Andrea Ferrigno1, Laura Giuseppina Di Pasqua1, Alberto Bianchi1, Plinio Richelmi1, Mariapia Vairetti1. 1. Andrea Ferrigno, Laura Giuseppina Di Pasqua, Alberto Bianchi, Plinio Richelmi, Mariapia Vairetti, Unit of Cellular and Molecular Pharmacology and Toxicology, Department of Internal Medicine and Therapeutics, University of Pavia, Pavia 27100, Italy.
Abstract
AIM: To study at what temperature the oxygen carried by the perfusate meets liver requirements in a model of organ perfusion. METHODS: In this study, we correlated hypoxia inducible factor (HIF)-1α expression to the perfusion temperature and the hepatic oxygen uptake in a model of isolated perfused rat liver. Livers from Wistar rats were perfused for 6 h with an oxygenated medium at 10, 20, 30 and 37 °C. Oxygen uptake was measured by an oxygen probe; lactate dehydrogenase activity, lactate release and glycogen were measured spectrophotometrically; bile flow was gravitationally determined; pH of the perfusate was also evaluated; HIF-1α mRNA and protein expression were analyzed by real time-polymerase chain reaction and ELISA, respectively. RESULTS: Livers perfused at 10 and 20 °C showed no difference in lactate dehydrogenase release after 6 h of perfusion (0.96±0.23 vs 0.93±0.09 mU/min per g) and had lower hepatic damage as compared to 30 and 37 °C (5.63±0.76 vs 527.69±45.27 mU/min per g, respectively, Ps<0.01). After 6 h, tissue ATP was significantly higher in livers perfused at 10 and 20 °C than in livers perfused at 30 and 37 °C (0.89±0.06 and 1.16±0.05 vs 0.57±0.09 and 0.33±0.08 nmol/mg, respectively, Ps<0.01). No sign of hypoxia was observed at 10 and 20 °C, as highlighted by low lactate release respect to livers perfused at 30 and 37 °C (121.4±12.6 and 146.3±7.3 vs 281.8±45.3 and 1094.5±71.7 nmol/mL, respectively, Ps<0.02), and low relative HIF-1α mRNA (0.40±0.08 and 0.20±0.03 vs 0.60±0.20 and 1.47±0.30, respectively, Ps<0.05) and protein (3.72±0.16 and 3.65±0.06 vs 4.43±0.41 and 6.44±0.82, respectively, Ps<0.05) expression. CONCLUSION: Livers perfused at 10 and 20 °C show no sign of liver injury or anaerobiosis, in contrast to livers perfused at 30 and 37 °C.
AIM: To study at what temperature the oxygen carried by the perfusate meets liver requirements in a model of organ perfusion. METHODS: In this study, we correlated hypoxia inducible factor (HIF)-1α expression to the perfusion temperature and the hepatic oxygen uptake in a model of isolated perfused rat liver. Livers from Wistar rats were perfused for 6 h with an oxygenated medium at 10, 20, 30 and 37 °C. Oxygen uptake was measured by an oxygen probe; lactate dehydrogenase activity, lactate release and glycogen were measured spectrophotometrically; bile flow was gravitationally determined; pH of the perfusate was also evaluated; HIF-1α mRNA and protein expression were analyzed by real time-polymerase chain reaction and ELISA, respectively. RESULTS: Livers perfused at 10 and 20 °C showed no difference in lactate dehydrogenase release after 6 h of perfusion (0.96±0.23 vs 0.93±0.09 mU/min per g) and had lower hepatic damage as compared to 30 and 37 °C (5.63±0.76 vs 527.69±45.27 mU/min per g, respectively, Ps<0.01). After 6 h, tissue ATP was significantly higher in livers perfused at 10 and 20 °C than in livers perfused at 30 and 37 °C (0.89±0.06 and 1.16±0.05 vs 0.57±0.09 and 0.33±0.08 nmol/mg, respectively, Ps<0.01). No sign of hypoxia was observed at 10 and 20 °C, as highlighted by low lactate release respect to livers perfused at 30 and 37 °C (121.4±12.6 and 146.3±7.3 vs 281.8±45.3 and 1094.5±71.7 nmol/mL, respectively, Ps<0.02), and low relative HIF-1α mRNA (0.40±0.08 and 0.20±0.03 vs 0.60±0.20 and 1.47±0.30, respectively, Ps<0.05) and protein (3.72±0.16 and 3.65±0.06 vs 4.43±0.41 and 6.44±0.82, respectively, Ps<0.05) expression. CONCLUSION: Livers perfused at 10 and 20 °C show no sign of liver injury or anaerobiosis, in contrast to livers perfused at 30 and 37 °C.
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