Biochemical and ultrastructural evaluation of isolated rat liver systems perfused with a hemoglobin-free medium

Abstract

The viability of hemoglobin-free perfused rat liver was examined with respect to several liver functions and to the intactness of subcellular structures under electron microscopic observation. Provided that rat livers were perfused with the oxygenated buffer solution at a flow rate between 3 and 3.5 ml/min per g of liver, all the biochemical parameters measured in the perfused liver system, i.e. the rates of glucose, pyruvate, and lactate production, the rate of oxygen consumption and the tissue contents of adenine nucleotides, were similar to those observed with perfusion systems containing erythrocytes or albumin. The perfused liver showed a sensitive response to norepinephrine, involving a reduction of pyridine nucleotides and enhancements of glucose production and oxygen consumption. On electron microscopic examination, changes in hepatic-structure indicative of hypoxic injury particularly vacuolar degeneration and mitochondrial swelling, were not detected in the liver after 70 min of perfusion; the fact that the fine structure of the hepatocyte was preserved in all parts of the organ confirmed that the supply of oxygen to the perfused liver was sufficient under the conditions employed. From viewpoint of the generally accepted criteria for the viability of perfused liver, therefore, the results confirmed that the perfusion of liver with a hemoglobin- and albumin-free medium is a convenient and reliable tool for biochemical investigation of the reactions occurring in whole liver.