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Updated: Aug 6, 2026
A Reproducible Intensive Care Unit-Oriented Endotoxin Model in Rats
Published on: February 20, 2021
Endotoxin-induced Ferroptosis-associated Leukocyte Injury and DAMP Release in Sepsis: An in Vitro Study
Background:
Sepsis is characterized by dysregulated inflammation and thromboinflammation in which regulated cell death and damage-associated molecular patterns (DAMPs) contribute to organ injury. Ferroptosis has recently emerged as a potential mechanism of inflammatory cell injury in sepsis.
Objective:
To investigate ferroptosis-associated leukocyte injury and inflammatory lytic cell death in an in vitro endotoxin model.
Methods:
Rat peritoneal leukocytes were stimulated with lipopolysaccharide (LPS; 0.4 mg/mL). Intracellular, mitochondrial, and lysosomal Fe2+ accumulation was evaluated using FerroOrange®, Mito-FerroGreen®, and Lyso-Ferro Red® fluorescence probes. Cellular morphology was assessed using May-Grünwald-Giemsa staining, DAPI staining, and immunofluorescence for histone H3 and citrullinated histone H3.
Results:
LPS stimulation induced marked leukocyte injury characterized by membrane rupture, cytoplasmic collapse, and extracellular dispersion of cellular contents. Fluorescence imaging demonstrated increased intracellular, mitochondrial, and lysosomal Fe2+ accumulation, suggesting ferroptosis-associated iron dysregulation. DAPI staining showed chromatin injury and extracellular DNA release. Histone H3-positive and citrullinated histone H3-positive cells coexisted within the same inflammatory population, indicating overlap between inflammatory lytic cell death and NETosis-like responses.
Conclusions:
Ferroptosis-associated iron accumulation, DAMP release, and overlapping inflammatory cell death phenotypes may collectively contribute to thromboinflammation and organ injury in sepsis.
Insights
Ferroptosis, a cell death pathway involving iron, contributes to sepsis-induced leukocyte injury. This study shows iron dysregulation and cell damage in an endotoxin model, linking ferroptosis to sepsis pathology.
Area of Science:
- Cell Biology
- Immunology
- Pathology
Background:
- Sepsis involves inflammation and thromboinflammation, where cell death and damage-associated molecular patterns (DAMPs) cause organ injury.
- Ferroptosis is an emerging mechanism of inflammatory cell injury in sepsis.
Purpose of the Study:
- Investigate ferroptosis-associated leukocyte injury.
- Examine inflammatory lytic cell death in an in vitro endotoxin model.
Main Methods:
- Rat peritoneal leukocytes stimulated with lipopolysaccharide (LPS).
- Evaluated intracellular iron accumulation using fluorescence probes (FerroOrange®, Mito-FerroGreen®, Lyso-Ferro Red®).
- Assessed cell morphology and histone modifications via staining and immunofluorescence.
Main Results:
- LPS induced leukocyte injury with membrane rupture and content release.
- Demonstrated increased intracellular, mitochondrial, and lysosomal iron, indicating ferroptosis.
- Observed chromatin injury, extracellular DNA release, and overlapping inflammatory cell death with NETosis-like responses.
Conclusions:
- Ferroptosis-associated iron accumulation and DAMP release contribute to sepsis.
- Overlapping inflammatory cell death phenotypes may drive thromboinflammation and organ injury in sepsis.
