A Novel Multidrug Combination Mitigates Rat Liver Steatosis Through Activating AMPK Pathway During Normothermic Machine Perfusion

BACKGROUND Hepatic steatosis is now the leading cause of liver discards in deceased donors. Previous studies (Y defatting) have successfully reduced the fat content by treating rat steatotic livers on extracorporeal normothermic machine perfusion (NMP) with a multidrug combination including the GW compounds that were linked to an increased risk of carcinogenesis. METHODS We developed a novel multidrug combination by replacing the GW compounds with 2 polyphenols, epigallocatechin-3-gallate (E) and resveratrol (R). Sixteen rat livers were placed on NMP and assigned to control, Y defatting, Y+E+R defatting, or Y'-GW+E+R defatting groups (Y'-GW=90% dose-reduced Y defatting, n=4/group). RESULTS All livers in defatting groups had significant decreases in hepatic TG content at the end of the experiment. However, livers treated with our novel Y'-GW+E+R combination had evidence of increased metabolism, and less hepatocyte damage and carcinogenic potential. Our Y'-GW+E+R combination had increased phosphorylation of AMPK (P=0.019) and acetyl-coA carboxylase (P=0.023) compared with control; these were not increased in Y+E+R, and actually decreased in the Y groups. Furthermore, the Y'-GW+E+R group had less evidence of carcinogenic potential with no increase in AKT phosphorylation compared to control (P=0.089); the Y (P=0.031) and Y+E+R (P=0.035) groups had striking increases in AKT phosphorylation. Finally, our Y'-GW+E+R showed less evidence of hepatocyte damage with significantly lower perfusate ALT (P=0.007) and AST (P=0.014) levels. CONCLUSIONS We have developed a novel multidrug combination demonstrating promising defatting efficacy via activation of the AMPK pathway with an optimized safety profile and reduced hepatotoxicity during ex vivo NMP.Supplemental Visual Abstract; http://links.lww.com/TP/C139.