Journal
TOXICOLOGY IN VITRO
Volume 21, Issue 5, Pages 835-839Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.tiv.2007.02.002
Keywords
verapamil; haemolysis; equine red blood cell; calcium; osmotic stress
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The haemolytic effect of verapamil on red blood cells (RBCs) exposed to varying osmolarity was investigated. The experimental approach used a modified red cell haernolysis assay with concentrations of verapamil ranging from 50-1500 M compared to drug free controls. The time-course of haemolytic effects was also investigated. We also briefly determined the haemolytic effects of verapamil in Ca2+-free conditions (with added EGTA). In conditions representing decreasing osmolarity (dilution from 140-0 mM NaCI) there was a significant increase in erythrocyte haemolysis that was also dependent on verapamil concentration (ANOVA, p < 0.05). The red cells also showed a significantly increased rate of haemolysis over 5 h with increasing verapamil concentration (ANOVA, p < 0.05). The degree of RBC hypotonic haemolysis was significantly increased in a Ca2+-free medium (+EGTA) compared to normal saline and this effect was exacerbated by additions of verapamil (ANOVA, p < 0.05). Overall the data suggested that verapamil can cause haemolysis of RBCs in a predictable time- and concentration-dependent manner, and that verapamil increases the fragility of the erythrocytes further during hypotonic osmotic stress and Ca2+ -free conditions. The mechanism of veraparnil-dependent haemolysis could be directly related to the observed biphasic concentration-effect and could consequently involve several ion transport pathways. (c) 2007 Elsevier Ltd. All rights reserved.
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