Ferrigno Andrea, Di Pasqua Laura Giuseppina, Bianchi Alberto, Richelmi Plinio, Vairetti Mariapia
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.
World J Gastroenterol. 2015 Jan 28;21(4):1108-16. doi: 10.3748/wjg.v21.i4.1108.
To study at what temperature the oxygen carried by the perfusate meets liver requirements in a model of organ perfusion.
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.
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.
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.
在器官灌注模型中研究灌注液携带的氧气在何种温度下能满足肝脏需求。
在本研究中，我们在离体灌注大鼠肝脏模型中，将缺氧诱导因子（HIF）-1α表达与灌注温度及肝脏氧摄取相关联。用含氧培养基在10、20、30和37°C下对Wistar大鼠的肝脏进行6小时灌注。通过氧探针测量氧摄取；用分光光度法测量乳酸脱氢酶活性、乳酸释放和糖原；通过重力法测定胆汁流量；还评估灌注液的pH值；分别通过实时聚合酶链反应和酶联免疫吸附测定法分析HIF-1α mRNA和蛋白表达。
在10和20°C下灌注的肝脏在灌注6小时后乳酸脱氢酶释放无差异（分别为0.96±0.23与0.93±0.09 mU/分钟每克），与30和37°C相比肝脏损伤更低（分别为5.63±0.76与527.69±45.27 mU/分钟每克，P<0.01）。6小时后，在10和20°C下灌注的肝脏中组织ATP显著高于在30和37°C下灌注的肝脏（分别为0.89±0.06和1.16±0.05与0.57±0.09和0.33±0.08 nmol/毫克，P<0.01）。在10和20°C未观察到缺氧迹象，与在30和37°C下灌注的肝脏相比，乳酸释放较低（分别为121.4±12.6和146.3±7.3与281.8±45.3和1094.5±71.7 nmol/毫升，P<0.02），相对HIF-1α mRNA（分别为0.40±0.08和0.20±0.03与0.60±0.20和1.47±0.30，P<0.05）和蛋白表达（分别为3.72±0.16和3.65±0.06与4.43±0.41和6.44±0.82，P<0.05）也较低。
与在30和37°C下灌注的肝脏相比，在10和20°C下灌注的肝脏未显示肝损伤或无氧代谢迹象。
