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Gut microbiome and inflammation in response to increasing intermittent hypoxia in the neonatal rat

Pediatric Research volume 97, pages 2126–2135 (2025)Cite this article

Abstract

Background

Intermittent hypoxia (IH) and oxidative stress play key roles in gut dysbiosis and inflammation. We tested the hypothesis that increasing numbers of daily IH episodes cause microbiome dysbiosis and severe gut injury.

Methods

Neonatal rats were exposed to hyperoxia (Hx), growth restriction, and IH. For IH, pups were exposed to 2–12 daily episodes from birth (P0) to postnatal day 7 (7D) or P0-P14 (14D), with or without recovery in room air (RA) until P21. Animals raised in RA from P0 to P21 served as normoxia controls. Stool was expressed from the large intestines for microbiome analysis, and tissue samples were assessed for histopathology and biomarkers of inflammation.

Results

Hx and IH caused a significant reduction in the number and diversity of organisms. The severity of gut injury and levels of inflammatory cytokines and TLR4 increased, while total glutathione (tGSH) declined, with increasing daily IH episodes. The number of organisms correlated with the villi number (p < 0.05) and tGSH depletion (p < 0.001).

Conclusions

The critical number of daily IH episodes that the newborn gut may sustain is 6, beyond which irreversible damage occurs. The immature gut is highly susceptible to IH-induced injury, and IH may contribute to pathological outcomes in the immature gut.

Impact statement

  1. 1.

    The neonatal gut at birth is highly susceptible to intermittent hypoxia (IH) injury.

  2. 2.

    IH causes gut dysbiosis, inflammation, and glutathione depletion.

  3. 3.

    The severity of gut injury worsens as a function of increasing daily IH episodes.

  4. 4.

    The critical number of daily IH episodes that the newborn gut may sustain is 6, beyond which irreversible damage occurs.

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Fig. 1: Representative H&E stained images of large intestines from neonatal rats exposed to room air (RA, top panels), hyperoxia (50% O2; middle panels), or intermittent hypoxia (IH, lower panels).
Fig. 2: Changes in the relative proportion of the most abundant microorganisms in response to increasing daily neonatal IH episodes.
Fig. 3: Effect of increasing daily IH episodes on IL-1β, (panels a and d), IL-6 (panels b and e), and TNFα (panels c and f) in the large intestine homogenates.
Fig. 4: Effect of increasing daily IH episodes on TLR4, TGFβ, and total glutathione (tGSH) levels in the large intestine homogenates.
Fig. 5: Representative TLR-4 immunostaining.

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

This work was made possible through the Eunice Kennedy Shriver National Institute of Child Health & Human Development Grant #U54HD071594.

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Authors and Affiliations

  1. Department of Pediatrics, Division of Neonatal-Perinatal Medicine, State University of New York, Downstate Health Sciences University, Brooklyn, NY, USA

    Magdalena Latkowska, Charles L. Cai, Jacob V. Aranda & Kay D. Beharry

  2. Department of Pediatrics, Downstate Medical Center, State University of New York, Brooklyn, NY, USA

    Marina Mitrou

  3. SUNY Eye Institute, Brooklyn, NY, USA

    Matthew Marcelino, Jacob V. Aranda & Kay D. Beharry

  4. Department of Ophthalmology, Downstate Medical Center, State University of New York, Brooklyn, NY, USA

    Jacob V. Aranda & Kay D. Beharry

Authors
  1. Magdalena Latkowska
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  2. Charles L. Cai
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  4. Matthew Marcelino
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  5. Jacob V. Aranda
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  6. Kay D. Beharry
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Contributions

All authors have made substantial contributions to the conception (M.L., J.V.A., K.D.B.), methodology and design of the work (M.L., J.V.A., K.D.B.), investigation (M.L., C.L.C., M.M., M.M., K.D.B.), acquisition and formal analysis (M.L., C.L.C., M.M., M.M., K.D.B.), validation (M.L., J.V.A., K.D.B.), data curation (M.L., K.D.B.), interpretation of the data (M.L., J.V.A., K.D.B.), visualization (M.L., C.L.C., M.M., M.M., K.D.B.), original draft (M.L.), and review and approval of the submitted version (M.L., C.L.C., M.M., M.M., J.V.A., K.D.B.), supervision (J.V.A., K.D.B.), project administration (J.V.A., K.D.B.), funding acquisition (J.V.A.).

Corresponding author

Correspondence to Kay D. Beharry.

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Latkowska, M., Cai, C.L., Mitrou, M. et al. Gut microbiome and inflammation in response to increasing intermittent hypoxia in the neonatal rat. Pediatr Res 97, 2126–2135 (2025). https://doi.org/10.1038/s41390-024-03569-7

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