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The Role of GPX Enzymes, Lipid Profiles, and Iron Accumulation in Necrotizing Enterocolitis
Grant H Gershner1, Chase Calkins1, Alena Golubkova1
1Division of Pediatric Surgery, Department of Surgery, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA.
Insights
Necrotizing enterocolitis (NEC) involves intestinal inflammation and necrosis in premature infants. This study reveals that oxidative stress, iron dysregulation, and altered lipid metabolism are key contributors to NEC pathogenesis.
Area of Science:
- Gastroenterology
- Neonatal Medicine
- Biochemistry
Background:
- Necrotizing enterocolitis (NEC) is a critical gastrointestinal disease in premature infants.
- Oxidative stress and ferroptosis are implicated in NEC pathogenesis.
- Previous work indicated Glutathione Peroxidase 4 (GPX4) deficiency and lipid radical accumulation in NEC.
Purpose of the Study:
- To investigate the roles of oxidative stress, iron, and lipid metabolism in NEC.
- To analyze human intestinal tissue and NEC-induced human enteroids.
Main Methods:
- Analysis of human intestinal tissue (RNA, protein, immunohistochemistry, immunofluorescence, ICP-MS for iron/selenium).
- Induction of NEC in human enteroids (LPS and hypoxia).
- Lipidomic analysis, NADPH quantification, and iron assays (bathophenanthroline, ICP-MS).
Main Results:
- Patients with NEC showed significantly higher GPX2 levels (p=0.0003) and iron levels (p=0.0102).
- NEC-induced enteroids had decreased NADPH compared to initial controls (p=0.0091).
- Significant alterations in lipid distribution were observed in NEC patients, but not in fatty acid profiles.
Conclusions:
- Oxidative stress, iron dysregulation, and altered lipid metabolism are significant contributors to NEC pathogenesis.
- GPX2 upregulation and iron accumulation are associated with NEC.
- Further research into these pathways may yield novel therapeutic strategies for NEC.
Abstract:
Necrotizing enterocolitis (NEC) is a serious GI disease of premature infants, marked by intestinal inflammation and necrosis. Recent research has highlighted the potential role of oxidative stress (OS) and ferroptosis in its pathogenesis. We previously identified a deficiency in Glutathione Peroxidase (GPX) 4 and lipid radical accumulation, prompting further investigation. Human intestinal tissue from a prior study was processed, and it underwent RNA and protein isolation, Immunohistochemistry, Immunofluorescence, and acid digestion for iron and selenium analysis via Inductively coupled mass spectrometry (ICP-MS). NEC was induced in human enteroids using lipopolysaccharide (LPS) and hypoxia, followed by RNA/protein isolation and lipidomic analysis. Humans with NEC had significantly higher levels of GPX2 (p = 0.0003). Enteroids exposed to NEC conditions had significantly decreased amounts of NADPH compared to initial controls (p = 0.0091), but similar levels compared to post-24 h controls (p = 0.3520). Patients with NEC had significantly higher levels of iron compared to controls via the bathophenanthroline-based assay (p = 0.0102) and with ICP-MS (p = 0.0148). There were several significant alterations in lipid distribution between NEC and control patients, but not in the fatty acid profiles. Our study suggests that oxidative stress, iron dysregulation, and altered lipid metabolism contribute to NEC pathogenesis.
