ALAS2 Prevents Neonatal Necrotizing Enterocolitis by Improving Ferroptosis in Intestinal Epithelial Cells Through Inhibition of Oxidative Stress

Q: What role does ALAS2 play in neonatal necrotizing enterocolitis (NEC)?

ALAS2 expression is significantly downregulated in NEC models and is linked to oxidative stress and ferroptosis activation. Restoring ALAS2 via lentiviral overexpression mitigates injury by blocking oxidative stress-driven ferroptosis in intestinal epithelial cells.

Q: How was the NEC model induced and what changes were observed?

A neonatal mouse NEC model was induced using high-osmolarity formula combined with hypoxia-cold stress. The study observed increased oxidative stress and inflammatory cytokines, alongside activation of ferroptosis markers and intestinal epithelial injury.

Q: What effects did ALAS2 overexpression have on intestinal epithelial cells?

In vitro, ALAS2 overexpression reversed TNF-α/IFN-γ-induced ALAS2 suppression and reduced cell necrosis by 22%. It suppressed ferroptosis markers, decreased lipid reactive oxygen species and mitochondrial depolarization, and improved oxidative damage indicators by restoring antioxidant systems.

Sep 30, 2026

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Neonatal necrotizing enterocolitis (NEC) is a severe gastrointestinal emergency in premature infants, driven by intestinal inflammation, oxidative stress, and progressive regulated cell death. This study evaluates 5′-aminolevulinate synthase 2 (ALAS2) in NEC-induced intestinal injury using neonatal mouse models and intestinal epithelial cell experiments. NEC induction downregulates ALAS2 while increasing oxidative stress, inflammatory cytokines, and ferroptosis activation. Lentivirus-mediated ALAS2 overexpression reverses necrosis and ferroptosis markers, reduces oxidative damage, and reshapes nutrient-redox metabolic pathways, identifying ALAS2 as a potential therapeutic target.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages15

Reading Time38 min

AccessFree to download

Introduction

NEC epidemiology and clinical impact

Inflammation and oxidative stress in NEC

Regulated cell death and ferroptosis

Role of iron metabolism and ALAS2

Materials and methods

Results

ALAS2 downregulation and NEC-associated changes

ALAS2 overexpression blocks necrosis and ferroptosis

Oxidative damage and redox restoration

Untargeted metabolomics findings

Discussion

ALAS2-mediated mechanism and therapeutic implications

Conclusions

Wiley Mediators of Inflammation Volume 2026, Article ID 6683001, 15 pages [https://doi.org/10.1155/mi/6683001](https://doi.org/10.1155/mi/6683001) Research Article ALAS2 Prevents Neonatal Necrotizing Enterocolitis by Improving Ferroptosis in Intestinal Epithelial Cells Through Inhibition of Oxidative Stress Zenghui Hao , 1 Jinbao Han ,2 Ting Yao , 1 Zheng Zhao , 1 Wei Fan , 1 Zaiqun Jiang , 1 Yunting Wang , 1 Xiaoqian Yang , 1 and Zhilin Xu 1 1Department of Pediatric Surgery, The Sixth Afﬁliated Hospital of Harbin Medical University, Harbin, China 2Department of Emergency Center, Beijing Children ’s Hospital, Capital Medical University, National Center for Children ’s Health, Beijing, China Correspondence should be addressed to Zhilin Xu; [xzlin333@163.com](xzlin333@163.com) Received 15 January 2025; Revised 21 August 2025; Accepted 14 November 2025 Academic Editor: Jasbir Arora (RSS) Copyright © 2026 Zenghui Hao et al. Mediators of Inﬂammation published by John Wiley & Sons Ltd. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. Neonatal necrotizing enterocolitis (NEC) is an intestinal disease that occurs in the neonatal period. The purpose of this study was to investigate the role of 5′-aminolevulinate synthase 2 (ALAS2) in NEC-induced intestinal injury. In a neonatal mouse, NEC model was induced by high-osmolarity formula and hypoxia-cold stress, and ALAS2 expression was signiﬁcantly downregulated inileal tissues (p < 0:01), coinciding with elevated oxidative stress (increased Fe2+/malondialdehyde [MDA] and decreased superoxide dismutase [SOD]), inﬂammation (increased TNF-α/interferon-gamma [IFN-γ]), and ferroptosis activation (increased acylCoA synthetase long-chain family member 4 [ACSL4] and decreased ferritin heavy chain 1 [FTH1] with mitochondrial shrinkage) . In vitro, tumor necrosis factor-alpha (TNF-α)/IFN-γ-treated intestinal epithelial cell (IEC) exhibited progressive ALAS2 suppression and increased necrosis. Crucially, lentivirus-mediated ALAS2 overexpression reversed these effects, reducing cell necrosis by 22% while suppressing ferroptosis markers (Fe2+ accumulation, lipid reactive oxygen species [ROS], and mitochondrial depolarization) and oxidative damage (decreased MDA and restored glutathione [GSH]/catalase [CAT]/SOD) . Untargeted metabolomics further revealed ALAS2-mediated modulation of nutrient metabolism and redox pathways. Collectively, ALAS2 ameliorates NEC by blocking oxidative stress-driven ferroptosis in IECs, proposing a novel therapeutic target. Keywords: ALAS2; ferroptosis; neonatal necrotizing enterocolitis; oxidative stress 1. Introduction Neonatal necrotizing enterocolitis (NEC) remains a devastating gastrointestinal emergency predominantly affecting premature infants, with an incidence ranging from 5% to 10% in very low birth weight infants and associated mortality rates reaching 20%–30%[1]. Despite advances in neonatal care, NEC continues to be a leading cause of morbidity and mortality in neonatal intensive care units worldwide [2]. This disease is characterized by intestinal inﬂammation, barrier disruption, and, ultimately, necrosis of the bowel wall [3, 4]. The pathogenesis of NEC is multifactorial and incompletely understood, involving an intricate interplay between prematurity, intestinal dysbiosis, ischemia–reperfusion injury, and a dysregulated host immune response within the immature gut [5, 6] . A critical consequence of these insults is the excessive activation of inﬂammatory cascades and the generation of oxidative stress, which are central drivers of intestinal epithelial cell (IEC) damage and the progression to tissue necrosis [7]. The in-depth exploration of the pathogenesis of NEC could help to further improve the diagnosis and treatment of the disease. Inﬂammation is a hallmark of NEC pathop

FAQs

What role does ALAS2 play in neonatal necrotizing enterocolitis (NEC)?

ALAS2 expression is significantly downregulated in NEC models and is linked to oxidative stress and ferroptosis activation. Restoring ALAS2 via lentiviral overexpression mitigates injury by blocking oxidative stress-driven ferroptosis in intestinal epithelial cells.

How was the NEC model induced and what changes were observed?

A neonatal mouse NEC model was induced using high-osmolarity formula combined with hypoxia-cold stress. The study observed increased oxidative stress and inflammatory cytokines, alongside activation of ferroptosis markers and intestinal epithelial injury.

What effects did ALAS2 overexpression have on intestinal epithelial cells?

In vitro, ALAS2 overexpression reversed TNF-α/IFN-γ-induced ALAS2 suppression and reduced cell necrosis by 22%. It suppressed ferroptosis markers, decreased lipid reactive oxygen species and mitochondrial depolarization, and improved oxidative damage indicators by restoring antioxidant systems.

Sep 30, 2026

Uploaded by Aurelia