Effects of Tris and histidine on human erythrocytes and conditions influencing their mode of action.

1. The incubation of human erythrocytes in 0.172 M Tris - HCl, pH 7.6 buffer at 37 degrees leads to (1) a pronounced cellular volume increase, (2) a preferential release of Na+, and (3) if continued sufficiently long, hemolysis. These effects are pH dependent and also are influenced to a considerable degree by such diverse reagents as NaC glucose, and histidine. In each instance, increasing levels of the latter compounds in a Tris - HCl incubation mixture led to diminished cellular volume increase and prolonged time of onset of hemolysis. 2. Histidine solutions of 0.31 M, pH 7.5 caused a rapid and dramatic decrease in cellular volume of human erythrocytes and a concomitant rapid exit of cations. However, in a prolonged incubation, human erythrocytes slowly regained their cell volume as a result of histidine entry into the cell. Of considerable interest: Tris swollen cells undergo immediate shrinkage to far below the initial cell volume when incubated in histidine at 37 degrees C. Through repetition of this process two additional times, as much as 90-95% of the total cellular Na+ and K+ was removed without hemolysis. 3. Human erythrocytes washed in 0.12 M MgCl2 and then suspended in 0.31 M histidine, pH 7.5, lost upwards of 60% of their total Na+ and 30% of their total K+ after a 40 min incubation at 37 degrees C. However, when increasing amounts of 0.172 M Tris - HCl, pH 7.6 were added to the histidine suspension of cells, the release of K+ was reduced to 5% but the release of Na+ decreased only to 40% of the total cellular level. On the basis of these observations, it is evident that Tris exerts a preferential activity towards the efflux of Na+ from the human erythrocyte, whereas histidine results in high efflux of K+ and Na+ from the cell.