Abstract
The antioxidant dihydrolipoic acid has been shown to reduce hypoxic and excitotoxic neuronal damage in vitro. In the present study, we tested whether pretreatment with α-lipoic acid, which presumably allows endogenous formation of dihydrolipoic acid, can protect cultured neurons against injury caused by cyanide, glutamate, or iron ions, using the trypan blue exclusion method to determine neuronal damage. One hour of preincubation with dihydrolipoic acid (1 μM), but not with α-lipoic acid, reduced damage of neurons from chick embryo telencephalon caused by 1 mM sodium cyanide or iron ions. α-Lipoic acid (1 μM) reduced cyanide-induced neuronal damage when added 24 h before hypoxia, and pretreatment with α-lipoic acid for >24 h enhanced this neuroprotective effect. Both the R- and the S-enantiomer of α-lipoic acid exerted a similar neuroprotective effect. Pretreatment with α-lipoic acid (1 μM) from the day of plating onward prevented the degeneration of chick embryo telencephalic neurons that had been exposed to Fe2+/Fe3+. α-Lipoic acid (1 μM) added to the culture medium the day of plating also reduced neuronal injury induced by 1 mM l-glutamate in rat hippocampal cultures, whereas 30 min of preincubation with α-lipoic acid failed to attenuate glutamate-induced neuronal damage. Our results indicate that neuroprotection by prolonged pretreatment with α-lipoic acid is probably due to the radical scavenger properties of endogenously formed dihydrolipoic acid.