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Updated: Jul 10, 2026
Method of Direct Segmental Intra-hepatic Delivery Using a Rat Liver Hilar Clamp Model
Published on: April 2, 2017
Matrix metalloproteinase inhibition protects rat livers from prolonged cold ischemia-warm reperfusion injury
Virginie Defamie1, Marina Laurens, Damiano Patrono
1Institut de Pharmacologie Moléculaire et Cellulaire, CNRS, UMR6097, Université de Nice-Sophia Antipolis, France.
Unlabelled:
Matrix metalloproteinases (MMPs) have been implicated in the hepatic injury induced after cold ischemia-warm reperfusion (CI-WR), by altering the extracellular matrix (ECM), but their precise role remains unknown. The hepatic MMP expression was evaluated after 2 conditions of CI (4 degrees C for 24 and 42 hours: viable and nonviable livers) followed by different periods of WR, using isolated perfused rat livers. CI-WR induced moderate changes in hepatic MMP transcript levels not influenced by CI duration, whereas gelatinase activities accumulated in liver effluents. Therefore, the protective effect of a new phosphinic MMP inhibitor, RXP409, was tested after prolonged CI. RXP409 (10 microM) was added to the University of Wisconsin solution, and livers were preserved for 42 hours (4 degrees C), then reperfused for 1 hour in Krebs solution (37 degrees C), containing 20% erythrocytes. Liver viability parameters were recorded, and the extent of cell necrosis was evaluated on liver biopsies, using trypan blue nuclear uptake. Treatment with RXP409 significantly improved liver function (transaminase release and bile secretion) and liver injury. In particular, the MMP inhibitor significantly modified the extent of cell death from large clusters of necrotic hepatocytes as found in control livers (2%-60% of liver biopsies; mean, 26% +/- 9%) to isolated necrotic hepatocytes as found in treated livers (0.2%-12%; mean, 3% +/- 2%) (P < 0.05).
Conclusion:
These data demonstrate that MMPs, by altering the ECM, play a major role in liver CI-WR injury leading to extensive hepatocyte necrosis and that their inhibition might prove to be a new strategy in improving preservation solutions.
Insights
Matrix metalloproteinases (MMPs) contribute to liver injury after cold ischemia-warm reperfusion (CI-WR). Inhibiting MMPs with RXP409 significantly reduced hepatocyte necrosis and improved liver function, suggesting a new preservation strategy.
Area of Science:
- Hepatology
- Biochemistry
- Transplantation Biology
Background:
- Matrix metalloproteinases (MMPs) are implicated in hepatic injury following cold ischemia-warm reperfusion (CI-WR).
- The precise role of MMPs in CI-WR injury and their impact on the extracellular matrix (ECM) remain incompletely understood.
- Hepatic MMP expression and activity following varying durations of CI-WR require further investigation.
Purpose of the Study:
- To investigate the role of MMPs in liver injury induced by CI-WR.
- To evaluate the therapeutic potential of a novel phosphinic MMP inhibitor, RXP409, in mitigating CI-WR-induced liver damage.
- To assess the impact of MMP inhibition on hepatocyte necrosis and overall liver function post-CI-WR.
Main Methods:
- Isolated perfused rat livers were subjected to cold ischemia (4°C for 24 or 42 hours) followed by warm reperfusion.
- Hepatic MMP transcript levels and gelatinase activities in liver effluents were measured.
- Livers preserved for 42 hours were treated with RXP409 (10 µM) in University of Wisconsin solution, followed by reperfusion and assessment of liver viability, transaminase release, bile secretion, and hepatocyte necrosis via trypan blue uptake.
Main Results:
- CI-WR induced moderate changes in hepatic MMP transcript levels, irrespective of CI duration.
- Gelatinase activities were elevated in liver effluents post-CI-WR.
- RXP409 treatment significantly improved liver function, reduced transaminase release and increased bile secretion.
- MMP inhibition with RXP409 markedly decreased hepatocyte necrosis from 26% +/- 9% in controls to 3% +/- 2% (P < 0.05).
Conclusions:
- MMPs play a significant role in liver CI-WR injury by altering the ECM, leading to extensive hepatocyte necrosis.
- Inhibition of MMPs represents a promising novel strategy for improving organ preservation solutions and reducing transplant-related liver injury.
- Targeting MMPs could enhance the viability of organs subjected to prolonged ischemia-reperfusion injury.
