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Updated: Oct 24, 2025
Renal Ischaemia Reperfusion Injury: A Mouse Model of Injury and Regeneration
Published on: June 7, 2014
Differential Regulation of Damage-Associated Molecular Pattern Release in a Mouse Model of Skeletal Muscle
Hiroaki Furubeppu1, Takashi Ito2, Midori Kakuuchi1
1Department of Emergency and Intensive Care Medicine, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, Japan.
Background:
Skeletal muscle ischemia/reperfusion (I/R) injury is an important clinical issue that can cause remote organ injury. Although its pathogenesis has not been fully elucidated, recent studies have suggested that damage-associated molecular patterns (DAMPs) are mediators of remote organ injury in sterile inflammation. The purpose of this study was to investigate the possible involvement of DAMPs, including the nuclear proteins high-mobility group box 1 (HMGB1) and histone H3, in the pathogenesis of skeletal muscle I/R injury in mice.
Methods:
Hindlimb ischemia was induced in mice through bilateral ligation of inguinal regions using rubber grommets. Reperfusion was induced by cutting the rubber grommets after 2-12 h of ischemic period. Survival rates, localization of HMGB1 and histone H3 in the gastrocnemius muscle, and circulating HMGB1 and histone H3 levels were analyzed. The effect of anti-HMGB1 and anti-histone H3 antibodies on survival was analyzed in mice with I/R injury.
Results:
All mice with hindlimb ischemia survived for at least 36 h, while all mice died within 24 h if the hindlimbs were reperfused after ischemia for 4-12 h. Immunohistochemical analysis revealed that HMGB1 translocated from the nucleus to the cytoplasm in the ischemic gastrocnemius muscle, while histone H3 was confined to the nucleus. Accordingly, serum HMGB1 levels were significantly elevated in mice with hindlimb I/R compared with normal mice or mice with hindlimb ischemia (P < 0.05). Serum histone H3 levels were not elevated after I/R. Treatment with anti-HMGB1 antibodies significantly improved survival of mice with hindlimb I/R injury compared with control antibodies (P < 0.05).
Conclusions:
HMGB1, but not histone H3, translocated to the cytoplasm during skeletal muscle ischemia, and was released into the systemic circulation after reperfusion in mice with I/R injury. Treatment with anti-HMGB1 antibodies partially improved survival.
Insights
High-mobility group box 1 (HMGB1) is released during skeletal muscle ischemia/reperfusion (I/R) injury, contributing to mortality. Neutralizing HMGB1 with antibodies improved survival rates in mice, highlighting its role in I/R pathogenesis.
Area of Science:
- Cellular and Molecular Biology
- Immunology
- Physiology
Background:
- Skeletal muscle ischemia/reperfusion (I/R) injury is a significant clinical concern with potential for remote organ damage.
- Damage-associated molecular patterns (DAMPs) are implicated in sterile inflammation and remote organ injury following I/R.
- High-mobility group box 1 (HMGB1) and histone H3 are nuclear proteins investigated as potential DAMPs in I/R injury.
Purpose of the Study:
- To investigate the role of HMGB1 and histone H3 in the pathogenesis of skeletal muscle I/R injury in a mouse model.
- To determine the cellular localization and systemic release of HMGB1 and histone H3 following I/R.
- To evaluate the therapeutic potential of targeting HMGB1 and histone H3 in I/R injury.
Main Methods:
- Induction of hindlimb ischemia and subsequent reperfusion in mice.
- Analysis of survival rates, HMGB1 and histone H3 localization in gastrocnemius muscle via immunohistochemistry.
- Measurement of circulating HMGB1 and histone H3 levels.
- Assessment of the effect of anti-HMGB1 and anti-histone H3 antibodies on survival.
Main Results:
- Hindlimb I/R injury led to significant mortality within 24 hours.
- HMGB1 translocated from the nucleus to the cytoplasm in ischemic muscle, and serum HMGB1 levels were significantly elevated post-reperfusion.
- Histone H3 remained nuclear, and serum levels did not increase.
- Anti-HMGB1 antibody treatment significantly improved survival rates compared to control antibodies.
Conclusions:
- HMGB1, but not histone H3, undergoes cytoplasmic translocation and systemic release during skeletal muscle I/R.
- HMGB1 is a key mediator in the pathogenesis of skeletal muscle I/R injury.
- Targeting HMGB1 with antibodies offers a potential therapeutic strategy to improve outcomes in I/R injury.
