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  • Title: Protection by ascorbate against apoptosis of thymocytes: implications of ascorbate-induced nonlethal oxidative stress and poly(ADP-ribosyl)ation.
    Author: Maellaro E, Bello BD, Comporti M.
    Journal: Exp Cell Res; 1996 Jul 10; 226(1):105-13. PubMed ID: 8660945.
    Abstract:
    Apoptosis can be triggered in thymocytes with stimuli (6alpha-methylprednisolone, thapsigargin, and etoposide) acting by different mechanisms. In each of these instances cell death is extensively prevented until 5 h of incubation when cells are preincubated with 250 microM ascorbic acid (AA) for 1 h, then washed, and incubated in fresh medium containing the above mentioned apoptotic stimuli. In addition, the degree of spontaneous apoptosis of untreated thymocytes is somewhat lower in the AA-preincubated cells. The protection against apoptosis does not seem to be dependent on the intracellular enrichment of AA, as measured at the end of the preincubation period. On the contrary, such a protection is strictly related to a partial loss of ascorbate in the medium (possibly due to its autooxidation), is catalase-inhibitable, and is reproduced by a preincubation of the cells with nontoxic concentrations of hydrogen peroxide. The AA-supplemented cells show a remarkable decrease in NAD+ levels and a significant increase of poly(ADP-ribose) polymerase (PARP) activity. Consistently with these results, the addition of PARP inhibitors, such as thymidine and 3-aminobenzamide, during the preincubation with AA, prevents NAD+ depletion and abolishes the protective effect of AA against apoptosis. The possibility is discussed that an early activation of PARP by stimuli which are nontoxic per se makes the cells able to withstand subsequent apoptotic stimuli which are otherwise lethal.
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