Purpose The present study was designed to determine whether heavy water containing solution (Dsol) for cold preservation and hydrogen gas for reperfusion conferred protection in rat liver subjected to 48-hour simple cold preservation and subsequent reperfusion. Methods Rat livers were subjected to 48-hour of cold preservation in UW or Dsol solutions, and respective preservation plus hydrogen gas therapy at reperfusion (UW+H2 and Dsol+H2 groups), respectively (n=6). Dsol is a modified UW solution with 30%-D2O. In control (CT) group, grafts were reperfused without preservation. The grafts were reperfused by an isolated perfused rat liver apparatus for 90 min. Portal resistance, bile production, oxygen consumption rate, and perfusate LDH were assessed. Liver sample at the end of reperfusion were applied to HE-staining, immunostainings of endothelin-1 and 8-OHdG, and cytoskeletal interity were assessed by western blot. Results In UW group, the highest portal resistance (Fig.1 Rt) with the least oxygen consumption and bile production throughout reperfusion were shown (Fig.1 Lt), resulting in severe tissue injury evidenced by HE-staining, the highest LDH leakage (UW;137±56, CT;21±18 U/L), diffused 8-OHdG staining and sinusoidal endothelin-1 staining. Cytoskeletal derangement was shown by cleaved Fodrin and low levels of phospho-Slingshot and phospho-LIMK-1. All these changes were significantly reduced in UW+H2 and Dsol groups; LDH leakage (85±35 and 58±80 U/L, respectively), and further improved in Dsol+H2 group; LDH leakage (49±30 U/L). Conclusions Distinct treatment against cold preservation and against reperfusion, by heavy water containing solution and hydrogen gas, respectively, conferred protection in rat liver subjected to extended cold preservation and reperfusion.

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