Low pH-induced Hemolysis of Erythrocytes Is Related to the Entry of the Acid Into Cytosole and Oxidative Stress on Cellular Membranes

Biochim Biophys Acta. 1999 Jan 8;1415(2):349-60. doi: 10.1016/s0005-2736(98)00202-8.

Abstract

HCl-induced lysis of mammalian erythrocytes, pretreated with DIDS, which is a specific inhibitor of the anion transport in their membranes, was markedly delayed. After acidification of a suspension of DIDS-inhibited cells, hemolysis was initiated by addition of a protonophore (Na-salicylate) at any moment chosen by will. These findings revealed that low-pH hemolysis depended on the rate of the transfer of acid equivalents into cytosole. Erythrocyte acid resistance was studied in a group of mammals and found to be inversely related to the rate of monovalent anion exchange in membranes which supported the above observations. In human erythrocytes, the critical level of cytosole acidification was found to be about pH 5.7 by measuring the acid equivalent absorbed by cells prior to hemolysis. HCl-induced hemolysis was also studied in human erythrocyte ghosts resealed with one-sixth of the initial hemoglobin content of cells. During the prelytic interval the ghosts suspended in isotonic NaCl/sucrose media shrunk, indicating an increase in ion permeability. The increase in prelytic permeability and hemolysis were strongly delayed in ghosts prepared from DIDS-treated cells, suggesting a uniform mechanism of lysing in cells and their ghosts. The prelytic increase in ion permeability was measured by the corresponding rate of ghost shrinkage and was found to be pH-dependent, with a high value below pH 3.4 and a very low one above pH 4.0. Compared to cells, the prelytic barrier impairment in ghosts had more mild character although it required greater concentration of cytosolic H+. While finally complete, hemolysis of cells was strongly delayed in the presence of catalase (500-1500 U/ml) and superoxide dismutase (200-600 U/ml) in hemolytic media. In conclusion, the acid-induced hemolysis could be associated with an oxidative injury of membranes, mainly triggered by the entry of acid equivalents into the cytosole.

MeSH terms

  • 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid
  • Animals
  • Cattle
  • Cell Membrane Permeability
  • Cytosol / metabolism
  • Erythrocyte Membrane / immunology
  • Erythrocyte Membrane / metabolism*
  • Erythrocytes / immunology*
  • Hemolysis*
  • Humans
  • Hydrogen-Ion Concentration
  • Oxidative Stress
  • Time Factors

Substances

  • 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid