Metal chelators reverse the action of hydrogen peroxide in rat luteal cells

In rat luteal cells hydrogen peroxide (H2O2) interferes with the functional coupling of the luteinizing hormone (LH) receptor and blocks cAMP-dependent progesterone production. To test if this action of H2O2 is dependent on the generation of hydroxyl radicals, the effects of metal chelators and hydroxyl radical scavengers were evaluated. The heavy metal chelator o-phenanthroline prevented H2O2 inhibition of LH-sensitive cAMP and progesterone accumulation and depletion of ATP. Tetrakis(2-pyridylmethyl)ethylenediamine (TPEN) partially reversed inhibition of cAMP accumulation by H2O2 and completely prevented H2O2-induced ATP depletion, but had no effect on H2O2 inhibition of progesterone synthesis. Three other heavy metal chelators, deferoxamine, bathocuproinedisulfonic acid (BA) and penicillamine, as well as hydroxyl radical scavengers ethanol, thiourea and N-(2-mercaptopropionyl)glycine (MPG), had no effect on the luteolytic actions of H2O2. Differential effects of the chelators were probably due to differences in their cell permeability and subcellular compartmentalization. We conclude that metal chelators block the luteolytic actions of H2O2 by a mechanism probably linked to inhibition of hydroxyl radical generation.