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Betaine improves hyperoxic lung injury through downregulating pulmonary macrophage pyroptosis in newborn mice

Pediatric Research (2025)Cite this article

Abstract

Background

Bronchopulmonary dysplasia (BPD) is a common chronic respiratory disease in preterm infants. NLRP3-mediated macrophage pyroptosis plays a crucial role in the pathogenesis of BPD. Recent evidence suggests that betaine has anti-inflammatory and antioxidant functions. This study aimed to investigate the effect of betaine on pulmonary macrophage pyroptosis in BPD.

Methods

Newborn mice were exposed to either hyperoxia (90%) or room air shortly after birth and treated subcutaneously with betaine daily for 14 days. Lung development, the expression of macrophage pyrolysis-associated proteins and the phospho-forkhead box O1 (p-FOXO1) were evaluated. In vitro, the effect of betaine on the expression of p-FOXO1 was assessed in RAW264.7 macrophages exposed to either 90% oxygen or 21% oxygen, with okadaic acid (OA) as phosphatase inhibitor.

Results

Hyperoxia induced macrophage pyroptosis and impaired lung development in newborn mice. Betaine inhibited p-FOXO1 expression and NLRP3-mediated pyroptosis in pulmonary and promoted lung development in the hyperoxia-exposed mice. In vitro, betaine suppressed FOXO1 phosphorylation and NLRP3-mediated pyroptosis under 90% oxygen in RAW264.7 cells, and OA administration reversed these effects.

Conclusion

Betaine may reduce the expression of inflammatory cytokines, downregulate macrophage pyroptosis by inhibiting the phosphorylation of FOXO1, and improve lung development in BPD.

Impact

  • Our previous research showed that plasma betaine levels at 36 weeks postmenstrual age (PMA) were significantly lower in preterm infants with BPD compared to those without BPD.

  • Betaine has anti-inflammatory and antioxidant functions. However, its role in pulmonary macrophage pyroptosis in BPD remains unknown.

  • The study shows that betaine may downregulate macrophage pyroptosis by inhibiting FOXO1 phosphorylation, alleviate lung inflammation, and improve lung development in BPD.

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Fig. 1: Betaine attenuates lung impairment in newborn mice exposed to hyperoxia.
Fig. 2: Betaine downregulates the expression of inflammatory factors and pyroptosis in newborn mice exposed to hyperoxia on P14.
Fig. 3: Betaine downregulates macrophage pyroptosis in the pulmonary macrophage in mice exposed to hyperoxia.
Fig. 4: Betaine inhibits FOXO1 phosphorylation in the pulmonary macrophages in mice exposed to hyperoxia.
Fig. 5: Betaine alleviates pyroptosis in RAW264.7 macrophages exposed to 90% oxygen for 48 hours.
Fig. 6: Betaine decreases the expression of p-FOXO1 and NLRP3 in RAW264.7 macrophages exposed to 90% oxygen for 48 h.

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Data availability

The datasets used or analyzed during the current study are available from the corresponding author upon reasonable request.

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Funding

This work was partially supported by National Natural Science Foundation of China (No. 82471744) and Shanghai Municipal Health Commission (No. 202340047).

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Author notes

  1. These authors contributed equally: Jufeng Zhang, Lin Zhou.

Authors and Affiliations

  1. Department of Neonatology, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China

    Jufeng Zhang, Lin Zhou, Hui Xu, You You, Siyi Xia & Hongping Xia

Authors
  1. Jufeng Zhang
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  2. Lin Zhou
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  3. Hui Xu
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  4. You You
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  5. Siyi Xia
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  6. Hongping Xia
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Contributions

Jufeng Zhang contributed to the animal study, molecular biology examination, and data analyses; Lin Zhou contributed to data analyses; Hui Xu contributed to in vitro study; You You and Siyi Xia contributed to immunofluorescence staining and Western blotting; Hongping Xia contributed to the design of the study and revised the article for important intellectual content; all authors have read and approved the final version to be published.

Corresponding author

Correspondence to Hongping Xia.

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The authors declare no competing interests.

Ethics approval

The study was reviewed and approved by Ethics Committee of Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine (XHEC-F-2024-025). All methods were carried out in accordance with relevant guidelines and regulations.

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Zhang, J., Zhou, L., Xu, H. et al. Betaine improves hyperoxic lung injury through downregulating pulmonary macrophage pyroptosis in newborn mice. Pediatr Res (2025). https://doi.org/10.1038/s41390-025-04364-8

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