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Dietary riboflavin deficiency induces ariboflavinosis and esophageal epithelial atrophy in association with modification of gut microbiota in rats

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European Journal of Nutrition Aims and scope Submit manuscript

Abstract

Purpose

Riboflavin deficiency causes ariboflavinosis, a common nutritional deficiency disease. The purpose of this study is to investigate the effects of riboflavin deficiency on the important internal organs and its potential mechanisms.

Methods

Experiment 1, male F344 rats were randomly assigned to R6 (normal riboflavin, 6 mg/kg) and R0 (riboflavin-deficient, 0 mg/kg) groups. Experiment 2 rats were assigned to R6, R0.6 (0.6 mg/kg) and R0.06 (0.06 mg/kg) groups. Experiment 3 rats were assigned to R6 and R0 → R6 (riboflavin replenishment) groups. Bacterial communities were analyzed based on 16S rRNA gene sequencing.

Results

Riboflavin deficiency induced ariboflavinosis (R0.06 46.7%; R0 72%) and esophageal epithelial atrophy (R0.06 40%; R0 44%) in rats, while the R6 group did not display symptoms (P < 0.001, respectively). Esophageal epithelial atrophy occurred simultaneously (R0.06 66.7%; R0 63.6%) with ariboflavinosis or appeared alone (R0.06 33.3%; R0 36.4%). Esophagus is the most vulnerable internal organ. Riboflavin deficiency followed by replenishment (R0 → R6) was effective in treating ariboflavinosis (83.3% vs. 0%, P < 0.001) and esophageal epithelial atrophy (66.7% vs. 20%, P = 0.17). Riboflavin deficiency modulated gut microbiota composition. The several key genera (Romboutsia, Turicibacter and Clostridium sensu stricto 1) were strongly correlated with ariboflavinosis and esophageal epithelial atrophy (P < 0.01 or P < 0.05). The potential mechanism is that gut microbiota affects body's xenobiotic biodegradation and metabolism, and genomic instability.

Conclusions

Riboflavin deficiency induces ariboflavinosis and esophageal epithelial atrophy by modulating the gut microbiota, and offers new Queryinsight into riboflavin deficiency and esophageal lesions.

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Data availability

All data are available in the manuscript or upon request to the authors.

Abbreviations

Expt.:

Experiment

EIN:

Esophageal intraepithelial neoplasia

FLASH:

Fast length adjustment of short reads

g:

Gram

nM:

Nmol/L

OTUs:

Operational taxonomic units

PCoA:

Principal coordinates analysis

PICRUSt:

Phylogenetic Investigation of Communities by Reconstruction of Unobserved States

QIIME2:

Quantitative insights into microbial ecology 2

R6 :

Riboflavin 6 mg/kg

R0 :

Riboflavin 0 mg/kg

R0.6 :

Riboflavin 0.6 mg/kg

R0.06 :

Riboflavin 0.06 mg/kg

R0 → R6 :

Riboflavin 0 mg/kg for weeks 0–20 and then 6 mg/kg for weeks 21–35

SCFAs:

Short-chain fatty acids

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Acknowledgements

We thank Dr. Stanley Li Lin, Department of Cell Biology and Genetics, Shantou University Medical College, for assistance in revising the manuscript. We are grateful for assistance from the Central Laboratory at Shantou University Medical College, including Prof. Wen-Hong Luo, for obtaining the riboflavin concentration data by HPLC.

Funding

This work was supported by Grants from the Natural Science Foundation of China-Guangdong Joint Fund (No. U1301227), the National Cohort of Esophageal Cancer of China (2016YFC0901400).

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Authors and Affiliations

  1. Key Laboratory of Molecular Biology for High Cancer Incidence Coastal Chaoshan Area, Shantou University Medical College, Shantou, 515041, China

    Feng Pan, Ye Chen, Lin Long, Xuan Wang, Pei-Tong Zhuang, En-Min Li & Li-Yan Xu

  2. Department of Biochemistry and Molecular Biology, Shantou University Medical College, No.22, Xinling Road, Shantou, 515041, Guangdong, China

    Feng Pan, Lin Long, Xuan Wang, Pei-Tong Zhuang & En-Min Li

  3. Department of Experimental Animal Center, Shantou University Medical College, Shantou, 515041, China

    Ling-Li Zhang

  4. Bioanalytical Laboratory, Shantou University Medical College, Shantou, 515041, China

    Hong-Jun Luo

  5. Institute of Oncologic Pathology, Shantou University Medical College, No.22, Xinling Road, Shantou, 515041, Guangdong, China

    Ye Chen & Li-Yan Xu

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  1. Feng Pan
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Contributions

LYX and EML planned the studies. FP, LLZ, HJL, YC, LL, XW, and PTZ performed the experiments. FP held the responsibility for all data integrity and data analysis. LYX and EML conducted the whole research. LYX had primary responsibility for the final content. All authors have read and approved the final manuscript.

Corresponding authors

Correspondence to En-Min Li or Li-Yan Xu.

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Conflict of interest

The authors declare no conflict of interest.

Ethical approval and consent to participate

All experimental procedures administered on animals were approved by the Institutional Animal Care and Use Committee of Shantou University.

Electronic supplementary material

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394_2020_2283_MOESM1_ESM.tif

Supplemental Fig. 1. Riboflavin deficiency induces low body weight (A) and ariboflavinosis (B) in rats. (C) Riboflavin replenishment is effective in treating ariboflavinosis (TIF 14938 kb)

394_2020_2283_MOESM2_ESM.tif

Supplemental Fig. 2. Representative pictures of hematoxylin and eosin-stained gastrointestinal tissue (A) and other major internal tissues (B). Scale bars, 50 μm (TIF 23814 kb)

Supplemental Fig. 3. Rarefaction curves (A) and Shannon-Wiener curves (B) (TIF 1564 kb)

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Pan, F., Zhang, LL., Luo, HJ. et al. Dietary riboflavin deficiency induces ariboflavinosis and esophageal epithelial atrophy in association with modification of gut microbiota in rats. Eur J Nutr 60, 807–820 (2021). https://doi.org/10.1007/s00394-020-02283-4

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