Two distinct mechanisms of nitric oxide-mediated neuronal cell death show thiol dependency

To better understand the mechanism(s) underlying nitric oxide (. NO)-mediat ed toxicity, in the presence and absence of concomitant oxidant exposure, p ostmitotic terminally differentiated NT2N cells, which are incapable of pro ducing . NO, were exposed to PAPA-NONOate (PAPA/NO) and 3-morpholinosydnoni mine (SIN-1). Exposure to SIN-1, which generated peroxynitrite in the range of 25-750 nM/min, produced a concentration- and time-dependent delayed cel l death. In contrast, a critical threshold concentration (>440 nM/min) was required for . NO to produce significant cell injury. Examination of cells by electron microscopy shows a largely necrotic injury after peroxynitrite exposure but mainly apoptotic-like morphology after . NO exposure. Cellular levels of reduced thiols correlated with cell death, and pretreatment with N-acetylcysteine (NAC) fully protected from cell death in either PAPA/NO o r SIN-1 exposure. NAC given within the first 3 h posttreatment further dela yed cell death and increased the intracellular thiol level in SIN-1 but not . NO-exposed cells. Cell injury from . NO was independent of cGMP, caspase s, and superoxide or peroxynitrite formation. Overall, exposure of non-. NO -producing cells to . NO or peroxynitrite results in delayed cell death, wh ich, although occurring by different mechanisms, appears to be mediated by the loss of intracellular redox balance.