Heavy metals and lipofuscinogenesis. A study on myocardial cells cultured under varying oxidative stress

The aim of this study was to examine the influence on lipofuscinogenesis of a number of transition and non-transition heavy metals in cultured post-mitotic cells (neonatal rat myocytes) at varying oxidative stress. The effects of Al, Cd, Cr, Cu, Hg, Pb and Zn, added to the medium as chlorides, were examined after 14 days in culture under 5, 20 and 40% ambient oxygen. Lipofuscin was quantified by microspectrofluorometry of individual cells. The addition of Al (40 microM), Cd (40 nM), Hg (30 microM) and Pb (40 microM) to the culture growth medium markedly increased the amount of intracellular lipofuscin, whereas Cr (40 microM), Cu (40 microM) and Zn (40 microM) had the opposite effect. Transmission electron microscopic examination of the myocytes showed greatly increased numbers of autophagic vacuoles in cells exposed to those heavy metals that increased lipofuscin formation. This effect was most pronounced when cells were grown at high (40%) oxygen tension. Possible explanations for the metal augmented pigment formation may be (i) inhibition of lysosomal enzymes, (ii) catalytic interference with peroxidative reactions, or (iii) general toxicity with unspecifically increased autophagocytosis. The decreased pigment accumulation after the addition of Zn, Cr and Cu may, at least partly, be related to the replacement of iron, which has catalytic activity in Fenton reactions.