The effects of copper and copper. o-phenanthroline complex on the intact human erythrocytes

In the presence of albumin, cytotoxic effects of copper on erythrocytes in vitro were reduced. The presence of o-phenanthroline, on the other hand, accelerated deleterious effects of copper on the erythrocytes resulting in a decrease in GSH level, accumulation of copper in the cell, loss of potassium and gain of sodium, cross-linking of membrane proteins mainly involving spectrin and band 3, and echinocyte formation. These changes were not reversible. Prior treatment of the cells with N-ethylmaleimide inhibited the cross-linking of membrane proteins and echinocytic transformation by copper o-phenanthroline complex (CuP or diamide to a certain extent. Treatment of the cells with vinblastine, a stomatocytogenic agent, restored biconcave disc shape of CuP-treated erythrocytes, although it did not inhibit the cross-linking of spectrin. Blocking of sulfhydryl groups was not the prerequisite for the copper to be accumulated in the erythrocyte. These results may be helpful for the understanding of the mechanism of hemolysis associated with copper intoxication in vivo.