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In this study, we investigated the mechanisms of 3′,4′,7-trihydroxyflavone (THF) protection of neuronal cells from neuronal cell death induced by the oxidative stress-related neurotoxin hydrogen peroxide (H₂O₂). Pretreatment with THF significantly elevated cell viability, reduced H₂O₂-induced lactate dehydrogenase (LDH) release, reactive oxygen species (ROS) production, glutathione (GSH) content, superoxide dismutase (SOD) activity, catalase (CAT) activity, and mitochondria membrane potential (MMP) loss. Western blot data demonstrated that THF inhibited the H₂O₂-induced up- or down-regulation of cleaved caspase-3, cleaved caspase-9, cleaved poly-ADP-ribose polymerase (PARP), Bax, Bcl-2, and Bcl-xL, and attenuated the H₂O₂-induced release of cytochrome c from the mitochondria to the cytosol. In addition, pretreatment with THF attenuated H₂O₂-induced rapid and significant phosphorylation of c …