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Butyrate protects the intestinal barrier by upregulating Fut2 expression via MEK4-JNK signaling pathway activation

Pediatric Research (2024)Cite this article

Abstract

Background

Necrotizing enterocolitis (NEC) is a severe gastrointestinal inflammatory disease in neonates. Fucosyltransferase 2 (Fut2) regulates intestinal epithelial cell fucosylation. In this study, we aimed to investigate butyrate-mediated upregulation of Fut2 expression and the underlying mechanisms.

Methods

In vivo and in vitro models were established. SP600125 was used to inhibit the MEK4-JNK pathway, and anisomycin was used to activate the MEK4-JNK pathway. Fut2, occludin, and ZO-1 expressions were assessed. Furthermore, intestinal permeability was analyzed by FITC-Dextran. The expression of proteins in the MEK-4-JNK pathway was examined by western blotting.

Results

In vivo, the addition of exogenous butyrate notably upregulated Fut2, occludin, and ZO-1 expressions and reduced intestinal permeability in mice with NEC. Butyrate may increase the phosphorylation of MEK4, JNK, and c-jun, which are key components of the MEK4-JNK pathway. Additionally, SP600125 inhibited their phosphorylation, which was reversed by anisomycin treatment. In vitro, butyrate substantially increased occludin and ZO-1 expressions. Butyrate considerably increased Fut2 expression and markedly upregulated p-MEK4, p-JNK, and p-c-jun expressions. SP600125 administration decreased their expressions, while anisomycin administration increased their expressions.

Conclusion

Butyrate upregulated Fut2 expression via activation of the MEK4-JNK pathway, improved intestinal barrier integrity, and protected neonatal mice from NEC.

Impact

  • We found that exogenous butyrate could improve intestinal barrier integrity and protect against NEC in neonatal mice.

  • Our data showed that exogenous butyrate supplementation upregulated Fut2 expression by activating the MEK4-JNK pathway.

  • Our study provides novel insights into the pathogenesis of NEC, thereby laying an experimental foundation for future clinical research on the use of butyrate in NEC treatment.

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Fig. 1: Butyrate reduced intestinal injury in mice with NEC.
Fig. 2: IEC-6 cells were treated with different concentrations of butyrate for 16 h. Cell viability was determined using a CCK-8 assay.
Fig. 3: Effects of butyrate on the intestinal epithelial barrier integrity in vivo and in vitro.
Fig. 4: Effects of butyrate on the expression of Fut2 in vivo and in vitro.
Fig. 5: Effects of butyrate on GPR41 and the MEK4-JNK pathway in vivo and in vitro.

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Data availability

The datasets generated for this study are available from the corresponding author upon request.

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Funding

Funding

This work was supported by the Natural Science Foundation of Chongqing municipality (cstc2021jcyj-msxmX0063), the Joint Medical Research Project of Chongqing Science and Technology Commission (2021MSXM206,2022MSXM039), and the Project of Nestle Health Science (KY210030, China).

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Author notes

  1. These authors contributed equally: Dan-Dan Zhang, Zhao-Xi Huang.

Authors and Affiliations

  1. Department of Neonatology Children’s Hospital of Chongqing Medical University, National Clinical Research Center for Child Health and Disorders, Ministry of Education Key Laboratory of Child Development and Disorders, Chongqing Key Laboratory of Child Rare Disease in infection and Immunity, Chongqing, China

    Dan-Dan Zhang, Zhao-Xi Huang, Xiao-Chen Liu, Yu He, Qing Ai, Lu-Quan Li & Lei Bao

  2. Department of neonatology, Jiangxi Hospital Affiliated to Children’s Hospital of Chongqing Medical University, Children’s Medical Center of Jiangxi, Jiangxi, 330103, China

    Xiang-Ping Ding

  3. Guang ‘an maternal and Child Health and Family Planning Service Center, Sichuan, China

    Ling Li

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Contributions

All the authors made substantial contributions to the study. D.-D.Z. and Z.-X.H. led the experiments. D.-D.Z., Z.-X.H., X.-C.L. and X.-P.D. conceived and conducted the experiments and analyzed the data. L.L., Q.A., Y.H., L.B. and L.-Q.L. provided resources and equipment. D.-D.Z. wrote the original draft. X.-C.L. and X.-P.D. reviewed and edited the article. L.B. and L.-Q.L. contributed to the critical revision and final approval of the manuscript. All authors contributed to the article and approved the submitted version. D.-D.Z. and Z.-X. Huang contributed equally to this work and share first authorship.

Corresponding authors

Correspondence to Lu-Quan Li or Lei Bao.

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Zhang, DD., Huang, ZX., Liu, XC. et al. Butyrate protects the intestinal barrier by upregulating Fut2 expression via MEK4-JNK signaling pathway activation. Pediatr Res (2024). https://doi.org/10.1038/s41390-024-03419-6

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  • DOI: https://doi.org/10.1038/s41390-024-03419-6

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