<0.05 (Mann–Whitney U test). (C) Total ulcer length determined on day 9 from the H&E stained frozen sections prepared from the rolled whole colon. Each dot represents an individual mouse, and the short bar indicates the mean value. *p<0.02 (Mann–Whitney test). (D) Levels of cytokine transcripts in the colonic tissue on day 9. Data are shown as relative expression to naïve mice without induction of colitis. Each dot represents an individual mouse, and the short bar indicates the mean value. (*p<0.05, Mann–Whitney test). DSS, dextran sulfate sodium; IL, interleukin; TNF, tumor necrosis factor.
Fig. 3
Direct effect of lactate on epithelial cells in vivo and in vitro. (A) DSS-treated mice were fasted for 36 h from day 6–8 and refed with glucose (Glu) powder with enema of organic acids or saline as a vehicle. Enema was started at the timing of refeeding and repeated every 3 h 5 times. Numbers of crypts were determined as in Fig. 1C. Data are shown as mean ± SD of 3 mice for each experimental condition. *p<0.05 (One way ANOVA with Tukey’s multiple comparison test). (B) Colonocytes from fasted (36 h) mice were cultured with or without lactate, and from day 4, number of colonies/well was counted every day. A typical colony is shown in the upper panel. Data are shown as mean ± SD of triplicated culture for each experimental condition. *p<0.05 (Two-way RM ANOVA with Bonferroni multiple comparisons). (C) Colonocytes from mice fed ad libitum (ad lib) or fasted for 36 h were cultured with Lactate (Lac), butyrate (But), acetate (Ace), or vehicle (No). Number of colonies/well on day 7 (left) and 13 (right) are shown. Data are shown as mean ± SD of triplicated culture for each experimental condition. *p<0.05 (one-way ANOVA with Tukey’s multiple comparison test). (D) Western blotting of the total colon homogenate from mice on day 9, with intermittent fasting (fasting-refed) or without (ad lib) for detection of Hif1α and NDRG3. Relative expression levels of NDRG3 to tubulin were quantified using samples from 4 mice for each condition and summarized in the graph (mean ± SD, *p<0.05, Mann–Whitney U test).
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