Figure 2.
DMB decreased high-choline-diet–produced TMAO and reduced tissue damage in choline-fed GVHD mice. (A) Experimental scheme of BMT mice and GVHD mice protocol. BALB/c recipients were fed with DMB, choline-containing diet, or choline+DMB from day −14 and, thereafter, lethally irradiated on day −1, and transfused with C57BL/6 donor cells on day 0. The data were collected on day 14 except for the survival curve analysis on day 50. (B) Serum TMAO concentration was determined in GVHD mice fed with DMB, choline, or choline and DMB diet. Data were pooled from 3 independent experiments. N = 7 in each group. **P < .01. (C) Kaplan-Meier survival curve was determined in GVHD mice with vehicle (n = 25), DMB (n = 20), choline (n = 25), or choline+DMB diet (n = 25). Graph represents pooled data from 2 (DMB) or 3 (vehicle, choline, choline+DMB) independent experiments. **P < .01 (TCD-BM+T vs TCD-BM+T+choline). ##P < .01 (TCD-BM+T+choline vs TCD-BM+T+choline+DMB). (D) GVHD score was determined in GVHD mice treated with vehicle (n = 25), DMB (n = 20), choline (n = 25), or choline+DMB (n = 25). Graph represents pooled data from 2 (DMB) or 3 (vehicle, choline, choline+DMB) independent experiments. *P < .05, **P < .01 (TCD-BM+T vs TCD-BM+T+choline). #P < .05, ##P < .01 (TCD-BM+T+choline vs TCD-BM+T+choline+DMB). (E) Representative H&E staining of ileum, colon, skin, and liver tissues from GVHD mice fed with choline, DMB, or choline and DMB diet. Scale bar, 200 μm. (F) Histologic analyses of paraffin-sectioned ileum, colon, skin, and liver tissues from GVHD mice were performed on day 14. *P < .05, **P < .01. Data were pooled from 3 independent experiments. N = 10 in each group. (G) Bacterial diversity of fecal microbiota from GVHD mice with high-choline diet or control diet was analyzed. N = 6 in each group. (H) Principal coordinate analysis distances of the fecal microbiota communities between high-choline diet and chow diet in GVHD mice. N = 6 in each group. (I) Linear discriminant analysis coupled with effect size measurements to identify gut microbial taxa in GVHD mice fed with high-choline diet or control diet. N = 6 in each group. ns, not significant; PC, principal component.

DMB decreased high-choline-diet–produced TMAO and reduced tissue damage in choline-fed GVHD mice. (A) Experimental scheme of BMT mice and GVHD mice protocol. BALB/c recipients were fed with DMB, choline-containing diet, or choline+DMB from day −14 and, thereafter, lethally irradiated on day −1, and transfused with C57BL/6 donor cells on day 0. The data were collected on day 14 except for the survival curve analysis on day 50. (B) Serum TMAO concentration was determined in GVHD mice fed with DMB, choline, or choline and DMB diet. Data were pooled from 3 independent experiments. N = 7 in each group. **P < .01. (C) Kaplan-Meier survival curve was determined in GVHD mice with vehicle (n = 25), DMB (n = 20), choline (n = 25), or choline+DMB diet (n = 25). Graph represents pooled data from 2 (DMB) or 3 (vehicle, choline, choline+DMB) independent experiments. **P < .01 (TCD-BM+T vs TCD-BM+T+choline). ##P < .01 (TCD-BM+T+choline vs TCD-BM+T+choline+DMB). (D) GVHD score was determined in GVHD mice treated with vehicle (n = 25), DMB (n = 20), choline (n = 25), or choline+DMB (n = 25). Graph represents pooled data from 2 (DMB) or 3 (vehicle, choline, choline+DMB) independent experiments. *P < .05, **P < .01 (TCD-BM+T vs TCD-BM+T+choline). #P < .05, ##P < .01 (TCD-BM+T+choline vs TCD-BM+T+choline+DMB). (E) Representative H&E staining of ileum, colon, skin, and liver tissues from GVHD mice fed with choline, DMB, or choline and DMB diet. Scale bar, 200 μm. (F) Histologic analyses of paraffin-sectioned ileum, colon, skin, and liver tissues from GVHD mice were performed on day 14. *P < .05, **P < .01. Data were pooled from 3 independent experiments. N = 10 in each group. (G) Bacterial diversity of fecal microbiota from GVHD mice with high-choline diet or control diet was analyzed. N = 6 in each group. (H) Principal coordinate analysis distances of the fecal microbiota communities between high-choline diet and chow diet in GVHD mice. N = 6 in each group. (I) Linear discriminant analysis coupled with effect size measurements to identify gut microbial taxa in GVHD mice fed with high-choline diet or control diet. N = 6 in each group. ns, not significant; PC, principal component.

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