Calcium potentiates the peroxidation of erythrocyte membrane lipids

To explore the possible role of intracellular calcium in membrane lipid peroxidation, we subjected red cells to conditions designed to increase intracellular calcium levels and then measured lipid peroxidation after exposure to a peroxidant threat. Human erythrocytes were pretreated for 3 h with either very high levels of CaCl2, or with low levels in the presence of the ionophore A23187. The erythrocytes were subsequently exposed to a peroxide-generating system consisting of xanthine and xanthine oxidase, or H2O2 for 1 h at 37 degrees C. As measured by a malonyldialdehyde assay, the calcium-treated cell showed up to a 2-fold increase in lipid peroxidation in comparison to untreated cells. In experiments with the ionophore, calcium concentration-dependent effects were detected at levels as low as 10 microM and were maximal at 50 microM. A significant loss of phosphatidylserine and phosphatidylethanolamine was observed in calcium- and peroxide-treated erythrocytes. This potentiation of membrane lipid peroxidation and lipid loss could be prevented by either lipid antioxidants or EGTA. The present study shows that pretreatment of erythrocytes with calcium increases their sensitivity to lipid peroxidation. This suggests that increased calcium concentration may be a factor in the potentiation of membrane lipid peroxidation of erythrocytes known to have increased calcium levels such as sickled and senescent red cells.