Impaired autophagic clearance after cold preservation of fatty livers correlates with tissue necrosis upon reperfusion and is reversed by hypothermic reconditioning.
Fatty livers are particularly susceptible to mitochondrial alterations after cold preservation. We thus aimed to improve graft integrity by brief hypothermic oxygenation prior to warm reperfusion. Macrovesicular steatosis was induced in rat livers by fasting and subsequent feeding of a fat-free diet enriched with carbohydrates. Fatty livers were retrieved and stored ischemically at 4 degrees C for 20 hours in histidine-tryptophan-ketoglutarate solution. Hypothermic reconditioning (HR) was performed in some livers by insufflation of gaseous oxygen via the caval vein during the last 90 minutes of preservation. Viability was assessed upon isolated reperfusion. HR resulted in a significant (approximately 5-fold) reduction of parenchymal (alanine aminotransferase and lactate dehydrogenase) and mitochondrial (glutamate dehydrogenase) enzyme release. Functional recovery (bile production, oxygen consumption, and tissue levels of adenosine triphosphate) was significantly improved by HR. In untreated grafts, cellular autophagy (cleavage of LC3B and protein expression of beclin-1) was significantly impaired (<50% of baseline) after preservation/reperfusion but was restored to normal values by HR. HR also increased cleavage of caspase 9 (P < 0.5) and caspase 3 enzyme activity (by a factor of 1.5). In contrast, histological signs of tissue necrosis were abundant after reperfusion in untreated livers and largely abrogated in reconditioned livers. In conclusion, HR limits mitochondrial defects and restores basal rates of cellular autophagy. This may represent a rescue mechanism for maintaining cellular homeostasis and tissue survival.
Minor
Thomas
T
Surgical Research Division, University Clinic of Surgery, Bonn, Germany. chirfor@ukb.uni-bonn.de
Stegemann
Judith
J
Hirner
Andreas
A
Koetting
Martina
M
D000818:Animals; D017209:Apoptosis; D001343:Autophagy; D015925:Cryopreservation / Q000379:methods*; D005234:Fatty Liver / Q000378:metabolism* / Q000628:therapy*; D005947:Glucose / Q000494:pharmacology; D006639:Histidine / Q000737:chemistry; D007036:Hypothermia, Induced / Q000379:methods*; D008297:Male; D008353:Mannitol / Q000494:pharmacology; D009336:Necrosis; D019188:Organ Preservation Solutions / Q000494:pharmacology; D011189:Potassium Chloride / Q000494:pharmacology; D011343:Procaine / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D015424:Reperfusion; D016896:Treatment Outcome; D014364:Tryptophan / Q000737:chemistry
C039984:Bretschneider cardioplegic solution; D019188:Organ Preservation Solutions; D008353:Mannitol; D006639:Histidine; D011343:Procaine; D011189:Potassium Chloride; D014364:Tryptophan; D005947:Glucose