Pronuciferine is a naturally occurring proaporphine alkaloid that belongs to isoquinoline alkaloids. The aim of this study is to investigate the neuroactivity of pronuciferine. We assessed the neuroprotective effect of pronuciferine against hydrogen peroxide (H2O2)-induced apoptosis in human neuronal SH-SY5Y cells. In addition, we measured the effect of pronuciferine on cell metabolites and brain-derived neurotrophic factor (BDNF) level in SH-SY5Y cells. In vitro result shows that pronuciferine at 10 mu M significantly (P < .001) increased the proliferation of SH-SY5Y by 45%, and upon H(2)O(2)addition, pronuciferine significantly (P < .001) suppressed neuronal death caused by H2O2. Gas Chromatography-Mass Spectrometry (GC-MS) metabolomics study revealed that pronuciferine has a high impact on glycine-serine-threonine pathway by changing the intracellular level of serine dimethylglycine, sarcosine, and threonine. Also, pronuciferine increased the intercellular level of aspartic acid, glutamine, and tryptophan. Additionally, pronuciferine significantly (P < .05) increased the intracellular BDNF protein expression at 10 mu M. Therefore, pronuciferine is a neuroactive molecule that might act as a neuroprotective agent to prevent apoptosis in neurodegenerative diseases.