FIGURE 8.

DNA damage participated in H₂O₂-induced neuronal cell Parthanatos. (A) Fluorescence intensity using DCFH-DA showed that H₂O₂ exposure at 250 μmol/L for 24 h significantly induced ROS production in SH-SY5Y cells, which was significantly inhibited by pretreatment with antioxidant NAC. (B) MTT analysis showed that H₂O₂-induced reduction in cell viability of SH-SY5Y cells was markedly reversed with pretreatment of PARP-1 inhibitor 3AB. (C) LDH release assay showed that H₂O₂-induced cell death in SH-SY5Y cells was obviously rescued with pretreatment of PARP-1 inhibitor 3AB. (D–G) Western blotting and quantitative analysis showed that H₂O₂-induced upregulation of cytoplasmic PAR polymer, PARP-1, and AIF both in cytoplasm and nucleus were obviously attenuated by pretreatment of 3AB in SH-SY5Y cells. (H,I) Western blotting and quantitative analysis showed that H₂O₂-induced upregulation of protein 8-OHdG, γH2AX, and p-ATM were obviously attenuated by pretreatment of NAC in SH-SY5Y cells. (J) MTT analysis showed that H₂O₂-induced reduction in cell viability of SH-SY5Y cells was markedly reversed in the presence of NAC. (K) LDH release assay showed that H₂O₂-induced cell death in SH-SY5Y cells was significantly rescued in the presence of NAC. (L–O) Western blotting and quantitative analysis showed that H₂O₂-induced upregulation of cytoplasmic PAR polymer, PARP-1, and AIF both in cytoplasm and nucleus were obviously attenuated by pretreatment of NAC in SH-SY5Y cells. Compared with the control group, ^∗∗p < 0.01; Compared with H₂O₂ group, significant differences were shown in H₂O₂ group pretreated with 3AB or NAC (p < 0.01). Data were presented as mean ± SD from five independent experiments.