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Updated: May 7, 2026
Murine Model of Intestinal Ischemia-reperfusion Injury
Published on: May 11, 2016
Inhibition of ERK1/2 worsens intestinal ischemia/reperfusion injury
Kechen Ban1, Zhanglong Peng, Rosemary A Kozar
1Department of Surgery, University of Texas Health Science Center at Houston, Houston, Texas, United States of America.
Background:
The role of extracellular signal-regulated protein kinase (ERK) in intestinal ischemia/reperfusion (I/R) injury has not been well investigated. The aim of the current study was to examine the effect of inhibition of the ERK pathway in an in vitro and in vivo model of intestinal I/R injury.
Methods:
ERK1/2 activity was inhibited using the specific inhibitor, U0126, in intestinal epithelial cells under hypoxia/reoxygenation conditions and in mice subjected to 1 hour of intestinal ischemia followed by 6 hours reperfusion. In vitro, cell proliferation was assessed by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay, apoptosis by DNA fragmentation, and migration using an in vitro model of intestinal wound healing. Cells were also transfected with a p70S6K plasmid and the effects of overexpression similarly analyzed. In vivo, the effects of U0126 on intestinal cell proliferation and apoptosis, intestinal permeability, lung and intestinal neutrophil infiltration and injury, and plasma cytokine levels were measured. Survival was also assessed after U0126. Activity of p70S6 kinase (p70S6K) was measured by Western blot.
Results:
In vitro, inhibition of ERK1/2 by U0126 significantly decreased cell proliferation and migration but enhanced cell apoptosis. Overexpression of p70S6K promoted cell proliferation and decreased cell apoptosis. In vivo, U0126 significantly increased cell apoptosis and decreased cell proliferation in the intestine, increased intestinal permeability, intestinal and lung neutrophil infiltration, and injury, as well as systemic pro-inflammatory cytokines, TNF-α, IL-6 and IL-1β. Mortality was also significantly increased by U0126. Inhibition of ERK1/2 by U0126 also abolished activity of p70S6K both in vitro and in vivo models.
Conclusion:
Pharmacologic inhibition of ERK1/2 by U0126 worsens intestinal IR injury. The detrimental effects are mediated, at least in part, by inhibition of p70S6K, the major effector of mammalian target of rapamycin pathway.
Insights
Inhibiting the extracellular signal-regulated protein kinase (ERK) pathway worsens intestinal ischemia/reperfusion (I/R) injury. This detrimental effect is partly due to blocking p70S6K, a key component of the mammalian target of rapamycin pathway.
Area of Science:
- Gastroenterology
- Cell Biology
- Molecular Medicine
Background:
- The role of extracellular signal-regulated protein kinase (ERK) in intestinal ischemia/reperfusion (I/R) injury remains largely unexplored.
- This study investigates the impact of inhibiting the ERK pathway on intestinal I/R injury using both in vitro and in vivo models.
Purpose of the Study:
- To examine the effect of ERK pathway inhibition on intestinal epithelial cells under hypoxia/reoxygenation.
- To evaluate the consequences of inhibiting ERK1/2 on intestinal I/R injury in a murine model.
Main Methods:
- Utilized U0126, a specific ERK1/2 inhibitor, in intestinal epithelial cells and mice subjected to I/R.
- Assessed cell proliferation (MTT), apoptosis (DNA fragmentation), migration, and p70S6K activity (Western blot).
- Measured in vivo parameters including intestinal permeability, neutrophil infiltration, cytokine levels, and survival.
Main Results:
- Inhibition of ERK1/2 decreased cell proliferation and migration while increasing apoptosis in vitro.
- In vivo, U0126 exacerbated intestinal apoptosis, increased permeability, neutrophil infiltration, and pro-inflammatory cytokines (TNF-α, IL-6, IL-1β), leading to higher mortality.
- ERK1/2 inhibition abolished p70S6K activity in both models.
Conclusions:
- Pharmacological inhibition of ERK1/2 exacerbates intestinal I/R injury.
- The detrimental effects are mediated, in part, by the inhibition of p70S6K, a critical effector of the mammalian target of rapamycin pathway.
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