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Updated: Jun 23, 2026
Murine Model of Intestinal Ischemia-reperfusion Injury
Published on: May 11, 2016
Intestinal ischemia-reperfusion injury: reversible and irreversible damage imaged in vivo
Yanfang Guan1, Roger T Worrell, Timothy A Pritts
1Department of Molecular and Cellular Physiology, University of Cincinnati College of Medicine, 231 Albert Sabin Way, Cincinnati, OH 45267-0576, USA. guany@uc.edu
Abstract:
The early events in an intestinal ischemic episode have been difficult to evaluate. Using in vivo microscopy we have analyzed in real-time the effects of short (15 min) and long (40-50 min) ischemia with subsequent reperfusion (IR), evaluating structure, integrity, and functioning of the mouse jejunal mucosa while monitoring blood flow by confocal microscopy. IR was imposed by inflation/deflation of a vascular occluder, and blood flow was monitored and confirmed with scanning confocal imaging. After short ischemia, villus tip cells revealed a rapid increase (23%) in the intracellular NAD(P)H concentration (confocal autofluorescence microscopy), and the pH-sensitive probe BCECF showed a biphasic response of the intracellular pH (pH(i)), quickly alkalinizing from the resting value of 6.8 +/- 0.1 to 7.1 +/- 0.1 but then strongly acidifying to 6.3 +/- 0.1. Upon reperfusion, values returned toward control. In contrast, results were heterogeneous after long IR. During long ischemia, one-third of the epithelial cells remained viable with reversible changes upon reperfusion, but remaining cells lost membrane integrity (Lucifer Yellow uptake, LY) and had membrane blebs during ischemia. These effects became more pronounced as the reperfusion interval progressed when cells exhibited more severely affected NAD(P)H and pH(i) values, larger blebs, and more LY uptake and eventually were shed from the villus. Results from stereo microscopy suggest that these irreversible effects of IR may have occurred as a result of incomplete restorations of local blood flow, especially at the antimesenteric side of the intestine. We conclude that the adverse effects of short ischemia on the jejunum epithelium are fully reversible during the reperfusion interval. However, after long ischemia, reperfusion cannot restore normal structure and functioning of a majority of cells, which deteriorate further. Our results provide a basis for defining the cellular events that cause tissue to transit from reversible to irreversible damage during IR.
Insights
Short intestinal ischemia (15 min) causes reversible damage to jejunal mucosa. Longer ischemia (40-50 min) leads to irreversible cell damage and loss, even after reperfusion, highlighting critical time windows for intervention.
Area of Science:
- Gastroenterology
- Cell Biology
- Physiology
Background:
- Evaluating early events in intestinal ischemia is challenging.
- Understanding the impact of ischemia-reperfusion (IR) on mucosal integrity is crucial.
Purpose of the Study:
- To analyze the real-time effects of short and long ischemia with reperfusion on mouse jejunal mucosa structure, integrity, and function.
- To monitor blood flow changes during IR using confocal microscopy.
Main Methods:
- In vivo microscopy and scanning confocal imaging to assess jejunal mucosa and blood flow.
- Confocal autofluorescence microscopy to measure intracellular NAD(P)H concentrations.
- BCECF probe to evaluate intracellular pH (pH(i)) and Lucifer Yellow uptake for membrane integrity.
Main Results:
- Short ischemia (15 min) caused transient NAD(P)H increase and biphasic pH(i) changes, fully reversible upon reperfusion.
- Long ischemia (40-50 min) resulted in heterogeneous outcomes: one-third of cells recovered, while others lost membrane integrity, showed blebbing, and deteriorated further post-reperfusion.
- Stereo microscopy suggested incomplete blood flow restoration contributed to irreversible damage.
Conclusions:
- Short-term intestinal ischemia is fully reversible in the jejunum.
- Prolonged ischemia leads to irreversible mucosal damage, with reperfusion unable to restore normal cell structure and function in a majority of cells.
- These findings define cellular events differentiating reversible from irreversible IR injury.
