Endothelial cells as a source of oxygen-free radicals. An ESR study

Abstract

Endothelial cells were subjected to anoxia/reoxygenation in order to simulate some of the free radical mechanisms occurring in ischaemia/reperfusion. With ESR and spin trapping using the spin traps 5,5-dimethyl-1-pyrroline-1-oxide (DMPO) and 3,3,5,5-dimethyl-1-pyrroline-1-oxide (M4PO), the results show that upon reoxygenation of endothelial cells, following a period of anoxia, these cells generate superoxide (O2-.). Cytotoxicity of the spin traps was measured by standard trypan blue exclusion methods. Cell injury or death was measured at various times during reoxygenation by lactate dehydrogenase (LDH) release. Experiments using oxyupurinol, SOD, CAT and a combination of SOD and CAT show that while oxypurinol partially prevents spin adduct formation, the combination of SOD and CAT is more effective in doing so. These results suggest that the majority of the oxygen radicals produced by endothelial cells are done so exogenously. The results also suggest that endothelial cells are not only a source of oxygen radicals but also a target.