Cellular redox potential and hemoglobin S-glutathionylation in human and rat erythrocytes: A comparative study

The rat is commonly used to evaluate responses of red blood cells (RBCs) to oxidative stress. How closely the rat RBC model predicts the human RBC human response has not been well characterized. The objective of this study was to compare human and rat RBC responses to the thiol-specific oxidant tert-butylhydroperoxide by monitoring the intraerythrocyte glutathione redox potential and its correlation with hemoglobin S-glutathionylation. Changes in redox potential did not differ significantly between rat and human RBCs under the considered conditions, and both human and rat hemoglobins were apparently S-glutathionylated by a thiol-disulfide exchange mechanism with glutathione disulfide, though the extent of S-glutathionylation in rat erythrocytes was more than 10-fold higher than in human ones. On the contrary, human and rat hemoglobin S-glutathionylation differently correlated with redox potential for the glutathione redox couple, suggesting that the formation of S-glutathionylated hemoglobin was not simply a function of glutathione disulfide concentration or glutathione/glutathione disulfide ratio and that the content of reactive cysteines in hemoglobin beta globin can strongly influence intraerythrocyte glutathione metabolism and distribution between free and hemoglobin-bound forms. This study reveals fundamental physiological differences in rat and human RBCs because of differences in rat and human beta globin cysteine and reactivity, which can have important implications for the study of rat biology as a whole and for the use of rats as models for human beings under physiological and pathological circumstances and, therefore, highlights the need for caution when extrapolating rat responses to humans.