Abstract
High mobility group box 1 (HMGB1) is a nuclear transcription factor. Once HMGB1 is released by damaged cells or activated immune cells, it acts as danger molecule and triggers the inflammatory signaling cascade. Currently, evidence is accumulating that posttranslational modifications such as oxidation may modulate the pro-inflammatory potential of danger signals. We hypothesized that oxidation of HMGB1 may reduce its pro-inflammatory potential and could take place during prolonged ischemia and upon reperfusion.Liver grafts were cold preserved for 24 h and flushed with saline in hourly intervals to collect the effluent. Liver grafts, cold-preserved for 6 h, were transplanted into syngeneic recipients to obtain serum and liver samples 24 h after initiation of reperfusion. Addition of the effluent to a macrophage culture induced the synthesis of tumor necrosis factor-alpha (TNF-α) and interleukin (IL)-6. The stimulatory activity of graft effluent was reduced after depletion of HMGB1 via immunoprecipitation. Oxidation of the effluent HMGB1 using H2O2 attenuated its stimulatory activity as well. Liver transplantation of cold preserved grafts caused HMGB1 translocation and release as determined by immunohistochemistry and ELISA-assay, respectively. Using Western blot with non-reducing conditions revealed the presence of oxidized HMGB1 in liver samples obtained after 12 h and in effluent samples after 16 h of cold preservation as well as in liver and serum samples obtained 24 h after reperfusion.These observations confirm that post-translational oxidation of HMGB1 attenuates its pro-inflammatory activity. Oxidation of HMGB1 as induced during prolonged ischemia and by reoxygenation during reperfusion in vivo might also attenuate its pro-inflammatory activity. Our findings also call for future studies to investigate the mechanism of the inhibitory effect of oxidized HMGB1 on the pro-inflammatory potential.
Highlights
Liver transplantation is currently the only efficient treatment for patients suffering from both acute and chronic liver disease
There was no significant increase in mRNA levels or secreted protein levels for tumor necrosis factor-alpha (TNF-a) or IL-6 by macrophages cultured with effluent collected at 0 h or 4 h after cold preservation of livers
We demonstrated that: (1) high mobility group box 1 (HMGB1), released during cold ischemia, induced the expression of inflammatory mediators by cultured macrophages; (2) oxidation of HMGB1 using H2O2 reduced its pro-inflammatory potential both in vitro and in vivo; and (3) oxidation of HMGB1 took place during prolonged cold ischemia and reperfusion in liver transplantation
Summary
Liver transplantation is currently the only efficient treatment for patients suffering from both acute and chronic liver disease. Significant progress in liver transplantation has been made, primary graft dysfunction remains a problem. Injury inflicted on the graft may result from harvesting, ischemia, preservation and reperfusion. Cold ischemia is an independent risk factor for primary dysfunction and delayed graft function [1]. Cold ischemia activates hepatic sinusoidal epithelial cells and Kupffer cells to produce inflammatory cytokines and to express adhesion molecules [2,3]. Activation of the inflammatory response results in hepatic damage upon liver transplantation. Activation of the inflammatory cascades involves the release of danger signals from injured cells and activated immune cells [4]. An array of danger signals has been identified with high mobility group box 1 (HMGB1) being the most widely explored [5,6,7]
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